Anti-bodies

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- All Roads Lead to Terrain -

ANTIBODIES MISCONEPTION

The Question of REINFECTION

"...the question of immunity is one of the biggest unknowns. Whether infection confers immunity to reinfection “is uncertain,” wrote Newcastle University academics in a paper published in the Journal of Infection in December 2020....

...Out of 6614 participants, 44 had “possible” or “probable” reinfections...

...Worldwide, 31 confirmed cases of covid-19 reinfection have been recorded, although that could be an underestimate....

https://www.bmj.com/content/372/bmj.n99

AHH HAA

De Jong, 22, had tested positive on 17 April and suffered mild symptoms for about 2 weeks. She tested negative on 2 May—just in time to say farewell to her dying grandmother—and returned to work as a nursing intern in a hospital in Rotterdam, the Netherlands.

But when her symptoms re-emerged, her doctor suggested she get tested again. “A reinfection this soon would be peculiar, but not impossible,” she told De Jong, who by then had again lost her sense of smell and had abdominal pains and diarrhea.

https://www.sciencemag.org/news/2020/11/more-people-are-getting-covid-19-twice-suggesting-immunity-wanes-quickly-some?fbclid=IwAR3KQs7c5lBlIem0aQ0ibbZr2kjCrWxZSjCU2_bP8MS69pcx15f6jJJ6BPc

The woman was first diagnosed with Covid-19 in late January, according to a statement released by Osaka’s prefectural government Wednesday. She was discharged shortly after, once her symptoms had improved. A subsequent test came back negative for the virus. Three weeks later she returned with a sore throat and chest pain and tested again.

You may have given blood and been told you have antibodies or had COVID-19 and trusted your body to fight off the infection. However, health officials say there is no hard evidence showing the antibodies built up can protect against reinfection.

someone testing positive for antibodies doesn’t mean they are truly resistant to the disease, and it does not tell us they are resistant to infection,” said Taylor.

(THIS GOES FOR A VACCINE ALSO, TERRAIN THEORY)

https://www.news9.com/story/6008cfce6d58690bbf3a86bf/state-epidemiologist-says-no-clear-evidence-to-show-antibodies-protects-against-covid19-reinfection?fbclid=IwAR1hcOcKzFC3xqu_SYsTgMcLSc-2PSthPlLr3qV2dabu7t7wAPAjbkSQsGs

At least 285 individuals in Mexico appear to have contracted the novel coronavirus twice, according to a preprint posted October 18 on medRxiv. ...

“If we find that our immunity is poor, or nonexistent . . . this will be a big problem for vaccination policies,” study coauthor Carlos Hernandez-Suarez, a researcher at Universidad de Colima in San Sebastian, Mexico....

THEY REDEFINED REINFECTION...... BY PCR, NOT SYMPTOMS!

https://www.the-scientist.com/news-opinion/more-sars-cov-2-reinfections-reported-but-still-a-rare-event-68089?fbclid=IwAR3kwZFYr2jbwx9u89ejvfXCHPP_hAs4MfhMEnRGIto52kejpQOTlsjKR_M

So again confirmation that a positive laboratory result is insignificant. The question arises again and again as to how you know that antibodies offer circulating protection when the highest authorities themselves say that a titer increase cannot prove protection exists. When people have high antibody levels, do they still fall ill? If no one can say exactly at what titer level there is real protection, why is the approval of a vaccine based on that exact reading? Personally, this makes me more than a little suspicious.

https://truthseeker.se/wp-content/uploads/Stefan-Lanka-The-Misinterpretation-of-the-Antibodies-English-Translation.pdf?fbclid=IwAR2JSmRoQzgNtl3ak85sIVj4l8fm7RMhefLHQPL4lXhdNgmZJiOHS2nD1TI

Underlying the birth of virology was the doctrine of monomorphism--that all microorganisms are fixed species, unchangeable; that each pathological type produces only one specific disease; that microforms never arise endogenously, i.e., have absolute origin within the host; and that blood and tissues are sterile under healthy conditions. Theoretically, under ideal health conditions, the blood might be sterile, though it has the inherent potential to develop morbid microforms, as discussed earlier. Long and repeated observation of live blood in the phase-contrast, dark-field microscope, however, shows that the blood can contain various microforms in an otherwise asymptomatic host, or in a condition, or in a condition defined as normal or healthy in orthodox terms. Monomorphism was the cornerstone of developments in 20th-century medical research and treatments. Refusal by the mainstream to examine fairly, much less accept, the demonstrated facts of pleomorphism--that viruses and bacteria, yeast, fungi and mold, are evolutions from microzyma; that microforms can rapidly change their form (evolve and devolve) in vivo, one becoming another, dependent upon conditions in the biological terrain (environment); that blood and tissues are not necessarily sterile; and that there are no specific diseases, but only specific disease conditions--was the foundation of the debate. It is so called because those who wore the "robes" of scientific authority would not be swayed from folly when resented with its contrary proofs. These proofs began in earnest with Antoine Béchamp in the nineteenth-century.

http://www.medicinacomplementar.com.br/.../mb-0464.pdf

It became clear to me durring 2020 when I had spent most of the 12 months learning the problems with "pandemic" narrative, the issues with Virology and finally being introduced to Terrain Model, that reports on "reinfection" of "Covid" (or showing symptoms twice ) was also pointing to Terrain Model. Although Virology being so loosely formed around hypothosis always has some excuse as to why its failing to explain anything and fails to show supporting evidence. Its almost comical to hear Fauci talk about antibodies.

WE HAVE BEEN SOLD ON THIS ASSUMPTION, YES ASSUMPTION THAT:

A piece of a live or dead virus created immunity and taught our body to create antibodies when encountering a similar antigen or pathogen. Here is the history of that great assumption that accopanied the fictious "Virus" Hoax.

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LINUS PAULING'S 1940 ANTIBODY STRUCTURE THEORY:

Linus Pauling was an American Chemist who published a vast library of work with well over 1200 papers and books. He is the only person to win two unshared Nobel Prizes and is considered one of the 20 greatest scientists of all time. Pauling also gets the distinction of "proving" correct Paul Ehrlich's lock and key mechanism for how antibodies/antigens work:

"Ehrlich proposed a ‘lock and key’ mechanism of antibody-antigen interaction but this WAS NOT CONFIRMED UNTIL THE 1940s BY LINUS PAULING."

https://www.rcpath.org/uploads/assets/077f9015-91d1-41a4-ba3fd408b884967b/17-Structure-of-an-Antibody.pdf

"THE NEXT MAJOR ADVANCE WAS IN THE 1940s, WHEN LINUS PAULING CONFIRMED THE LOCK-AND-KEY THEORY PROPOSED BY EHRLICH by showing that the interactions between antibodies and antigens depend more on their shape than their chemical composition."

https://en.m.wikipedia.org/wiki/Antibody

Pauling was able to prove Ehrlich's theoretical explanations and drawings correct by...providing his own theoretical explanations and drawings. And depending on what source you look to, he was either correct in his theory or entirely wrong:

"Pauling was intrigued by Landsteiner’s work, and began reading about antibodies; he was interested and puzzled by what he found. While the scientific community knew that antibodies worked, HOW EXACTLY THEY WORKED AND HOW EXACTLY THEY WERE FORMED WERE STILL UNKNOWN.

At the time, there were FOUR MAIN SCHOOLS OF THOUGHT REGARDING THE CREATION OF ANTIBODIES: the Antigen-Incorporation theory, the Side-Chain theory, the Instruction theory, and the Selection theory."

"The Instruction theory states that the body uses antigens as a template, then manufactures antibodies to specifically combat the antigen that the antibody is based off of. PAULING EVENTUALLY BELONGED TO THIS SCHOOL OF THOUGHT, as did Landsteiner, Michael Heidelberger, Felix Haurowitz, and Jerome Alexander. THIS GROUP WAS FAR FROM UNIFIED HOWEVER; the only point on which adherents to this school agreed was that antigens acted as templates. HOW ANTIBODIES WORKED, AND HOW THEY WERE PRODUCED, WAS STILL A HIGHLY CONTENTIOUS QUESTION."

"Another argument developed in the 1940 paper was that antibodies are bivalent – that is, they have two sites which can bind to antigens. In addition to being bivalent, PAULING HYPOTHESIZED THAT EACH OF THE “ARMS” OF AN ANTIBODY COULD LATCH ONTO DIFFERENT KINDS OF ANTIGENS. While PAULING WAS INCORRECT ON THE LATTER PART – antibodies can only grab onto one type of antigen – he was correct that they are bivalent.

Pauling had gotten off to a strong and noticeable start in the field of immunology. WHETHER CORRECT OR INCORRECT, he was making progress towards a greater understanding of how the body protects itself."

https://paulingblog.wordpress.com/2013/08/01/thinking-about-the-creation-of-antibodies/

"THE MODEL FIT A GREAT DEAL OF DATA. It was simple and clean. Pauling wrote it up and SENT WHAT HE HOPED WOULD BE A LANDMARK PAPER ON ANTIBODY FORMATION to the Journal of the American Chemical Society in the summer of 1940. "What is the simplest structure which can be suggested . . . for a molecule with the properties observed for antibodies, and what is the simplest reasonable process of formation for such a molecule?" he asked at the beginning. Then he elegantly and persuasively put forward his ideas.

HIS PAPER MADE A GREAT STIR. It was another demonstration of the power of applying chemical ideas to biology. It was a new vindication of Weaver's approach, which he was now calling "molecular biology."

IT WAS ONLY LATER THAT IT WAS PROVEN COMPLETELY WRONG."

http://scarc.library.oregonstate.edu/.../narr.../page17.html

Below are some highlights from Pauling's 15-page Theory:

A THEORY OF THE STRUCTURE AND PROCESS OF FORMATION OF ANTIBODIES

"I. Introduction

During the past four years I have been making

an effort to understand and interpret serological phenomena in terms of molecular structure and molecular interactions. The field of immunology is so extensive and the experimental observations are so complex (AND OCCASIONALLY CONTRADICTORY) that NO ONE HAS FOUND IT POSSIBLE TO INDUCE A THEORY OF THE STRUCTURE OF ANTIBODIES FROM THE OBSERVATIONAL MATERIAL. As an alternative method of attack we may propound and attempt to answer the following questions: WHAT IS THE SIMPLEST STRUCTURE WHICH CAN BE SUGGESTED, on the basis of the extensive information now available about intramolecular and intermolecular forces, for a molecule with the properties observed for antibodies, AND WHAT IS THE SIMPLEST REASONABLE PROCESS OF FORMATION OF SUCH A MOLECULE? Proceeding in this way, I HAVE DEVELOPED A DETAILED THEORY OF THE STRUCTURE AND PROCESS OF FORMATION OF ANTIBODIES and the nature of serological reactions which is more definite and more widely applicable than earlier theories, and which is compatible with our present knowledge of the structure and

properties of simple molecules as well as with

most of the direct empirical information about

antibodies. This theory is described and dis-

cussed below.

11. The Proposed Theory of the Structure and

Process of Formation of Antibodies

When an antigen is injected into an animal

some of its molecules are captured and held in the region of antibody production.2 An antibody to this antigen is a molecule with a configuration which is complementary to that of a portion of the antigen molecule. This complementariness gives rise to specific forces of appreciable strength between the antibody molecule and the antigen molecule; we may describe this as a bond between the two molecules. I ASSUME, with Marrack, Heidelberger, and other investigators, THAT THE PRECIPITATE OBTAINED IN THE PRECIPITIN REACTION IS A FRAMEWORK, AND THAT TO BE EFFECTIVE IN FORMING THE FRAMEWORK AN ANTIBODY MOLECULE

MUST HAVE TWO OR MORE DISTINCT REGIONS WITH SURFACE CONFIGURATION COMPLEMENTARY TO THAT OF THE ANTIGEN. THE RULE OF PARSIMONY (the use of the minimum effort to achieve the result) suggests that there are only two such regions, that is, that the antibody molecules are at the most bivalent. THE PROPOSED THEORY IS BASED ON THIS REASONABLE ASSUMPTION. It would, of course, be possible to expand the theory in such a way as to provide a mechanism for the formation of antibody molecules with valence higher than two ; but this would make the theory considerably more complex, and it is likely that antibodies with valence higher than two occur only rarely, if at all."

"The effect of an antigen in determining the

structure of an antibody molecule might involve the ordering of the amino-acid residues in the polypeptide chains in a way different from that in the normal globulin, as suggested by Breinl and Haurowitz" and Mudd. I ASSUME, HOWEVER, THAT THIS IS NOT SO, BUT THAT ALL ANTIBODY MOLECULES CONTAIN THE SAME POLYPEPTIDE CHAINS AS NORMAL GLOBULIN, AND DIFFER FROM NORMAL GLOBULIN ONLY IN THE CONFIGURATION OF THE CHAIN; that is, in the way that the chain is coiled in the molecule. THERE IS AT PRESENT NO DIRECT EVIDENCE SUPPORTING THIS ASSUMPTION. The assumption is made because, although I HAVE FOUND IT IMPOSSIBLE TO FORMULATE IN DETAIL A REASONABLE MECHANISM WHEREBY THE ORDER OF AMINO-ACID RESIDUES IN THE CHAIN WOULD BE DETERMINED BY THE ANTIGEN, a simple and reasonable mechanism, described below, can be advanced whereby the antigen causes the polypeptide chain to assume a configuration complementary to the antigen. The number of configurations accessible to the polypeptide chain is so great as to provide an explanation of the ability of an animal to form antibodies with considerable specificity for an apparently unlimited number of different antigens,8 without the necessity of invoking also a variation in the amino-acid composition or amino-acid order.

The Postulated Process of Formation of Antibodies.

LET US ASSUME THAT THE GLOBULIN MOLECULE CONSISTS OF A SINGLE POLYPEPTIDE CHAIN, containing several hundred amino-acid residues, and that the order of amino-acid residues is such that for the center of the chain one of the accessible configurations is much more stable than any other, whereas the two end parts of the chain are of such a nature that there exist for them many configurations with nearly the same energy. (This point is discussed in detail in Section IV.) Four steps in our postulated process of formation of a normal globulin molecule are illustrated on the left side of Fig. 1."

"III. Some Points Of with Experiment

a. The Heterogeneity Of Immune Sera

THE THEORY REQUIRES THAT THE SERUM HOMOLOGOUS TO A GIVEN ANTIGEN BE NOT HOMOGENEOUS, BUT HETEROGENEOUS, CONTAINING ANTIBODY MOLECULES OF GREATLY VARIED CONFIGURATIONS. Many of the antibody molecules will be bivalent, with two active ends with configuration complementary to portions of the surface of an antigen molecule. Great variety in this complementary configuration would be expected to result from the accidental approximation to one or another surface region, and further variety from variation in position of the antigen molecule relative to the point of liberation of the globulin chain end and from accidental coiling and linking of the chain end before it comes under the influence of the antigen. Some of the antibody molecules would be univalent, one of the chain ends having, because of its too great distance from the antigen, folded into a normal globulin configuration."

"b. The Bivalence of Antibodies and the

Multivalence of Antigens.

OUR THEORY IS BASED ON THE IDEA THAT THE PRECIPITATE FORMED IN THE PRECIPITIN REACTION IS A NETWORK OF ANTIBODY AND ANTIGEN MOLECULES IN WHICH MANY OR ALL OF THE ANTIBODY MOLECULES GRASP TWO ANTIGEN MOLECULES APIECE AND THE ANTIGEN MOLECULES ARE GRASPED BY SEVERAL ANTIBODY MOLECULES. The direct experimental evidence for this picture of the precipitate has been ably discussed by its propounders and supporters, Marrack and Heidelberger and Kendall, and need not be reviewed here. TO THE STRUCTURAL CHEMIST IT IS CLEAR THAT THIS PICTURE OF THE PRECIPITATE MUST BE CORRECT. The great specificity of antibody-antigen interactions requires that a definite bond be formed between an antibody molecule and an antigen molecule. If antibodies or antigens were univalent, this would lead to complexes of one antigen molecule and one or more antibody molecules (or of one antibody molecule

and one or more antigen molecules), and we know from experience with proteins that these aggregates would in general remain in solution. If both antibody and antigen are multivalent, however, the complex will grow to an aggregate of indefinite size, which is the precipitate."

"IT SEEMS PROBABLE THAT ALL ANTIBODIES HAVE THIS STRUCTURE-THAT THEY ARE BIVALENT, WITH THEIR TWO ACTIVE REGIONS OPPOSITELY DIRECTED. Heidelberger and his collaborators and Marrack have emphasized the multivalence of antibodies and antigens,14 but limitation of the valence of antibodies to the maximum value two (ignoring the exceptional case of the attachment of two or more antigens or haptens to the same end region of an antibody) has not previously been made."

"c. The Antibody-Antigen Molecular Ratio in

Precipitates.

OUR THEORY PROVIDES AN IMMEDIATE SIMPLE EXPLANATION OF THE OBSERVED ANTIBODY-ANTIGEN MOLECULAR RATIOS IN PRECIPITATES. Under OPTIMUM CONDITIONS a precipitate will be formed in which all the valences of the antibody and antigen molecules are satisfied. AN IDEALIZED REPRESENTATION OF A PORTION OF SUCH A PRECIPITATE IS GIVEN IN Fig. 3. The figure shows a part of a layer with each antigen molecule bonded to six surrounding antibody molecules ; this structure represents the value N = 12 for the valence of the antigen, each antigen molecule being attached also to three antibody molecules above the layer represented and to three below. Each antibody

molecule is bonded to two antigen molecules, one at each end. An ideal structure of the antibody-antigen precipitate for N = 12 may be described as having antigen molecules at the positions corresponding to closest packing, with the twelve antibody molecules which surround each antigen molecule lying along the lines connecting it with the twelve nearest antigen neighbors.

SIMILAR IDEAL STRUCTURES CAN BE SUGGESTED FOR OTHER VALUES OF THE ANTIGEN VALENCE. The antigen molecules might be arranged for N = 8 at the points of a body-centered cubic lattice, and for N = 6 at the points of a simple cubic lattice, with antibody molecules along the connecting lines. For N = 4 the antigen molecules, connected by antibody molecules, might lie at the points occupied by carbon atoms in diamond; or two such frameworks might interpenetrate, as in the cuprous oxide arrangement (copper and oxygen atoms being replaced by antibody and antigen molecules, respectively).

IT IS NOT TO BE INFERRED THAT THE ACTUAL PRECIPITATES HAVE THE REGULARITY OF STRUCTURE OF THESE IDEAL ARRANGEMENTS. The nature of the process of antibody formation, INVOLVING THE USE OF A PORTION OF THE ANTIGEN SURFACE SELECTED AT RANDOM AS THE TEMPLATE FOR THE MOLDING OF AN ACTIVE END OF AN ANTIBODY MOLECULE, INTRODUCES SO MUCH IRREGULARITY IN THE FRAMEWORK THAT A REGULAR STRUCTURE ANALOGOUS TO THAT OF A CRYSTAL IS PROBABLY NEVER FORMED. The precipitate is to be compared rather with a glass such as silica glass, in which each silicon atom is surrounded tetrahedrally by four oxygen atoms and each oxygen atom is bonded to two silicon atoms, but which lacks further orderliness of arrangement. Additional disorder is introduced in the precipitate by variation in the effective valence of the antigen molecules and by the inclusion of antibody molecules with only one active end."

"IT IS SEEN THAT OUR THEORY PROVIDES A SIMPLE EXPLANATION OF THE FACT THAT FOR ANTIGENS OF MOLECULAR WEIGHT EQUAL TO OR LESS THAN THAT OF THE ANTIBODY THE PRECIPITATE CONTAINS CONSIDERABLY MORE ANTIBODY THAN ANTIGEN. The values given in Table I are not to be considered as having rigorous quantitative significance. The calculated maximum molecular ratio would be larger for elongated antibody molecules than for spherical antibody molecules, and larger for non-spherical than for spherical antigen molecules, and, moreover, in

many sera antibodies might be complementary in the main only to certain surface regions of the antigen, the number of these determining the valence of the antigen. That this is so is indicated by the observation that after long immunization of a rabbit with egg albumin serum was obtained giving a precipitate with a considerably larger molecular ratio than that for earlier bleedings."

"d. The Use of a Single Antigen Molecule

as the Template for an Antibody Molecule.

THERE ARE TWO WAYS IN WHICH AN ANTIBODY MOLECULE WITH TWO OPPOSED ACTIVE REGIONS COMPLEMENTARY TO THE ANTIGEN MIGHT BE PRODUCED. One is the way described in Section 11. The other would involve the which (with an occasional exception) both antigen manufacture of the antibody molecule in its and antibody are bivalent, the molecular ratio final configuration between two antigen molecules, one of which would serve as the pattern for one antibody end and the other for the second. NO ATTEMPTS TO DECIDE BETWEEN THESE ALTERNATIVES SEEMS TO HAVE BEEN MADE BEFORE; evidence, however, some of which is mentioned below, to indicate that the first method of antibody production, involving only one antigen molecule, occurs predominantly. IT IS FOR THIS REASON THAT I HAVE DEVELOPED THE RATHER COMPLICATED THEORY DESCRIBED ABOVE, with the two end portions of the antibody forming first, one (or both) then separating from the antigen, and the central part of the antibody then assuming its shape and holding the active ends in position for attachment to two antigen molecules.

THIS THEORY REQUIRES THAT THE FORMATION OF ANTIBODY BE A REACTION OF THE FIRST ORDER WITH RESPECT TO THE ANTIGEN, whereas the other alternative would require it to be of the second order. THERE EXISTS VERY LITTLE EVIDENCE AS TO WHETHER ON IMMUNIZATION WITH SMALL AMOUNTS OF ANTIGEN THE ANTIBODY PRODUCTION IS PROPORTIONAL TO THE AMOUNT OF ANTIGEN INJECTED OR TO ITS SQUARE."

"e. Criteria for Antigenic Power.

THERE HAS BEEN EXTENSIVE DISCUSSION OF THE QUESTION OF WHAT MAKES A SUBSTANCE AN ANTIGEN, BUT NO GENERALLY ACCEPTED CONCLUSIONS HAVE BEEN REACHED. OUR THEORY PERMITS THE FORMULATION OF THE FOLLOWING REASONABLE CRITERIA FOR ANTIGENIC ACTIVITY:

1. The antigen molecule must contain active groups, capable of sufficiently strong interaction with the globulin chain to influence its configuration.

2. The configuration of the antigen molecule

must be well-defined over surface regions large enough to give rise to an integrated antibody-

antigen force sufficient to hold the molecules

together.

3. The antigen molecule must be large enough

to have two or more such surface regions, and in case that the antigenic activity depends upon a particular group the molecule must contain at least two of these groups. (This criterion applies to antibodies effective in the precipitin and agglutinin reactions and in anaphylaxis.)"

"IV. A More Detailed Discussion of the Struc-

ture of Antibodies and Other Proteins

THERE HAS BEEN GATHERED SO FAR VERY LITTLE DIRECT EVIDENCE REGARDING THE DETAILED STRUCTURE OF PROTEIN MOLECULES. Chemical information is compatible with the polypeptide-chain theory of protein structure, and this theory is also supported by the rather small amount of pertinent X-ray evidence.25"

"LAYER STRUCTURES OTHER THAN THIS ONE MIGHT ALSO BE ASSUMED, in which the chains are not extended. Some fibrous proteins, such as a-keratin, are known to have structures of this general type, BUT THE NATURE OF THE FOLDING OF THE CHAINS HAS NOT YET BEEN DETERMINED.

WE HAVE POSTULATED THE EXISTENCE OF AN EXTREMELY LARGE NUMBER OF ACCESSIBLE CONFIGURATIONS with nearly the same energy for the end parts of the globulin polypeptide chain. A layer structure, with variety in the type of folding in the layer, would not, it seems to me, give enough configurational possibilities to explain the great observed versatility of the antibody precursor in adjusting itself to the antigen, and I think that skew configurations must be invoked.``

V. Further Comparison of the Theory with

Experiment. Possible Experimental Tests of

Predictions

a. Methods of Determining the Valence of

Antibodies.

"The bivalence of antibodies and our postulate that only one antigen molecule is involved in the formation of an antibody molecule require that a precipitin-effective antihapten be produced only if the injected antigen contains at least two hapten groups. PERTINENT DATA HAVE BEEN OBTAINED ON THIS POINT BY HAUROWITZ and his collaborators,"

FROM THE FOOTNOTES:

"From the data discussed above, WHICH IN OUR OPINION indicate that two hapten groups combine with a bivalent antibody molecule,

HAUROWITZ DREW THE DIFFERENT CONCLUSIONS that one arsenic-containing group in the antigen combines with one antibody molecule."

"IF WE ASSUME THAT THE CONDITIONS OF EACH PRECIPITATION WERE SUCH THAT ONLY SUITABLE BIVALENT ANTIBODY MOLECULES WERE INCORPORATED IN THE PRECIPITATE, the equations above would require the third precipitate to weigh about 24 mg. ; the experiment accordingly provides some support for the theory."

"THE QUALITATIVE EXPERIMENTAL RESULTS which have been reported ARE IN PART COMPATIBLE AND IN PART INCOMPATIBLE WITH THE THEORY."

VI. Processes Auxiliary to Antibody Formation

"It seems not unlikely that certain processes

auxiliary to antibody formation occur. The reported increase in globulin (aside from the antibody fraction) after immunization suggests the operation of a mechanism whereby the presence of antigen molecules accelerates the synthesis of the globulin polypeptide chains. THERE IS LITTLE BASIS FOR SUGGESTING POSSIBLE MECHANISMS FOR THIS PROCESS AT PRESENT."

"Summary

IT IS ASSUMED THAT ANTIBODIES DIFFER FROM NORMAL SERUM GLOBULIN ONLY IN THE WAY IN WHICH THE TWO END PARTS OF THE GLOBULIN POLYPEPTIDE CHAIN ARE COILED, these parts, as a result of their amino-acid composition and order, having accessible a very great many configurations with nearly the same stability; under the influence of an antigen molecule they assume configurations complementary to surface regions of the antigen, thus forming two active ends. After the freeing of one end and the liberation of the central part of the chain this part of the chain folds up to form the central part of the antibody molecule, with two oppositely-directed ends able to attach themselves to two antigen molecules.

Among the points of comparison of the theory

and experiment are the following: the heterogeneity of sera, the bivalence of antibodies and multivalence of antigens, the framework structure and molecular ratio of antibody-antigen precipitates, the use of a single antigen molecule as template for an antibody molecule, criteria for antigenic activity, the behavior of antigens containing two different haptens, the antigenic activity of antibodies, factors affecting the rate of antibody production and the specificity of anti-

bodies, and the effect of denaturing agents. It

is shown that MOST OF THE REPORTED EXPERIMENTAL RESULTS ARE COMPATIBLE WITH THE THEORY. Some new experiments suggested by the theory are mentioned."

https://doi.org/10.1021/ja01867a018

In Summary:

-the science and findings of Immunology were so vast, complex, and contradictory that no one had attempted to describe the structure and formation of antibodies

-Pauling based his theories on what could be the simplest explanation for the structure and formation of an antibody

-he ASSUMED that the precipitate in the precipitin reactions was the framework for an antibody and that there must be 2 or more regions complementary to an antigen

-he based his assumptions of the form of an antibody on the rule of parsimony (the use of minimum effort to achieve a result)

-he ASSUMED that all antibodies are similar to normal globulins and differ only in the configuration of their chains

-however. he admits there is no direct evidence supporting his assumptions

-Pauling based his assumption on the fact that he could not formulate a reasonable mechanism where the amino-acid chains were determined by the antigen

-he once again ASSUMES that the globulin molecule is a single polypeptide chain containing several hundred amino-acid residues

-his theory requires that the serum homogulous to the given antigen not be homogeneous but heterogeneous containing antibody molecules consisting of many configurations

-his theory is based on the idea that the precipitate is a network of antibody/antigen molecules in which antibody molecules grasp two antigen molecules a piece and the antigens are grasped by several antibody molecules

-he states the picture of the precipitate is clear to structural chemists so it must be correct

-he states it is PROBABLE that antibodies are bivalent

-he believed his theory provided a simple explanation for the antibody-antigen molecular ratios in precipitates under optimum conditions

-Pauling provided drawings of idealized representations of antibodies and antigens

-however, he states it should not be inferred that the precipitates have the regularity of structure as presented in the idealized versions

-he states the nature of antibody formation, using a portion of the antigen surface selected at random as the template for the molding of an active end of an antibody molecule, probably never forms the perfect crystalline structure represented

-Pauling states there are two ways in which antibodies with two-opposing active regions complementary to the antigen might be produced, yet no attempts have been made to decide between them, which is why he created his complicated theory

-his theory requires that the formation of the antibody be a reaction of the first order in regards to the antigen

-Pauling says there have been extensive discussions of what makes a substance an antigen but accepted conclusions have not been reached

-he states his theory allows for the formulation of reasonable criteria for antigenic activity

-he admits there has been very little direct evidence on the detailed structure of protein molecules

-there are potential layer structures which can be assumed but no determination has been made on the folding of the chains

-he postulated the existence of very large numbers of accessible configurations

-Pauling provides evidence from Haurowitz which he states supports his conclusion that two hapten groups combine with the bivalent antibody but admits in the footnotes that Haurowitz came to a different conclusion

-he ASSUMES that the conditions in the precipitation were such that only suitable bivalent antibody molecules were present in the precipitate

-he admits experimental results are somewhat compatible and also somewhat incompatible with his theory

-he admits there is little evidence to propose mechanisms for how globulin increases after immunization

-Pauling concludes by stating it is ASSUMED that antibodies differ from serum globulin only in the way the two globulin polypeptide chains are coiled

-he states most of the experimental evidence is compatible with his theory

If you enjoy your "proof" in the form of assumptions and guesswork based on cherry-picked experimental research, Pauling's paper will be right up your alley. The amount of times he admits to making assumptions is astonishing. Granted, it is admitted to be a THEORY on the structure and formation of antibodies. However, it is lauded by some as proof of the form of antibodies as well as confirmation of Ehrlich's own theory regarding the lock-and-key mechanism of action. One unproven theoretical explanation does not get to confirm another one.

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PAUL EHRLICH'S 1900 SIDE-CHAIN THEORY PART 2: THE COMPLEMENT SYSTEM

The last part of Ehrlich's presentation to the Royal Society formed around the idea of the complement system. In short, this is a part of the immune system that complements the ability of antibodies to clear out the damaged cells and the inhabiting microbes through inflammation in order to attack the pathogen and eliminate it from the body. A brief overview of Ehrlich's role in the creation of this theory:

"Paul Ehrlich described the side-chain theory of antibody formation, especially the mechanisms of antibody neutralisation by toxins that induced bacterial lysis with the help of complement (which has replaced the historical term alexin). According to his theory, the immune cells contained receptors that could recognise antigens, and following immunisation, these receptors multiplied and were shed into the circulation as ‘amboceptors’ (NOW CALLED ANTIBODIES). These antibodies attached not only to specific antigens but also to a heat-labile antimicrobial component called ‘complement’ [8, 9]. EHRLICH'S THEORY PROPOSED THAT THE ANTIBODY AND COMPLEMENT COMBINED TO FORM A COMPLEX ENZYME CAPABLE OF ATTACKING AND KILLING CELLS AND MICROORGANISMS. In the ensuing years, this concept had a protagonist in the form of Bordet who argued that the antigen-antibody union was reversible, CONTRADICTING EHRLICH'S VIEW THAT THE ANTIGEN-ANTIBODY UNION WAS A FIRM AND BASED ON STEREO CHEMICAL SPECIFICITY [10]. Ehrlich’s concept EMPHASIZED THE PRESENCE OF MULTIPLE ANTIGENS AND COMPLEMENTS IN THE SERUM, while Bordet’s view revolved around a ‘single complement’ component that bound non-specifically to the antigen."

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3956958/

Below are some highlights from the remainder of Ehrlich's report on his theoretical investigation into immunity which details the complement system. I provided some additional insight afterwards regarding how this theory was utilized:

"Bordet showed shortly thereafter, that in the case quoted there was present in the serum a specific hsemolysine which dissolved the blood corpuscles of the rabbit. He also proved that these hsemolysines as had already been shown by Buchner and Daremberg in the case of similarly acting bodies which are present in normal blood lost their solvent property on being maintained during half an hour at a temperature of 55° C. Bordet added, further, the new fact, that the blood-solvent property of these sera which had been deprived of solvent

power by heat, the solvent action could be restored if certain normal sera were added to them.

By this important observation an exact analogy was established with the facts of bacteriolysis as elicited by the work of Pfeiffer, Metchni-

koff, and Bordet. In the work on the Pfeiffer phenomenon of bacteriolysis, it had already been ascertained that the solution of bacteria

by specific bacteriolysines WAS BROUGHT ABOUT BY THE COMBINED ACTION OF TWO DIFFERENT BODIES: ONE WHICH WAS SPECIFIC, AROSE DURING THE IMMUNISATION AND WAS STABLE; AND ANOTHER, A VERY UNSTABLE BODY, WHICH WAS PRESENT IN NORMAL SERUM.

In collaboration with Dr. Morgenroth, I have sought in regard to this question, for which haemolysis offered prospects favourable to

experimentation, to make clear the mechanism concerned in the action of these two components—THE STABLE, WHICH MAY BE DESIGNATED “IMMUNE BODY,” AND THE UNSTABLE, WHICH MAY BE DESIGNATED “COMPLEMENT”—which,' acting together, effect the solution of the red blood corpuscles. For this purpose, in the first place, solutions containing either only the “ immune body ” or only the “ complement ” were brought in contact with suitable blood corpuscles, and after separation of the fluid and the corpuscles by centrifugalising, we investigated whether these substances had been taken up by the red blood corpuscles or remained behind in the fluid. The proof of its location in the one position or in the other was readily forthcoming, since to restore to the hsemolysine its former activity, IT WAS ONLY NECESSARY TO ADD TO THE “IMMUNE BODY” A FRESH SUPPLY OF “COMPLEMENT,” OR TO THE “COMPLEMENT” A FRESH SUPPLY OF “IMMUNE BODY,” in order that the presence of the hsemolysine in its integrity might be shown by the occurrence of solution of the blood-cells.

The experiments proved that, after centrifugalising, THE “IMMUNE BODY” IS QUANTITATIVELY BOUND TO THE RED BLOOD CORPUSCLES, AND THAT THE “COMPLEMENT,” ON THE CONTRARY, REMAINS ENTIRELY BEHIND IN THE FLUID.

The presence of the two components in contact with blood corpuscles only occasions the solution of these at higher temperatures, and not at 0° C. And an active haemolytic serum (with “immune body” and "complement” both present) having been placed in contact with red blood corpuscles and maintained for a while at 0° C., it was found after centrifugalising that, under these circumstances also, the “immune body” had united with the red blood corpuscles, but that the “complement” remained in the serum. This experiment showed that both components must, at a temperature of 0° C., have existed alongside one another in a free condition.

But when analogous experiments were undertaken at a higher temperature it was found that both components were retained in the sediment.

THESE FACTS CAN ONLY BE EXPLAINED BY MAKING CERTAIN ASSUMPTIONS REGARDING THE CONSTITUTION OF THE TWO COMPONENTS, i.e., OF THE “IMMUNE BODY” AND THE “COMPLEMENT.” In the first place, two haptophore groups must be ascribed to the “immune body,” one having a great affinity for a corresponding haptophore group of the red blood corpuscles and with which at lower temperatures it quickly unites, and another

haptophore group of a lesser chemical affinity, which at a higher temperature becomes united with the “complement” present in the

serum. Therefore, at the higher temperature, the red blood corpuscles will draw to themselves those molecules of the “immune body” which in the fluid have previously become united with the “complement.”

In this case the “immune body” represents in a measure the connecting chain which binds the complement to the red blood corpuscles, and

so brings them under its deleterious influence. Since under the influence of the “complement”—at least, in the case of the bacteria— appearances are to be observed (for example, in the Pfeiffer phenomenon) which must be regarded as analogous to digestion, we shall not seriously err if we ascribe to this “complement” a ferment-like character.

IT IS OBVIOUS THAT WHEN THE NORMAL SERUM OF ONE ANIMAL POSSESSES HAEMOLYTIC ACTION ON THE BLOOD OF ANOTHER, the component of the hsemolysine which here unites with the red blood corpuscle and forms the connecting link between it and the “complement” which is essential to the occurrence of solution, CANNOT, IN THE ABSENCE OF ANY PRECEDING PROCESS OF IMMUNISATION, BE DESIGNATED “IMMUNE BODY.” In its characteristics and action, however, IT ONLY DIFFERS FROM THIS IN OCCURRING NATURALLY, AND MAY WELL BE DESIGNATED “INTERMEDIATE BODY”

(Zwischenkorper). It may here be stated that the constitution of a haemolysine is graphically represented in fig. 7, Plate 7.

Very important for the conclusion that only with the assistance of the “intermediate body” or of the “immune body” can the “comple-

ment,” which leads to the solution, become united with the blood corpuscle, is the following experiment. The serum of the dog has very considerable solvent action upon guinea-pig’s blood, but loses this property if warmed. If dog’s serum, thus rendered inactive by warming, is brought into contact with suspended corpuscles of guinea-pig’s blood,

these are not dissolved; but, if to such a mixture there are also added guinea-pig serum, i.e., the serum normal to these red blood corpuscles, the erythrocytes are at once dissolved. HERE THE ONLY EXPLANATION IS THAT THE “INTERMEDIATE BODY,” which possesses a specific affinity for guinea-pig erythrocytes, and is present in the inactive dog’s serum, IS ABLE TO SEIZE ON ONE OF THE MANY “COMPLEMENTS” PRESENT IN GUINEA-PIG’S SERUM, with the result that the “complement” which cannot normally attach itself to the corpuscles, comes now to exercise its destructive influence.

We see at the same time from this experiment that the HSEMOLYSINES OCCURRING NATURALLY, OBEY THE SAME LAWS AS THOSE PRODUCED THROUGH THE PROCESS OF IMMUNISING. In fact, for them also, in a great number of instances, precisely similar behaviour has been demonstrated.

The character of the specific union made it possible to find solutions for a number of important questions. In the first place, regarding the multiplicity of the heemolysines, which occur normally in serum, it is well known that numerous sera are able to dissolve blood corpuscles of different species. For example, serum of the dog dissolves blood corpuscles of the rabbit, guinea-pig, rat, goat, sheep, &c. The complex nature of these haemolysines has been already indicated.

Another question arises whether in a serum that is capable of such manifold action there is present one single haemolysine that destroys different red blood-cells, or whether a whole series of hsemolysines come into action, of which one is adapted to guinea-pig blood, another to rabbit blood, &c. The solution of this question may be approached in another way. The serum may be rendered inactive by heat, and then placed in contact with red blood corpuscles of a given kind. Then, supposing, for example, that rabbit blood has been employed, it is found that if the fluid is freed from the erythrocytes by centrifugalisation and the “complement” afterwards added, it is no longer in a position to dissolve rabbit blood, but has not suffered any impairment of its action on other kinds.

By this method of elective absorption it is proved that the normally occurring hsemolysines which chain the blood corpuscles of the rabbit to themselves, are specifically adapted to this purpose. IF WITH

SUITABLE ADJUSTMENT OF CONDITIONS SIMILAR EXPERIMENTS BE CONDUCTED WITH OTHER KINDS OF BLOOD, RESULTS ARE OBTAINED WHICH FORCE US TO THE CONVICTION THAT IN SUCH A SERUM ACTING ON VARIOUS KINDS OF BLOOD THERE ARE PRESENT ABSOLUTELY DIFFERENT “INTERMEDIATE BODIES” (analogues of the “immune bodies”), OF WHICH EACH ONE IS SPECIFIC FOR ONE KIND OF BLOOD, i.e., one is adapted for rabbit’s blood, a second for calf’s blood, &c. Dr. Morgenroth and I have in some cases, indeed, SUCCEEDED IN PROVING THAT THE “COMPLEMENTS” WHICH ARE ADAPTED TO FIT THEMSELVES TO THESE “INTERMEDIATE BODIES,” AND OCCUR IN NORMAL SERA, DIFFER AMONG THEMSELVES. If we reflect that in normal blood, in addition to these different hsemolysines, there are besides a long series of analogous bodies, agglutinines of very different kinds, bacteriolysines, enzymes, anti-enzymes, WE ARE BROUGHT MORE AND MORE TO THE CONVICTION THAT THE BLOOD SERUM IS THE CARRIER OF SUBSTANCES INNUMERABLE AS YET LITTLE KNOWN OR CONCEIVED OF.

Having obtained a precise conception of the method of action of the lysines of the serum—of the hsemolysines, and thereby also of the

bacteriolysines—it becomes possible for us to attempt to solve the mystery of the origin of these bodies. I have in the beginning of this lecture fully developed the “side-chain theory,” according to which the antitoxines are merely certain of the protoplasm “side-chains,”

which have been produced in excess and pushed off into the blood.

The toxines, as secretion products of cells, are in all likelihood still relatively uncomplicated bodies; at least, by comparison with the primary arid complex albumins of which the living cell is composed. If a cell of the organism has, with the assistance of an appropriate “side-chain,” fixed to itself a giant molecule, as the proteid molecule really is, then, with the fixation of this molecule, there, is provided one of the conditions essential for the cell nourishment. Such giant molecules cannot at first be utilised by the cells, and are only made available when, by means of a ferment-like process, they are split into smaller fragments. THIS WILL BE VERY EFFECTUALLY ATTAINED IF, FIGURATIVELY SPEAKING, THE “TENTACLE” OR GRAPPLING ARM OF THE PROTOPLASM POSSESSES A SECOND HAPTOPHORE GROUP ADAPTED TO TAKE TO ITSELF FERMENT-LIKE MATERIAL OUT OF THE BLOOD FLUID. Through such complex organisation, by which the “ tentacle” acts also as the bearer of a ferment-functioning group, this group is brought into close relation with the prey destined to be digested and assimilated.

For such appropriate arrangements, in which THE “TENTACULAR” APPARATUS ALSO EXERCISES A DIGESTIVE FUNCTION—IF IT BE PERMISSIBLE TO PASS FROM THE ABSTRACT TO THE CONCRETE—we find analogies in the different forms of insectivorous plants. Thus it has been known since the famous researches of Darwin that the tentacles of Drosera secrete a proteid-digesting fluid.

If we now recognise that the different lysines only arise through absorption of highly complex cell material—such as red blood corpuscles or bacteria—then the explanation, in accordance with what I have said, is that there are present in the organism “side-chains”

of a special nature, so constituted that they are endowed not only with an atomic group by virtue of the affinities of which they are

enabled to pick up material, but also with a second atomic group, which, being ferment-loving in its nature, brings about the digestion of the material taken up. SHOULD THE PUSHING-OFF OF THESE “SIDE-CHAINS” BE FORCED, AS IT WERE, BY IMMUNISATION, THEN THE "SIDE-CHAINS" THUS SET FREE MUST POSSESS BOTH GROUPS, AND WILL THEREFORE IN THEIR CHARACTERISTICS ENTIRELY CORRESPOND TO WHAT WE HAVE PLACED BEYOND DOUBT AS REGARDS THE “IMMUNE-BODY” OF THE HSEMOLYSINE.

In this manner is simply and naturally explained the astonishingly specialised arrangement that, through the introduction of a definite bacterium into the body, SOMETHING IS PRODUCED WHICH IS ENDOWED WITH THE POWER OF DESTROYING BY SOLUTION THE BACTERIUM WHICH WAS ADMINISTERED AND NO OTHER. This contrivance of the organism is to be regarded as nothing more than a repetition of a process of normal cell-life, and the outcome of primitive wisdom on the part of the protoplasm."

https://doi.org/10.1098/rspl.1899.0121

In Summary:

-Ehrlich states that there are two different bodies acting in combination for bacteriolysines (the rupture of a bacterial cell by antibodies): a stable one brought about by immunization and an unstable one already present in the blood

-he called the stable element the "immune body" and the unstable element the "complement"

-after centrifugation, he states the "immune body" stays with the red blood cells while the "complement" is left behind

-through his experiments, he determined that temperature had an effect on whether or not the substances were left behind after centrifugation

-in order to explain these facts, Ehrlich states that ASSUMPTIONS about the two bodies need to be made

-he states that when the blood of a non-immunized animal has haemolytic action on the blood of another animal, this can not be considered the action of the "immune body"

-he termed this phenomena the "intermediate body" even though it had exactly the same action and characteristics of the "immune body" and the only difference was it occurred naturally

-he states that the process of hsemolysines (lysis of the red blood cells) is the same when it occurs naturally or through immunizations

-he states that if experiments are carried out on various blood types, and there are multiple "intermediate bodies" specific for each kind of blood (rabbits blood, calf blood, Guinea pig blood, etc.), then he succeeded in proving

that there are different complements that fit these "intermediate bodies"

-he was convinced the blood contains innumerable substances for which little is known or conceived of

-he talks of a "tentacle" aiding in the process of digestion/fermentation to eliminate toxins

-he says his abstract "tentacular" apparatus is part of the digestive function

-he believes immunization forces the pushing off of "side-chains" (antibodies) into the bloodstream

This process of binding serum complement was yet another indirect method used in an attempt to prove antibodies exist. If a reaction occurs, it is assumed the antibody-antigen exists in the mixture. The complement system described by Erhlich formed the basis for the complement fixation test which has been used as an indirect method to discover "novel viruses" or to determine someone positive for a known "virus." Here is a brief description of what this test entails:

"The complement fixation test consists of two components.

The first component is an indicator system that USES COMBINATION OF SHEEP RED BLOOD CELLS, COMPLEMENT-FIXING ANTIBODY SUCH AS IMMUNOGLOBULIN G PRODUCED AGAINST THE SHEEP RED BLOOD CELLS AND AN EXOGENOUS SOURCE OF COMPLEMENT USUALLY GUINEA PIGS SERUM. When these elements are MIXED IN OPTIMUM CONDITIONS, the anti-sheep antibody binds on the surface of red blood cells. Complement subsequently binds to this antigen -antibody complex formed and will cause the red blood cells to lyse.

The second component is Test System (A KNOWN ANTIGEN AND PATIENT SERUM ADDED TO A SUSPENSION OF SHEEP RED BLOOD CELLS IN ADDITION TO COMPLEMENT). These two components of the complement fixation method are tested in sequence. Patient serum is first added to the known antigen, and complement is added to the solution. If the serum contains antibodies to the antigen, the resulting antigen-antibody complexes will bind all of the complement. SHEEP RED BLOOD CELLS AND THE ANTI-SHEEP ANTIBODY ARE THEN ADDED. If complement has not been bound by an antigen-antibody complex formed from the patient serum and known antigens, it is available to bind to the indicator system of sheep cells and anti-sheep antibodies. Lysis of the indicator sheep red blood cells signifies both a lack of antibody in patient serum and a negative complement fixation test. If the patient’s serum does contain a complement-fixing antibody, A POSITIVE RESULT WILL BE INDICATED BY THE LACK OF RED BLOOD CELL LYSIS."

https://bio.libretexts.org/.../12.2G%3A_Complement_Fixation

There are some noted drawbacks for the test beyond the combining of human blood with that from Sheep/Guinea pigs:

"DISADVANTAGES OF COMPLEMENT FIXATION TEST

NOT SENSITIVE – cannot be used for immunity screening.

Time-consuming.

OFTEN NON-SPECIFIC e.g. CROSS-REACTIVITY between Herpes Simplex Virus and Voricella Zoster Virus."

https://microbenotes.com/complement-fixation-test-steps-advantages-and-disadvantages

So the test, based on the theoretical theory proposed by Ehrlich, is not sensitive nor is it specific and often leads to cross-reactivity with other "viruses." This obviously poses a problem as in order to know which antibody reacts and is specific for a "virus" in order to detect it, one would have to have first purified/isolated an actual "virus" directly from a human sample. Without this, sensitivity and specificity can not be concluded nor can specific antibodies be known.

This is the problem with basing tests on theoretical concepts and experimental reactions. To date, the antibody-antigen relationship is still nothing but an unproven theory. This was brilliantly summarized in the paper EHRLICH'S "BEAUTIFUL PICTURES:"

"The term antibody today refers to discrete biochemical entities present in the blood.

THE BELIEF THAT ANTIBODIES ARE SUCH ENTITIES is held not only by scientists and physicians, but also by the lay public, who learns about "antibodies," if not in school, then at least in the course of routine medical practices such as vaccination and, increasingly, through the so-called popularization of science. In addition, some people will readily associate the term antibody with the characteristic Y-shaped structure found not only in textbooks and specialized scientific articles but also, more recently, in advertisements and even as the logos of pharmaceutical and biotechnology companies. In order to understand the beginnings of immunological imagery at the turn of the century, WE HAVE TO DISCARD OUR PRESENT-DAY NOTIONS ABOUT THE REALITY AND STRUCTURE OF ANTIBODIES."

"THE DEBATE OVER THE EXISTENCE, NATURE, AND-PROPERTIES OF "ANTIBODIES" LASTED FOR DECADES. Very few people shared the extreme position of Felix Le Dantec, who DENIED THE PHYSICAL EXISTENCE OF WHAT HE CALLED "PHENOMENINES"-that is, MYTHICAL SUBSTANCES that, like Moliere's "vertu dormitive," WERE BEING USED TAUTOLOGICALLY TO ACCOUNT FOR EMPIRICALLY OBSERVED PHENOMENA:

'While making fun of the physicians of his time, our great Moliere had foreseen Ehrlich's

system. EHRLICH HAS ADDED NOTHING TO THE EXPLANATION OF THE IMAGINARY INVALID. Rather than saying that chloral causes sleep because it contains a soporific power ("vertu dormitive"), we would today say, according to the German scholar, that chloral contains a soporine ("dormitine"); well, it is the same thing! . . . Here then is a therapeutic serum which produces, when inoculated into the rabbit, a given phenomenon. How will you

explain this particular activity? It is very simple: put on your glasses and your doctor's

cap and gravely say: "THE SERUM PRODUCES THIS PHENOMENON BECAUSE IT CONTAINS A PHENOMENINE WHICH HAS THAT POWER." Nobody will laugh. Point out timidly that the same serum has a different action on the snake; that it produces a second phenomenon; that will be because it contains a second phenomenine. If the same substance produces a thousand different phenomena in a thousand different species, then it contains a thousand

different phenomenines; and there you go! Why had we not thought of this earlier?

HENCEFORTH WE HAVE THE EXPLANATION FOR EVERYTHING!'

"Moreover, other researchers, while not sharing Le Dantec's extreme position, did share his hostility to "terminology-based" explanations. Henry Dean, for instance, noted that "agglu-

tinins, precipitins, amboceptors ARE MERE WORDS, and A PASSIVE BELIEF IN THE EXISTENCE OF SUCH BODIES TENDS TO IMPEDE RATHER THAN ADVANCE OUR UNDERSTANDING OF WHAT IS ACTUALLY TAKING PLACE"; he added that "IGNORANCE, HOWEVER APTLY VEILED IN AN ATTRACTIVE TERMINOLOGY, REMAINS IGNORANCE."

"So, DESPITE VARIOUS PROFESSIONS OF FAITH in the material nature of "antibodies," their ontological status remained uncertain, a situation ascribed by some scientists to THE FAILURE TO PURIFY CHEMICALLY THE ELUSIVE ENTITIES AND THUS TO ASCERTAIN WHETHER THEY WERE INDEED MATERIAL SUBSTANCES."

"In a 1902 letter nominating Ehrlich for the Nobel Prize, Bernhard Naunyn noted that the German researcher could be credited with the introduction of a stereochemical approach into biology and could thus be compared with the great scientists (August Kekule, Adolf von Baeyer) who had done the same in their own disciplines; BUT HE ADDED THAT EHRLICH'S CONTRIBUTION SHOULD STILL BE REGARDED AS TENTATIVE OR PREMATURE, SINCE THE ISOLATION AND PURIFICATION OF THE RELEVANT SUBSTANCES (namely, "ANTIBODIES") WOULD LONG REMAIN A CHIMERA. Those same substances were treated, in a 1910 German textbook, as didactic devices:

'In order to learn the nature of these antibodies attempts have been made to isolate them

chemically. THUS FAR ALL SUCH TRIALS HAVE BEEN UNSUCCESSFUL. IT IS EVEN UNCERTAIN WHETHER THESE SO-CALLED ANTIBODIES ARE DEFINITE CHEMICAL ENTITIES. Only the effects of the serum as a whole are known, AND THE INGREDIENTS IN IT TO WHICH THESE ACTIVITIES ARE ATTRIBUTED ARE THOUGHT OF AS ANTIBODIES. For didactic purposes antibodies, as antitoxins, agglutinins, etc., will be spoken of in this book when the antitoxin or agglutinating properties, exclusively, are meant. 15'

Julius Citron's 1910 statement was echoed nineteen years later by Wells, according to whom:

We attribute this altered reactivity [of sera] to the presence of "antibodies," DESPITE THE

FACT THAT WE HAVE ABSOLUTELY NO KNOWLEDGE OF WHAT THESE ANTIBODIES MAY BE, OR EVEN THAT THEY EXIST AS MATERIAL OBJECTS. Like the enzymes, WE RECOGNIZE THEM BY WHAT THEY DO WITHOUT DISCOVERING JUST WHAT THEY ARE. We do not know whether they are specific molecular aggregates OR MERELY PHYSICAL FORCES DEPENDENT ON ALTERED SURFACE ENERGY OF THE SAME SUBSTANCES PRESENT IN THE BLOOD BEFORE THE PROCESS OF IMMUNIZATION WAS BEGUN."

https://www.jstor.org/stable/235104

Ehrlich provided a theory that weaved together various disparate elements like the best fictions writers do. He defined the concepts of antibody, antigen, the complement system, and provided a framework for how immunity works. This led to indirect non-specific and non-sensitive complement fixation tests used as evidence for the existence of "viruses" and/or as proof that the vaccines are effective against them. The problem is that Ehrlich's theories were just mere words with nothing physical backing them. He created concepts and ideas for that which he could not physically see. They belong to the realm of fantasy and fairy tales.

As Henry Dean stated: "ignorance, however aptly veiled in an attractive terminology, remains ignorance."

Part 1:

https://docs.google.com/document/d/e/2PACX-1vSWQ7pM49P09Qvdn1pj20KLoOCyX4Oyy5ShBJUWHGz5CQAqYrcLT4iGostVXFwA76qmKWp3ig2kh0uj/pub

______

ASTRID FAGRAEUS 1948 PLASMA CELLS PRODUCE ANTIBODIES CORRELATION:

A "breakthrough" in Immunology came in 1947/48 when Astrid Fagraeus published her doctoral dissertation "Antibody Production in relation to the Development of Plasma Cells." She is given the credit for showing that the plasma cells produce antibodies. Before her paper, their function was unknown. But did Astrd Fagraeus really prove the correlation between plasma cells and antibodies definitively?

According to the book "Advances in Immunology" by Frederick W. Alt (Image # 3 below):

(My paraphrasing)

In 1938, Fred Kolouch Jr. directly correlated the formation of plasma cells with the injection of an antigen. From this correlation, he postulated that plasma cells produce antibodies even though he had no direct experimental evidence backing up his hypothesis. It wasn't until 1948, with the experiments of Astrid Fagraeus, where a breakthrough in Immunology occurred. She performed in vitro studies with plasma cell-enriched spleen biopsies from horse serum-injected rabbits and found a correlation between the numbers of plasma cells in the tissues and the amount of secreted antibodies in the culture medium. However, even with her findings, she was unable to provide clear evidence that the plasma cells were in fact the cells that secrete antibodies.

However, the lack of clear evidence did not stop the scientific community from recognizing Fagraeus and her findings as proof that plasma cells produce the still unseen antibodies:

"Fagraeus' doctoral dissertation, 'Antibody Production in relation to the Development of Plasma Cells', attracted international attention and was CONSIDERED A MILESTONE IN MODERN IMMUNOLOGY. In this work, SHE WAS THE FIRST TO SHOW THAT PLASMA CELLS PRODUCE ANTIBODIES (IgG)."

https://en.m.wikipedia.org/wiki/Astrid_Fagraeus

"It took another 60 years until ASTRID FAGREUS DISCOVERED THE CELLS SECRETING THOSE ANTIBODIES, THE “PLASMA CELLS”, describing them as “terminal stages of B cell differentiation”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3337994/

"In 1943, Mogens Bjørneboe and Harald Gormsen were the first to experimentally show that repeated immunization of rabbits with polyvalent vaccines leads to massive proliferation of plasma cells in most organs and that THIS PROLIFERATION CORRELATES WITH ANTIBODY CONCENTRATION.

That finding was supported a few years later by Astrid Fagraeus, who reported that plasma cells produce antibodies in vitro. Tissue cultures of spleens from rabbits immunized with live bacteria showed abundant formation of plasma cells. FAGRAEUS CONCLUDED THAT PLASMA CELLS APPEAR IN CONNECTION WITH STRONG ANTIGEN STIMULATION."

https://www.nature.com/milestones/mileantibodies/full/mileantibodies03.html

The actual paper was difficult to find but I did manage to find and take an image of a Nature article in which she details her work (Image # 2 below) as well a source which provided a brief summary:

THE PLASMA CELLULAR REACTION AND ITS RELATION TO THE FORMATION OF ANTIBODIES IN VITRO

The Journal of Immunology 58 (1), 1-13, 1948

"1.During secondary response, elicited in sensitized rabbits by means of intravenous injections of antigen, A GREAT INCREASE IN THE NUMBER OF PLASMA CELLS (pl.c.) IN THE SPLEEN WAS RECORDED SIMULTANEOUSLY WITH THE INCREASE OF CIRCULATING ANTIBODIES.

2.Pl.c. were confined almost exclusively to the red pulp, especially after the injection of living S. typhi. They originated apparently from reticulum cells, passing through a chain of development: transitional cell → immature pl.c. → mature pl.c.

3.Pieces of spleen were excised at different times during the period of antibody formation in rabbits, DIFFERENTIAL CELL COUNTS were made and the capacity of excised splenic tissue to form antibodies in vitro was investigated. The following observations were made:

a.THE AMOUNT OF ANTIBODY LIBERATED IN PLAIN TISSUE EXTRACTS, under conditions preventing growth or metabolism of cells, WAS VERY LOW, WHEREAS SIGNIFICANT YIELDS WERE OBTAINED IN TISSUE CULTURES.

b.The capacity of the red pulp abundant in pl.c. to produce antibodies in vitro was considerably superior to that of lymph follicles, rich in lymphocytes but devoid of pl.c.

c.Antibody production was comparatively poor in tissue containing only transitional cells, reached a maximum when numerous immature pl.c. were present and receded when predominantly mature pl.c. were found.

4.After intravenous injections the antigen accumulated in those places where pl.c. subsequently developed.

The conclusion is drawn that ANTIBODIES UNDER THE CONDITIONS OF THE EXPERIMENTS, are formed by cells of the R.E.S., passing through a chain of development, the final link of which is the mature pl.c."

https://www.jimmunol.org/content/58/1/1.short

doi: 10.1038/159499a0.

It appears that the scientific community is very much of the mindset that correlation equals causation. They seem to forget this very basic premise:

"Can correlation ever equal causation?

Correlation tests for a relationship between two variables. HOWEVER, SEEING TWO VARIABLES MOVING TOGETHER DOES NOT NECESSARILY MEAN WE KNOW WHETHER ONE VARIABLE CAUSES THE OTHER TO OCCUR. This is why we commonly say “CORRELATION DOES NOT IMPLY CAUSATION.”

https://www.jmp.com/en_us/statistics-knowledge-portal/what-is-correlation/correlation-vs-causation.html

_______

MICHAEL HEIDELBERGER 1929 ANTIBODY EQUATION:

Michael Heidelberger is considered the "Founder of Immunochemistry." He was the first to apply mathematics to the reaction of antibodies and their antigens. He is also known for "proving" that antibodies are proteins by showing that the antigens of pneumococcus bacteria are polysaccharides (or carbohydrates). Here is a brief overview of his work:

"Heidelberger and Avery's discovery came at a time when ANTIBODIES WERE REGARDED—BY THOSE WHO BELIEVED THEY EXISTED AT ALL—AS MYSTERIOUS SUBSTANCES THAT FLOATED AROUND IN SERUM. “It appeared to me that there was a crying need to determine the true nature of antibodies,” wrote Heidelberger in 1979, “and that until this was done there could be no end to the polemics and uncertainties that were plaguing immunology” (2). Heidelberger later purified the antibodies from his precipitin reactions and showed that they themselves were proteins. As a result, says friend and colleague Victor Nussenzweig (New York University), “THERE WERE NO MORE MYSTICAL IDEAS ABOUT WHAT ANTIBODIES WERE.”

Heidelberger and his postdoctoral fellow Forrest Kendall later quantitated the precipitin reaction (5), BRINGING MUCH-NEEDED MATHEMATICS TO THE STUDY OF ANTIBODY–ANTIGEN INTERACTIONS AND LIFTING ANTIBODIES EVEN FURTHER OUT OF THE REALM OF THE MYSTERIOUS (see the next “From the Archive”).

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2212983

Two of Heidelberger's papers are most often cited as the proof that antibodies are proteins. The first is a paper he did with Oswald Avery in 1923. Some highlights below:

THE SOLUBLE SPECIFIC SUBSTANCE OF

PNEUMOCOCCUS.

"In 1917 Dochez and Avery (1) showed that WHENEVER PNEUMOCOCCI ARE GROWN IN FLUID MEDIA, THERE IS PRESENT IN THE CULTURAL FLUID A SUBSTANCE which precipitates specifically in antipneumococcus serum of the homologous type. This soluble substance is demonstrable in culture filtrates during the initial growth phase of the organisms; that is, during the period of their maximum rate of multiplication

when little or no cell death or disintegration is occurring. THE FORMATION OF THIS SOLUBLE SPECIFIC MATERIAL BY PNEUMOCOCCI ON GROWTH IN VITRO SUGGESTED THE PROBABILITY OF AN ANALOGOUS SUBSTANCE BEING FORMED ON GROWTH OF THE ORGANISM IN THE ANIMAL BODY. Examination of the blood and urine of experimentally infected animals gave proof of the presence of this substance in considerable quantities in the body fluids following intraperitoneal infection with pneumococcus. In other words, this soluble material elaborated at the focus of the disease readily diffuses throughout the body, is taken up in the blood, passes the kidney, and appears in the urine unchanged in specificity. Similarly, a study of the serum of patients suffering from lobar pneumonia has revealed a substance of like nature in the circulating blood during the course of the disease in man. Furthermore, EXAMINATION OF THE URINE OF PATIENTS having pneumonia due to pneumococci of Types I, II, and III HAS SHOWN THE PRESENCE OF THIS SUBSTANCE IN SOME STAGE OF THE DISEASE IN APPROXIMATELY TWO-THIRDS OF THE CASES. Recently from filtered alkaline extracts of pulverized bacteria of several varieties, including pneumococci, Zinsser and Parker (2) have prepared substances which appear free from coagulable protein. These substances, called "residue antigens," are specifically predpitable by homologous antisera. THESE OBSERVERS CONSIDER THESE ACID- AND HEAT-RESISTANT ANTIGENIC MATERIALS ANALOGOUS TO THE SOLUBLE SPECIFIC SUBSTANCE OF PNEUMOCOCCUS described by Dochez and Avery (1). In spite of the fact that these "residue antigens" are predpitable by homologous sera produced by immunization with the whole bacteria, Zinsser and Parker HAVE SO FAR FAILED TO PRODUCE ANTIBODIES IN ANIMALS BY INJECTING THE RESIDUES."

"EXPERIMENTAL

The organism used in the present work was Pneumococcus Type II. The most abundant source of the soluble specific substance

appeared to be an 8 day autolyzed broth culture; hence this material was used as the principal source of supply. For comparison dis-

solved pneumococci and lots of urine containing the specific substance

were also worked up, with essentially the same results, as will be seen from Table I.

The process for the isolation of the soluble specific substance consisted in concentration of the broth, precipitation with alcohol,

repeated re-solution and reprecipitation, followed by a careful series of fractional precipitations with alcohol or acetone after acidification of the solution with acetic acid, and, finally, repeated fractional precipitation with ammonium sulfate and dialysis of the aqueous solution of the active fractions."

"THE RESIDUAL MATERIAL (Preparation 17, in Table I), FOR WHICH NO CLAIM OF PURITY IS MADE, AS EFFORTS AT ITS FURTHER PURIFICATION ARE STILL UNDER WAY, contained, on the ash-free basis, 1.2 per cent of nitrogen. IT WAS ESSENTIALLY A POLYSACCHARIDE, as shown by the formation of 79 per cent of reducing sugars on hydrolysis, and by the isolation and identification of glucosazone from the products of hydrolysis."

"ATTEMPTS TO STIMULATE ANTIBODY PRODUCTION BY THE IMMUNIZATION OF ANIMALS WITH THE PURIFIED SUBSTANCE YIELDED NEGATIVE RESULTS."

"DISCUSSION.

While it has long been known that the capsular material of many microorganisms consists, at least in part, of carbohydrates (4), any connection between this carbohydrate material and the specificity relationships of bacteria appears to have remained unsuspected. WHILE IT CANNOT BE SAID THAT THE PRESENT WORK ESTABLISHES THIS RELATIONSHIP, it certainly points in this direction. Evidence in favor of the probable carbohydrate nature of the soluble specific substance is the increase in specific activity with reduction of the nitrogen content, the increase in optical rotation with increase in specific activity, the parallelism between the Molisch reaction and specific activity, the high yield of reducing sugars on hydrolysis, and the actual isolation of glucosazone from a small quantity of the material. The small amounts of substance available up to the present have hindered the solution of the problem, and it is hoped that efforts at further purification of the soluble specific substance, now in progress with larger amounts of material, will definitely settle the

question.

SUMMARY

1. A method is given for the concentration and purification of the soluble specific substance of the pneumococcus.

2. The material obtained by this method is shown to consist mainly of a carbohydrate which appears to be a polysaccharide built up of glucose molecules.

3. WHETHER THE SOLUBLE SPECIFIC SUBSTANCE IS ACTUALLY THE POLYSACCHARIDE, OR OCCURS MERELY ASSOCIATED WITH IT, IS STILL UNDECIDED, although the evidence points in the direction of the former possibility."

https://doi.org/10.1084/jem.38.1.73

In Summary:

-whenever pneumococci is grown in culture fluid media, a soluble specific substance is found

-they assume that this growth is culture suggests that this also same substance occurs inside the animal body

-examination of the urine of patients with pneumococci showed the substance in only 2/3rds of the samples

-some other researchers found similar substances with other bacteria and believe they are the same as that of the pneumococci

-injecting these substances in animals has FAILED to produce antibodies

-they make no claim as to the purity of the material in their experiments and state further purification is being attempted

-they believe it is ESSENTIALLY a polysaccharide

-they were also UNABLE to stimulate antibodies by injecting their substance in animals

-they state that their work CAN NOT ESTABLISH a relationship between bacteria and carbohydrates

-whether the soluble specific substance is a polysaccharide or occurs merely associated with it was left undetermined

This paper doesn't seem to be the slam-dunk proof that bacteria antigens are carbohydrates so it seems rather odd to assume antibodies are proteins based off this work, but assume they did:

"Since the pneumococcal capsular antigen was a polysaccharide, AND ANTIBODIES WERE THOUGHT TO BE PROTEINS, Heidelberger realized that by MEASURING THE AMOUNT OF PROTEIN IN SPECIFIC PRECIPITATES MADE WITH THE CAPSULAR ANTIGEN HE COULD DETERMINE THEIR ANTIBODY CONTENT. Together with Forrest Kendall, who had joined the Heidelberger lab, THE PROTEIN CONTENT OF IMMUNE PRECIPITATES WAS DETERMINED BY MEASURING TOTAL NITROGEN, using the Kjeldahl procedure that came to be the hallmark of laboratories carrying out Heidelberger-type quantitative immunochemistry."

https://www.google.com/url?sa=t&source=web&rct=j...

Since they assumed the pneumococcal bacteria was a polysaccharide, that meant any nitrogen left over was the antibody content. This second paper shows how Heidelberger came to this conclusion using the precipitin test (images below) and mathematics as proof that antibodies exist. I edited out the long mathematical sections with his equations so if you are interested in Heidelberger showing his work, I recommend reading the full paper. Highlights below:

A QUANTITATIVE STUDY OF THE PRECIPITIN REACTION BETWEEN TYPE III PNEUMOCOCCUS POLYSACCHARIDE AND PURIFIED HOMOLOGOUS ANTIBODY

"Of all the reactions of immunity the precipitin test is perhaps the most dramatic and striking. While other immune reactions are more delicate, the precipitin test is among the most specific and least subject to errors and technical difficulties. ATTEMPTS AT ITS QUANTITATIVE INTERPRETATION AND EXPLANATION (1, 2) HAVE BEEN HAMPERED EITHER BY THE DIFFICULTY OF FINDING SUITABLE ANALYTICAL METHODS OR BY THE FAILURE TO SEPARATE THE REACTING SUBSTANCES FROM CLOSELY RELATED, NON-SPECIFIC MATERIALS WITH WHICH THEY ARE NORMALLY ASSOCIATED.

With the aid of recent work IT HAS BEEN FOUND POSSIBLE TO AVOID THESE DIFFICULTIES TO SOME EXTENT. The isolation of bacterial polysaccharides which precipitate antisera specifically (3) and possess the properties of haptens (4) has not only afforded one of the components of a precipitin reaction IN A STATE OF COMPARATIVE PURITY, but has greatly simplified the analytical problem. Since many of these polysaccharides contain no nitrogen, AND ANTIBODIES PRESUMABLY ARE NITROGENOUS, the latter may be determined in the presence of any amount of the specific carbohydrate. Moreover, Felton's method for the separation of pneumococcus antibodies from horse serum (5) not only permits the isolation of a high proportion of the precipitin, FREED FROM AT LEAST 90 PERCENT OF THE SERUM PROTEINS AND MUCH OF THE SERUM LIQUID, but is also applicable on a sufficiently large scale to furnish the amounts of antibody solution needed to make quantitative work possible. IT IS REALIZED THAT ANTIBODY SOLUTIONS OF THIS TYPE DO NOT CONTAIN PURE ANTIBODIES--INDEED, ONLY 40 TO 50 PERCENT OF THE NITROGEN IS SPECIFICALLY PRECIPITABLE-- BUT SINCE SO SMALL A PROPORTION OF THE ORIGINAL SERUM PROTEIN REMAINS WITH THE ANTIBODY a far-reaching purification actually has been effected. It should thus be possible with the aid of antibodies purified by Felton's method to OBTAIN DATA OF A PRELIMINARY CHARACTER which should point toward the mechanism of the reaction. The present paper is concerned with such data obtained in a quantitative study of the precipitin reaction between the soluble specific substance of Type III pneumococcus and Type III pneumococcus antibody solution.

EXPERIMENTAL

1. Materials and Methods.--a. Solutions of Soluble Specific Substance, Type Ill

Pneumococcus.--The soluble specific substance of Type III pneumococcus (6)* used was kinky supplied by Drs. O. T. Avery and W. F. Goebel of The Rockefeller Institute for Medical Research. It was ash-free, contained 0.04 per cent of nitrogen, and showed [aid = -32 °. A weighed amount of anhydrous substance was suspended in 0.9 per cent saline, dissolved with the aid of 0.1 normal sodium hydroxide, and the solution was diluted with saline, adjusted to pH 7.6 and made up to volume with saline to yield a 1 per cent solution. This was sterilized in the autoclave and used as a stock solution for making up other dilutions. These were prepared with sterile saline under aseptic precautions, and were kept in the ice-box.

b. Type III Pneumococcus Antibody Solution.--The antibody solutions used were prepared essentially according to Felton's procedure (loc. cir.) from Type III anti pneumococcal horse serum containing no preservative and supplied by the New York State Department of Health through the courtesy of Dr. A. B. Wadsworth and Dr. Mary B. Kirkbride. 100 to 200 cc. of serum were stirred slowly into 20 volumes of ice-cold water containing 9.5 cc. of molar potassium dihydrogen phosphate and 0.5 cc. of molar dipotassium hydrogen phosPhate per liter. The final pH varied from 5.6 to 6.3. Mter standing overnight in the cold the supernatant was decanted and the precipitate was centrifuged off in the coldt and dissolved in a volume of chilled 0.9 percent saline equal to that of the serum taken. 0.1 normal hydrochloric acid was then added until a precipitate no longer formed on dilution of a test portion with two volumes of water, after which 0.1 normal sodium hydroxide solution was added until a slight precipitate again formed on dilution. In general, 5 cc. of acid and 1.5 cc. of alkali per 100 cc. of serum were satisfactory, although as Felton emphasizes, DIFFERENT LOTS VARY AND NO ABSOLUTELY DEFINITE PROCEDURE CAN BE GIVEN. In the present work the process of purification was followed either by testing the agglutinating power of the fractions against a heat-killed Type III pneumococcal vaccine, or by the precipitin reaction, or by both methods. After addition of the alkali the opalescent solution was diluted with 2 volumes of water and centrifuged in the cold. The almost inactive precipitate was discarded and the supernatant poured into 6.7 volumes of the chilled buffer solution previously used, (equivalent to 20 times the volume of saline employed), also adding enough 0.1 normal sodium hydroxide to neutralize the remaining acid. The resulting precipitate was collected and dissolved in a volume of 0.9 per cent saline equal to that of the serum taken, and the pH was adjusted to 7.6. The solution was sterilized by passage through a Berkefeld N grade filter which previously had been washed with saline containing a drop of normal sodium hydroxide, followed by saline alone.

ANTIBODY SOLUTIONS PREPARED IN THIS WAY WERE FOUND TO BE RATHER UNSTABLE UNDER THE USUAL CONDITIONS OF THE PRECIPITIN TEST, AND IT THEREFORE WAS NECESSARY TO SUBJECT THEM TO A PRELIMINARY "AGEING" TREATMENT in order that control solutions might be relied upon to remain clear. This consisted in immersing the solution in a water bath at 37 ° for 2 hours, letting it stand in the ice-box overnight, centrifuging off the precipitate which usually formed, readjusting the pH if necessary, and filtering through a Berkefeld candle prepared as above. THIS TREATMENT WAS REPEATED AS MANY TIMES AS NECESSARY, but the solutions usually remained clear after the second incubation at 37 ° . MUCH TIME WAS LOST AND VERY INCONSISTENT RESULTS WERE OBTAINED UNTIL "AGEING" WAS RESORTED TO.*

THE RELATIVE ANTIBODY CONTENT OF THE RESULTING SOLUTIONS WAS ESTIMATED by determining the agglutination titer against a single heat-killed Type III pneumococcus suspension."

"DISCUSSION

For purposes of discussion IT WILL BE ASSUMED WITH FELTON (lot. cir.) THAT ANTIBODY IS MODIFIED PROTEIN, AND THAT, IN ORDER TO PROVIDE A UNIFORM METHOD OF MEASUREMENT, IT MAY BE EXPRESSED AS NITROGEN PRECIPITATE BY SPECIFIC POLYSACCHARIDE, MULTIPLIED BY 6.25. Since only relative values are under consideration, the actual magnitude of the factor used is of little significance so long as it be used throughout.

Moreover, Table I shows a correspondence between this measure of antibody content and the agglutination titer, so that its use as a relative measure is independent of the nature of Type III pneumococcal antibodies."

"WHETHER OR NOT THESE CONCEPTIONS ARE OF GENERAL APPLICATION REMAINS TO BE TESTED, and work along these lines is under way."

doi: 10.1084/jem.50.6.809.

In Summary:

-Heidelberger acknowledges that the precipitin test used during this experiment has 2 drawbacks:

a) quantitative interpretation/explanation is difficult due to lack of a suitable analytical method

b) failure to separate out the reacting substances from non-specific material which these substances are closely related to and associated with

-he states it is possible to avoid these failures TO SOME EXTENT

-he PRESUMES antibodies are nitrogenous

-he states at least 90% of the precipitin can be freed from serum proteins and liquid

-Heidelberger admits that these are NOT PURE ANTIBODIES and that only 40-50% of nitrogen is precipitable while small amounts of serum remain

-antibodies were found to be unstable during testing so they were put through preliminary "ageing" processes as many times as needed until they got the result they wanted

-much time was lost and results were inconsistent until the 'ageing" process was implemented

-the antibody content was then ESTIMATED

--since Heidelberger thought pneumococci antigens were carbohydrate, he ASSUMED that meant the antibodies were protein and thus any nitrogen left over after the precipitin test contained the antibodies

-whether or not his conceptions were of general application remained to be tested

It seems rather obvious that many assumptions were made about a substance (antibodies) for which they could not see. The conclusions drawn were born out of chemistry experiments which have no bearing on reality and mathematical equations attempting to quantify the unquantifiable. Whether or not these indirect experiments and assumptions provide proof that antibodies exist and are proteins, I leave up to the reader. However, keep in mind that no antibodies had ever been seen nor proven to exist up to that time (and to date). This work is based off of theoretical explanations of immunity for which nothing could be observed.

One last interesting fact. Remember the pneumococci bacteria they based their "antigen" on and for which they could not produce antibodies with when testing on animals? It is regularly found in healthy people.

"Infection is acquired mainly through pneumococci contained in respiratory droplets. THERE ARE MANY HEALTHY, ASYMPTOMATIC CARRIERS OF THE BACTERIA but no animal reservoir or insect vector."

https://www.who.int/ith/diseases/pneumococcal/en/

If an antigen is a toxin or foreign substance which produces an immune response creating antibodies, the pneumococci bacteria doesn't seem to meet that definition at all.

_________

FRANK MACFARLANE BURNET 1957 CLONAL SELECTION THEORY:

"A SCIENTIFIC THEORY is not the end result of the scientific method; THEORIES CAN BE PROVEN OR REJECTED, JUST LIKE HYPOTHESES. Theories can be improved OR MODIFIED as more information is gathered so that the accuracy of the prediction becomes greater over time."

https://www.livescience.com/21491-what-is-a-scientific-theory-definition-of-theory.html

At the time that Frank M. Burnet proposed the Clonal Selection Theory of antibody formation in 1957, there were a few different theories floating about. They were the:

1. Direct Template Theory

2. Indirect Template Theory

3. Side Chain Theory

4. Natural Selection Theory

Each of these theories proposed different ways antibodies formed based on the scientific evidence of the time. Naturally, Burnet saw the burning need for a new theory to be thrown into the mix which is why he utilized Jerne's Natural Selection Theory to rework and create another one of his own. Thus the Clonal Selection Theory was born and it has gone on to become the most widely accepted explanation for the formation of antibodies, which were still unseen particles assumed to be within the blood:

"Clonal selection theory is a scientific theory in immunology that explains the functions of cells of the immune system (lymphocytes) in response to specific antigens invading the body. The concept was introduced by Australian doctor Frank Macfarlane Burnet in 1957, in an attempt to explain the great diversity of antibodies formed during initiation of the immune response.[1][2] THE THEORY HAS BECOME THE WIDELY ACCEPTED MODEL FOR HOW THE HUMAN IMMUNE SYSTEM RESPONDS TO INFECTION and how certain types of B and T lymphocytes are selected for destruction of specific antigens.[3]"

https://en.m.wikipedia.org/wiki/Clonal_selection

"THE CLONAL SELECTION HYPOTHESIS HAS BECOME A WIDELY ACCEPTED MODEL for how the immune system responds to infection and how certain types of B and T lymphocytes are selected for destruction of specific antigens invading the body."

https://bio.libretexts.org/Bookshelves/Microbiology/Book%3A_Microbiology_(Boundless)/11%3A_Immunology/11.07%3A_Antibodies/11.7C%3A_Clonal_Selection_of_Antibody-Producing_Cells

"In 1957, Burnet proposed that central tolerance to self-antigens occurred by clonal deletion of self-reactive lymphocytes. However, as we will discuss, THIS MODEL HAS BEEN ‘UPDATED’ ON SEVERAL OCCASIONS."

https://www.cell.com/trends/immunology/fulltext/S1471-4906(04)00311-4

Notice the MODEL word thrown about in each of those descriptions? What could that possibly be referring to?

"SCIENTIFIC MODELING, the generation of a physical, CONCEPTUAL, or mathematical REPRESENTATION of a real PHENOMENON THAT IS DIFFICULT TO OBSERVE DIRECTLY."

https://www.britannica.com/science/scientific-modeling

This Clonal Theory, which can be rejected just like any Hypothesis, is the current Model, which is a conceptual representation of a phenomenon that is DIFFICULT TO OBSERVE DIRECTLY. The previous theories/models of this unseen phenomenon were unceremoniously thrown out in favor of Burnet's Clonal Theory. However, even Clonal Theory is being challenged today, but I digress. Burnet's entire theory is presented below:

A MODIFICATION OF JERNE'S THEORY OF ANTIBODY PRODUCTION USING THE CONCEPT OF CLONAL SELECTION

"THERE ARE THREE CURRENT THEORETICAL INTERPRETATIONS OF ANTIBODY PRODUCTION which, following Talmage (1957), may be referred to as the DIRECT TEMPLATE THEORY in which the antigen serves as a template against which the specific pattern of the antibody is synthesized, the INDIRECT TEMPLATE THEORY which postulates a secondary template incorporated into the genetic-synthetic processes of the antibody producing cells (Burnet,1956), and the NATURAL SELECTION THEORY in which the antigen acts essentially by selection for excess production of natural antibody molecules of corresponding type (Jerne, 1955).

THE TWO LATTER THEORIES WERE DEVISED PRIMARILY TO ACCOUNT FOR TWO SETS OF PHENOMENA FOR WHICH THE DIRECT TEMPLATE THEORY SEEMS QUITE IRRELEVANT. The first is the absence of immunological response to "self" constituents and the related phenomena of immunological

tolerance; the second is the evidence that

antibody production can continue in the absence of antigen. Some means for the

recognition and differentiation of potentially antigenic components of the body from foreign organic material must be provided in any acceptable formulation.

In Burnet and Fenner's (1949) account, a positive recognition of "self" material was ascribed to the presence of "self markers" in all potentially antigenic macromolecules, and corresponding recognition units in the scavenger cells of the body. AT THE TIME IT WAS REGARDED AS INCONCEIVABLE THAT A MECHANISM COULD EXIST WHICH WOULD RECOGNIZE IN POSITIVE FASHION ALL FOREIGN MATERIAL AND NO ATTEMPT WAS MADE TO DEVISE ONE, despite the fact that we have always recognised the clumsy character of the self-marker, self-recognition scheme.

IT IS THE GREAT VIRTUE OF JERNE'S HYPOTHESIS THAT IT PROVIDES AN APPROACH TO THIS ALTERNATIVE METHOD OF RECOGNIZING SELF FROM NONSELF. There is no doubt about the presence in all mammalian or avian sera of a wide range of reactive globulins which can legitimately be called "natural antibodies." JERNE ASSUMED THAT AMONGST THESE GLOBULIN MOLECULES WERE ALL THE POSSIBLE PATTERNS NEEDED FOR SPECIFIC IMMUNOLOGICAL TYPE REACTIONS WITH ANY ANTIGEN, except for those patterns corresponding to body antigens which would be eliminated by in vivo absorption. When a foreign antigen enters the blood it unites, according to Jerne's scheme, with one of the corresponding natural antibody molecules. The complex is taken up by a phagocytic cell in which the antigen plays no further part, but the antibody globulin provokes the production by the cell of a fresh crop of similar molecules which are liberated as antibodies. IF THIS BASIS IS ACCEPTED, MOST IMMUNOLOGICAL PHENOMENA CAN BE WELL DESCRIBED IN TERMS OF THE THEORY. Its major objection is the absence of any precedent for, and the intrinsic unlikelihood of, the suggestion that a molecule of partially denatured antibody could stimulate a cell, into which it had been taken, to produce a series of replicas of the molecule.

TALMAGE (1957) HAS SUGGESTED THAT JERNE'S VIEW IS BASICALLY AN EXTENSION OF EHRLICH' SIDE CHAIN THEORY of antitoxin production and that it would be more satisfactory if the replicating elements essential to any such theory were cellular in character ab initio rather than extracellular protein which can replicate only when taken into an appropriate cell. TALMAGE DOES NOT ELABORATE THIS POINT OF VIEW BUT CLEARLY ACCEPTS IT AS THE BEST BASIS FOR THE FUTURE DEVELOPMENT OF ANTIBODY THEORY. He stresses the multiplicity of the globulin types that can be present in the blood and is profoundly sceptical of any approach which attempts to "unitarian" an interpretation of antibody. In his view properdin has as much right to be called an antibody as any other globulin.

Before receiving Talmage's review we had adopted virtually the same approach but had developed it from what might be called a "clonal" point of view. THIS IS SIMPLY A RECOGNITION THAT THE EXPENDABLE CELLS OF THE BODY CAN BE REGARDED AS BELONGING TO CLONES WHICH HAVE ARISEN AS A RESULT OF SOMATIC MUTATION OR CONCEIVABLY OTHER INHERITABLE

CHANGES. Each such clone will have some individual characteristic and in a special sense will be subject to an evolutionary process of selective survival within the internal environment of the body.

IT IS BELIEVED THAT THE ADVANTAGES OF JERNE'S THEORY CAN BE RETAINED AND ITS DIFFICULTIES OVERCOME IF THE RECOGNITION OF FOREIGN PATTERN IS ASCRIBED TO CLONES OF LYMPHOCYTIC CELLS AND NOT TO CIRCULATING NATURAL ANTIBODY. The resulting formulation may be stated as follows:

The plasma-globulins comprise a wide variety of individually patterned molecules AND PROBABLY several types of physically distinct structure. Amongst them are molecules with reactive sites WHICH CAN CORRESPOND PROBABLY with varying degrees of precision to all, or virtually all, the antigenic determinants that occur in biological material other than that characteristic of the body itself. Each type of pattern is a specific product of a clone of mesenchymal cells and IT IS THE ESSENCE OF THE HYPOTHESIS THAT EACH CELL AUTOMATICALLY HAS AVAILABLE ON ITS SURFACE REPRESENTATIVE REACTIVE SITES EQUIVALENT TO THOSE OF THE GLOBULIN THEY PRODUCE. For the sake of ease of exposition these cells will be referred to as lymphocytes, it being understood that other mesenchymal types may also be involved. Under appropriate conditions, cells of most clones can either liberate soluble antibodies or give rise to descendant cells which can.

IT IS ASSUMED THAT WHEN AN ANTIGEN ENTERS THE BLOOD OR TISSUE FLUIDS IT WILL ATTACH TO THE SURFACE OF ANY LYMPHOCYTE CARRYING REACTIVE SITES WHICH CORRESPOND TO ONE OF ITS ANTIGENIC DETERMINANTS. The capacity of a circulating lymphocyte to pass to tissue sites and there to initiate proliferation is now relatively well established (cf. Gowens, 1957; Simonsen 1957). IT IS POSTULATED THAT WHEN ANTIGEN-NATURAL ANTIBODY CONTACT TAKES PLACE ON THE SURFACE OF A LYMPHOCYTE THE CELL IS ACTIVATED to settle in an appropriate tissue, spleen, lymph node or local inflammatory accumulation, AND THERE UNDERGO PROLIFERATION TO PRODUCE A VARIETY OF DESCENDANTS. In this way preferential proliferation will be initiated of all those clones whose reactive sites correspond to the antigenic determinants on the antigen used. The descendants will include plasmacytoid forms capable of active liberation of soluble antibody and lymphocytes which can fulfill the same functions as the parental forms. The net result will be a change in the composition of the globulin molecule population to give an excess of molecules capable of reacting with the antigen, in other words the serum will now take on the qualities of specific antibodies. The increase in the number of circulating lymphocytes of the clones concerned will also ensure that the response to a subsequent entry of the same antigen will be extensive and rapid, i.e. a secondary type immunological response will occur.

SUCH A POINT OF VIEW IS BASICALLY AN ATTEMPT TO APPLY THE CONCEPT OF POPULATION GENETICS TO THE CLONES OF MESENCHYMAL CELLS WITHIN THE BODY. It is clear that the internal environment involved is an exceedingly complex one and in all

probability many factors will impinge on clones of antibody-producing cells from that environment. It is equally certain that inheritable changes (at the clonal level) will occur as a result of somatic mutation OR OF THE STILL OBSCURE PROCESSES RESPONSIBLE FOR DIFFERENTIATION DURING DEVELOPMENT OF REGENERATION AND REPAIR.

IT WOULD BE INAPPROPRIATE TO ELABORATE THIS VIEW MUCH FURTHER IN A PRELIMINARY COMMUNICATION, but it should be immediately evident that it has highly relevant implications for the general function of the lymphocyte, for the fact that sensitization and homograft immunity reactions seem to be mediated by lymphocytes or other mesenchymal cells without liberation of classical antibody, and for recent findings of extremely rapid liberation of antibody from normal cells. A PRELIMINARY SURVEY OF A VARIETY OF PATHOLOGICAL CONDITIONS WHICH INVOLVE ANOMALOUS IMMUNE REACTIONS ALSO SUGGESTS THAT THIS CELLULAR APPROACH HAS GREATER RELEVANCE TO THE PROBLEMS THAN ANY OF THE OTHER HYPOTHESES. These aspects will be elaborated in a more extensive contribution now in preparation.

One aspect, however, should be mentioned. THE THEORY REQUIRES AT SOME STAGE IN EARLY EMBRYONIC DEVELOPMENT A GENETIC PROCESS FOR WHICH THERE IS NO AVAILABLE PRECEDENT. In some way we have to picture a "randomization" of the coding responsible for part of the specification of gamma globulin molecules, so that after several cell generations in early mesenchymal cells there are specifications in the genomes for virtually every variant that can exist as a gamma globulin molecule. This must then be followed by a phase in which the randomly developed specification is stabilized and transferred as such to descendant cells. At this stage, again following Jerne, any clones of cells which carry reactive sites corresponding to body determinants will be eliminated. The necrotic effect of the tuberculin on sensitized fibroblasts might be taken as a crude analogue of the process by which clones with unwanted reactivity can be eliminated in the late embryonic period with the concomitant development of immune tolerance.

THE HYPOTHESIS HAS MANY OF THE SAME IMPLICATIONS AS THE BURNET-FENNER AND THE JERNE THEORIES. Its chief advantage over the former is its relevance to the nature of normal antibodies including the red cell isoagglutinins and the SIMPLER INTERPRETATION of tolerance to potential antigens experienced in embryonic life. Its advantages over Jerne's theory are its capacity to cover homograft and related types of immunity as well as the production of classical antibody, AND TO ELIMINATE THE VERY UNLIKELY ASSUMPTION that entry of a globulin molecule into a cell will stimulate the cell to produce exact replicas of that globulin.

DESPITE THE SPECULATIVE CHARACTER OF MUCH OF THE DETAIL OF THIS MODIFICATION OF JERNE'S THEORY --WHICH MIGHT BE REFERRED TO AS THE "CLONAL SELECTION HYPOTHESIS" --IT HAS SO MANY IMPLICATIONS CALLING FOR EXPERIMENTAL INQUIRY that it has been thought justifiable to submit this preliminary account for publication."

DOI:10.3322/canjclin.26.2.119

In Summary:

-Burnet admits to 3 different theories on the formation of antibodies (excluding the Side Chain Theory for some reason)

-he states the Indirect and Natural Selection were devised to explain two phenomena that the Direct Theory could not explain

-it was considered inconceivable that there could be a mechanism within the body which could detect all foreign material and no explanation was ever devised in regards to this hypothesis

-Burnet credits Jerne's hypothesis for allowing him to create his own alternative take

-Jerne ASSUMED that among the globulin molecules were all the necessary patterns to attack any antigen

-Burnet states that if the basis of Jerne's theory is accepted, than most phenomena can be explained by the theory

-Burnet states that Talmage believed Jerne's theory to be an extension of Ehrlich's Side Chain Theory

-Talmage accepts as the best basis for the continued future evolution of antibody theory that the replicating elements be of cellular rather than extracellular nature

-Burnet states that the Clonal Theory is simply a recognition that the expendable cells of the body can be regarded as belonging to clones which arise due to either somatic mutation OR CONCEIVABLY other inheritable changes

-Burnet believes he can keep Jerne's basic theory but improve on it by stating that the recognition of foreign material is ascribed to lymphatic cells rather than to natural antibodies

-it is the essence of his HYPOTHESIS that each cell automatically has available on its surface representative reactive sites equivalent to those of the globulin they produce

-Burnet ASSUMES that when an antigen enters the blood/tissue fluids that it will attach to any lymphocyte carrying reactive sites which correspond to one of its antigenic determinants

-he postulates that when a antigen attaches to a lymphocyte it activates the cell to produce descendants

-Burnet states that his point of view is basically attempting to apply population genetics to the clones of mesenchymal cells in the body

-he believes inheritable changes occur either at the somatic level or by the yet obscure process of differentiation of repair/regeneration

-he states it would be inappropriate to elaborate on his views any further in a preliminary communication

-he says preliminary survey of a variety of pathological conditions which involve anomalous immune reactions SUGGESTS his cellular hypothesis has more relevance than any of the other hypotheses

-he mentions that his theory requires at some stage in early embryonic development a genetic process for which there is NO AVAILABLE PRECEDENT

-Burnet states that his hypothesis has many of the same advantages as those of Burnet-Fenner (his previous theory) and Jerne

-Burnet admits that much of his theory is SPECULATIVE and requires experimental inquiry

Thus we have yet another in a long line of theories about the formation/function of antibodies. Realize that these theories are not based on any direct observations on antibodies themselves but are a collection of indirect observations of many unrelated and often contradictory experiments. These scientists then attempt to put together the results they like and create an explanation for them while disregarding evidence/theories which do not fit their criteria. But as can be seen by the previous antibody theories, there is always a new theory that seemingly comes along to wipe out the old ones.

This can be seen from these highlights from "The Clonal Selection Theory: what it really is and why modern challenges are misplaced" by Arthur M. Silverstein:

"THE CLONAL SELECTION THEORY (CST) 2 of Macfarlane Burnet and David Talmage SEEMS TO BE UNDER FIRE CURRENTLY FROM SEVERAL DIRECTIONS. Irun Cohen 3, in considering the role of autoimmunity in the economy of the body, suggested that, “PROGRESS IN IMMUNOLOGY APPEARS TO HAVE RENDERED THE CLONAL SELECTION PARADIGM INCOMPLETE, IF NOT OBSOLETE; true it accounts for the importance of clonal activation, but IT FAILS TO ENCOMPASS, REQUIRE, OR EXPLAIN MOST OF THE SUBJECTS BEING STUDIED BY IMMUNOLOGISTS TODAY ...”. Polly Matzinger4 has gone even further: based upon her view

that the “DANGER THEORY” has negated Burnet’s classical view of self-nonself discrimination, SHE EXTENDS THIS TO SUGGEST THAT THE ENTIRE CLONAL SELECTION PARADIGM THAT HAS RULED IMMUNOLOGY FOR SOME 35 YEARS HAS BEEN OVERTHROWN."

"GIVEN THIS WIDE-OPEN THEORETICAL TERRAIN, IT IS NO WONDER THAT DEBATE CONTINUES on such questions as a “big bang” versus the continuous generation of diversity, the relative roles of central versus peripheral mechanisms of tolerance, the number and type of signals required for one or the other response 43, whether autoimmunity is dangerous or beneficial (Cohen3) and whether the immune apparatus evolved to recognize infectious pathogens (Cohn 44 and Janeway 45), “danger” (Matzinger 46) or, following Jerne 47, “self” (Coutinho 48 and Cohen 3)."

"In the end, however, we must not lose sight of the fact that the CST IS ONLY A THEORY OF HOW ANTIBODIES ARE FORMED, NOT A THEORY OF WHY THEY ARE FORMED."

DOI:10.1038/ni0902-793

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