Amboceptor

Microbiological, virological, bacteriological, immunological, medical, epidemiological, historical, anecdotal

Tag: blood

Vermicious Macrogametocytes

I keep having ideas for “picture gallery” posts, involving different papers’ illustrations of the same sort of thing. So far I’ve only done one (the cockroach feeding apparati) as they turn out to be a lot of work and you can never tell which papers are going to have illustrations and which ones aren’t.

In searching for pictures of encapsulated roundworm larvae in various organs of various animals, I determined that they all look sort of the same. And it’s not that interesting to compare, say, a picture of mouse muscle to a picture of raccoon or porcupine muscle. It’s all muscle. Also it may not be a good idea to extend this blog into worms and other complicated parasite species. Malaria is a parasite and also an infectious disease. But what about worms? So in the area of parasitology, I may want to stick to protozoans.

Leucocytozoons are single-celled organisms that infect birds, and are transmitted by the bugs known as blackflies. They were first observed in the 19th century in owl blood, by a zoologist named Danilewsky working in Kharkov, Ukraine. Danilewsky named them for their resemblance to white blood cells, though the exact genus “Leucocytozoon” was not applied until 1904, as detailed in this historical report by Lithuanian pedant and protozoan expert Gediminas Valkiūnas.

They seem to have life cycles similar to malaria, being fellow members of the phylum Apicomplexa. They go through many stages of life. Sporozoites are generated in the gut of an insect, and migrate to the salivary glands. The insect injects them into the blood of a vertebrate, and they go through several more stages, first in the liver and then in red blood cells. Then an insect takes a blood meal, the parasites end up in the insect’s gut, and they eventually make more sporozoites. It seems that Leucocytozoons are not as specific as malaria parasites, as they often infect white blood cells as well as RBCs, and the sporozoites of some species thrive in places other than the liver.

* * *

This figure shows the “macrogametocyte” stage (macro-gamete-ocyte) of the parasite life cycle. The macrogametocytes grow inside red blood cells, eventually filling and distorting the cells. This may be a pretty standard illustration. But it brought something else to mind.

From Fallis AM, Desser SS, and Khan RA (1974), On species of Leucytozoon. Advances in Parasitology 12:1-67 : (available from the publisher, subscription required, or available in partial form from Google Books)

leucocytozoons-1974

As rendered by the Canadian authors of the paper linked above, these jaunty, big-eyed, dancing blobs are quite evocative. The genus ranges from L. vandenbrandeni (#2), which looks like a baleful ocean sunfish and lives inside the similarly aquatic birds called cormorants, to L. bonasae (#19), which infects grouse and appears to be wearing one of those baggy vinyl baseball caps from the golden age of breakdancing.

The Leucocytozoons bring to mind another type of sinister creature on a more macroscopic scale, from one of my favorite children’s books.

vermicious-knids-3

Yes, the Vermicious Knids, which menace visitors to the Space Hotel in Roald Dahl’s Chocolate Factory sequel, Charlie and the Great Glass Elevator. Described initially as resembling eggs with no features other than eyes, they then show that they could change shape.

"They're tremendously proud of being able to write like that." "But why say scram when they wanted to catch us and eat us?" "It's the only word they know,"

“They’re tremendously proud of being able to write like that.” “But why say scram when they wanted to catch us and eat us?” “It’s the only word they know.”

The macrogametocytes of Leucytozoons look especially similar to Vermicious Knids when the latter are only displaying one eye, as in the above message to humanity, or in these creative illustrations by comics artist Isaac Cates.

Noncanonical Vermicious Knids

Noncanonical Vermicious Knids

* * *

As you’d expect, most useful photos of these organisms are in color and therefore don’t look much like these ink drawings.

But here’s some beautiful drawings, in color, of red blood cells containing a similar parasite, Haemoproteus syrnii. Just like the first Leucocytozoons ever observed, this organism infects owls. What are they trying to say? Are they communicating by Braille or some other pattern-based system? Or by spelling out letters? They seem only capable of “C” and “O”.

From Karadjian G et al. (2013), Haemoproteus syrnii in Strix aluco from France: Morphology, stages of sporogony in a hippoboscid fly, molecular characterization and discussion on the identification of Haemoproteus species. Parasite 20:32 (11 pages) :

haemoproteus-syrnii

The paper (from folks at France’s National Museum of Natural History) is free.

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What’s a “full-sized drop” in nanoliters?

A lot has been made nowadays about reproducibility, and how for some reason the most interesting scientific results are often the least reproducible. Evidently when there’s huge pressure to generate a certain graph containing certain data, either to give yourself a chance at fame and fortune or to give yourself a chance to have your lab and scientific career continue to exist, sometimes the graph does not represent the sort of objective reality that exists throughout space and time. This can be because of wishful thinking, because of selective use of the data that seems most solid, because of variables we never considered which later turn out to be crucial, who knows what else.

There have always been experiments where different labs get different results. Often we say that “in our hands,” we get Result X, but another laboratory gets Result Y, and we can say this without accusing anyone of malfeasance. It’s acceptable to get different results.

But nowadays, a scientist has no excuse when his contemporaries can’t even figure out how to replicate an experiment. A hundred years ago, it was more tricky. Even if you wrote and asked for a detailed protocol, it would probably involve terms that had no exact meaning. The topic under discussion today is the word “drop”.

* * *

bass-watkins-test

In the December 1910 issue of the JAMA organ Archives of Internal Medicine, Drs. C. C. Bass and John A. Watkins wrote up (1) a new quick-and-easy test for typhoid that they had developed in the laboratories of Tulane University. The abstract is here, with subscription required to read the article. But this volume is old enough that it’s out of copyright and should be in Google Books. Though to be honest I can’t find it there and only found it at archive.org.

bass-watkins-figure1

Over the ensuing years many doctors’ offices used it with more or less success, but as you might expect, many found they were unable to get results and went back to their old routine of sending samples to a clinical lab and waiting a day or two.Even though Bass and Watkins went to the trouble of including highly mundane photographs of things like proper slide-rocking procedure, people couldn’t figure out what exactly they were supposed to do. The text still contains phrases like “two to three drops of an equal number of bacilli units and agglutinin units sufficiently dilute to prevent rapid agglutination”. And “this one-quarter drop of blood is about the quantity we use in making blood slides in examinations for malaria, differential counts, etc.” The instructions are easy to understand, but really to communicate this sort of information you have to show people and let them practice it.

As a result, the 1910s saw some skeptical questions came in to the miscellaneous letters section of JAMA, sometimes with fairly impatient replies from the editors.

von-pirquet-reaction-1914

The editors of JAMA explain in detail the benefits of the Bass-Watkins test. (citation #2)

Following the usual routine of randomly skimming randomly selected old journals, I found a follow-up piece in the July 1918 New Orleans Medical and Surgical Journal (3) which goes into further detail trying to delineate concepts like “drop”. The author (presenter, rather, since this is the transcript of a talk which was followed by comments) is one of Bass and Watkins’s junior colleagues at Tulane, Foster M. Johns. Here’s what he says.

In the eight years that have elapsed since the publication of this article, this reaction has constantly grown in favor of the clinicians of the South, in spite of many improper lots of reagent supplied by private laboratories, my own included, as well as the various biological houses. During this time the test has been in constant use in the laboratories of clinical medicine with which I am connected, and it is with the belief that this reaction offers an easier, quicker and even more accurate reaction to not only the clinicians, but the trained laboratory worker as well, that I have prepared this discussion of a well-known test. During this time the few faults in technic and production brought out by continual use have been met and overcome, with the exception of a technic that will insure the uniform production of a stock suspension of typhoid bacilli that will keep well under the ordinary conventions of usage.

As simple as the technic sounds, there is often considerable difficulty in doing a simple thing. Taking up the test step by step, I will endeavor to point out the places where error may creep in. To begin with, an absolutely clean slide, freshly washed with soap and water to remove the grease and dust, most be used. Now, we require one-quarter of a drop of blood on the center of the slide. This is a quantity almost impossible to describe to one not accustomed to the routine making of proper blood smears, but practically it is easily approximated. Squeeze a quantity of blood out of a puncture on the finger or ear lobe that will not quite drop off, and then barely touch the slide to it. The quantity adhering to the slide will vary from one-quarter up to one-half of one drop. In either instance, for practical purposes, the end result will not be influenced… The actual dilution of the organisms will not be disturbed by either of the quantities of blood, as the blood is then spread roughly over the middle third of the slide and allowed to dry.

Now, one drop of plain water is added. Drops can vary enormously in size, and while, if the proportions in the test were carried out to suit, no harm would ensue, still, for working purposes, we need a full-sized drop. In this instance the standard drop is measured by preferably using the ordinary medicine dropper held almost parallel to the table, so that the drop collects on the side of the elongated glass tip of the dropper.

Full-sized drop? Quarter drop? A quarter drop is not a drop divided into fourths, but a drop that will not quite drop off? Or do you really mean a “finger or ear lobe that will not quite drop off”? In which case the real concern may be not typhoid, but leprosy.

Instead of all this … wouldn’t it be easier to measure volume in microliters?

I know nothing about the history of scientific equipment, but Wikipedia reports that there were no micropipettes until 1960.  I don’t think there were syringes capable of measuring volumes on the level of a drop, either. And if there were such devices, they were far from disposable, and would need to be cleaned and dried between uses.

* * *

What is a drop anyway? And what was the smallest amount of volume that could be accurately measured a hundred years ago?

Again to the Wikipedia, which claims that today there is a medical definition of “drop” as 50 microliters. That means a quarter drop is 12.5 microliters, which sounds like about the right amount for a blood smear that you would quickly look at under a microscope.

Apothecaries traditionally were able to make much more precise measurements of weight than of volume. The common measurement of a “grain” is equal to 1/20 of a scruple, or 1/60 of a dram. And a dram is only 1/8 of an ounce, so there are 480 grains in an ounce, making a grain about 64 milligrams, under the old system where there were 12 ounces in a pound and a pound was about 1/3 of today’s kilogram.

Meanwhile, for liquids, people didn’t have too much trouble measuring in terms of scruples (slightly more than a cc or milliliter). But the minim, the volumetric equivalent of the grain, was only invented around the beginning of the 19th century, and required quite specialized equipment. Being the equivalent of a grain, the minim is about 64 microliters, or roughly… a drop. So it just made sense to refer to things in terms of drops. But when you factor in surface tension, temperature, the size of the vessel from which the drop is dropping… it’s always a judgment call. As the old saying goes, blood has a higher viscosity and specific density than water.

So those of us with access to space-age technology like micropipettes (and disposable anything) should count our blessings.

* * *

1. Bass CC, Watkins AA (1910). A quick macroscopic typhoid agglutination test. Arch Inter Med VI(6):717-729.

2. from “Miscellany”, September 5th (1914). Value of von Pirquet reaction in adults / Reliability of Bass-Watkins test. J Am Med Assoc LXIII(10):883.

3. Johns FM (1918). The Bass-Watkins agglutination test for typhoid. New Orleans Med Surg J LXXI(1):22-27.

Two Octaves down

From looking up the basics of complement fixation, it turns out that it had other names. It was called complement deviation: instead of complement landing on cells and lysing them, its path was deviated to some other end. Just like complement fixation: before it can land on cells and lyse them, complement is fixed in place and stays in solution. This is all very straightforward.

It was also called “The Bordet-Gengou Reaction”. That sounds more interesting.

* * *

The best source for info on the early history of immunology is the book Immunology: Pasteur’s Heritage, edited by Pierre-André Cazenave and published in 1991 by Wiley. I have not read this book, but I will bet you $12 that it’s the best source for info on the early history of immunology. The passages available in Google Books are great.

Anyway, according to Wikipedia, complement itself was first called “alexin“, around 1890 by researcher Hans Ernst August Buchner, who observed that serum contained a substance that could lyse bacteria in the absence of cells. Richard Pfeiffer observed a similar phenomenon. Also around 1890, the word “antibody” (technically “Antikörper” because they were all publishing in German) was coined, by Paul Ehrlich whose laboratory was more focused on the interactions of individual molecules (in this case antibodies, which he saw could neutralize the harmful effects of toxins) than cells. All of these researchers were sort of rivals to the organization of Élie Metchnikoff at the Institut Pasteur in Paris, which was focused on the newly discovered phagocytic cells that he thought were responsible for all aspects of immune protection, more or less.

Jules Bordet

Jules Bordet

Belgian researcher Jules Bordet was an apprentice to Metchnikoff. And Octave Gengou was Bordet’s brother-in-law and close colleague. At some point in the late 1890s (I can’t tell which paper it was; all their papers were in Annales de l’Institut Pasteur, and most of them are just called “Recherches sur la coagulation du sang” with vague subtitles), Bordet showed that “alexine” was made up of two parts: a heat-stable part, which just bound up things like toxins and bacteria; and a part that could easily be destroyed by heat. Without this “heat-labile” element, the bacteria didn’t get lysed.

This led the Ehrlich school to divide its “Antikörper” concept into two parts: Amboceptor (heat-stable), and Komplement (heat-labile). Meanwhile the Bordet/Metchnikoff school divided it into substance sensibilitrice (sensitizing substance) and alexine (bacteria-lysing substance). Bordet showed that Amboceptor/sensibilitrice/antibody against a certain substance would only be present in the blood of people immunized with that substance; while just about any blood contained Komplement/alexine/complement which could lyse anything.

* * *

What about Gengou?

It looks like Bordet and Gengou start being paired together on these papers in 1901, which was also the year that Bordet was invited to be the director of the newly founded Institut Pasteur. Not the real one… the Institut Pasteur du Brabant in Brussels. Which was the only “Institut Pasteur” not to be an offshoot of the original one in France; this one was just called that because Madame Pasteur gave it her blessing. In the 1980s it moved away from the beautiful building on Parc Léopold, which now houses the Bavarian Representation to the EU (pdf), and was eventually merged with the Belgian government’s Scientific Institute of Public Health.

Jules Bordet and Octave Gengou are credited with discovering complement fixation. Earlier they had seen that the amboceptor/complement mixture that destroys bacteria (bacteriolysis) can also destroy red blood cells (hemolysis), provided the RBCs were from a different species or a different blood group (yes, they basically also discovered blood groups).

But if you take some serum with a lot of amboceptor and complement in it, and do a bacteriolysis… all the complement should get used up. Then you mix it with sheep red blood cells, and they don’t get lysed. What you just did was prove that the serum contained antibody against that particular bacteria. You can see how this would be useful. This is the Bordet-Gengou reaction.

* * *

What about the Nobel Prize?

Bordet and Gengou also discovered the bacterium that causes pertussis, which is now called Bordetella. That looks less silly than Gengouella, but the latter actually sounds better, to the cultured and discriminating ear, They also developed Bordet-Gengou agar, the ideal substance for growing Bordetella pertussis and distinguishing it from other small Gram-negative coccobacilli, In 1919, the first Nobel Prize ceremony in five years was held, and the prize in medicine and physiology was given to Bordet, aged 49, for achievements summarized most briefly as:

  • Complement
  • Complement fixation
  • Pertussis bacterium

Two of those are accomplishments shared with Gengou. And both of those were procedures named after both Bordet and Gengou: in addition to the Bordet-Gengou complement phenomenon, the isolation of Bordetella pertussis depended on culturing it on what is known as Bordet-Gengou agar). And yet, only one person got the prize.

octave-gengar

Octave Gengar

Granted, the prize had always been given to a single person: with two exceptions. One of them was in 1908, when it went to both Pasteur and Metchnikoff. Not to do that might have caused an outbreak of bad feelings to rival the Schism of 1054. I suppose Gengou would not mind the solitary recognition of his great collaborator.

Really, the biggest injustice is not the lack of a Nobel Prize, or the zero Google hits for the word “Gengouella”, but my inability to find any photos of him online.

* * *

The name “Octave” rang a bell. I don’t know if I’d heard of anyone by that name until this year. Who was the other one? Quickly the memory became clear: Octave Chanute.

octave-chanute

Octave Chanute

From flyingmachines.org:

Octave Alexandre Chanut*  , a native of France, was retired from a distinguished engineering career and living in Chicago, Illinois, when he began to pursue his life-long interest in aeronautics. His experiments with “gliding machines” began in 1896 and were conducted at Miller Beach and Dune Park, Indiana, on the southern shore of Lake Michigan.

Octave Chanute went on to be the main enthusiast for the Wright Brothers during their early aerial trials, encouraging them and supplying them with the latest aerial information. By 1900 Chanute had become the center point for various aerial experimenters in Europe and the U.S. His 1894 book “Progress in Flying Machines” was a landmark volume and was the book recommended to Wilbur Wright by the Smithsonian Institution in 1899.

One of the messages I got from the displays at the Wright Brothers National Memorial, on North Carolina’s Outer Banks, was that the brothers cared deeply about impressing Octave Chanute, whose correspondence served as a nexus for information exchange between different experimentalists in heavier-than-air flight. Chanute was unquestionably the nearest “giant” on whose shoulders they stood. But I had never heard of him until this year. Maybe you had.

Takeaway message: If your name is “Octave”, “Staff”, “Rest”, “Flat” or another musical term, your contributions may be overlooked by the eye of history. You may want to change your name. Make a note of it. But don’t make “Note” your name.

* * *

Sources:

What is an amboceptor?

Amboceptors get their name…

Or I could say “Amboceptor gets its name…” — it’s one of those words that can be an object, or a substance.

Amboceptor gets its name from the Latin root “ambi-“, because it’s a receptor for two things.

An amboceptor is something that attaches to an antigen, and then attaches to complement molecules. When this happens on a cell surface, the complement pokes holes in the cell to damage it. When it happens in solution, it blocks the complement from attaching to cells.

Isn’t that… an antibody? An amboceptor is an antibody.

Yes, but we didn’t always know that.

* * *

We knew there were amboceptors. We knew there were antibodies. There could have been amboceptors that weren’t antibodies.

Amboceptor, as a substance, is critical in the complement fixation assay for serum antibody. This is a once-ubiquitous, now-too-complicated-to-be-worth-doing process of diagnosing diseases like syphilis by looking for (anti-syphilis) antibodies in a patient’s serum. It was a fixture in American life, under the monicker of the “Wassermann Test”, which countless practitioners trusted to confirm a diagnosis of syphilis, or even to diagnose syphilis in the absence of other evidence. You know how nowadays we have urban legends about babies named things like “Urine” and “Female” based on hospital mix-ups and parental idiocy? That list used to include “Positive Wassermann Johnson”.

How real?

How real?

Probably the best description of complement fixation on the internet is provided here by an organization that is always at the vanguard of science and health policy, the Texas state government. I’ll try to summarize in terms that would be familiar to blog readers from 1920.

  • Sheep corpuscles (red blood cells) are mixed with amboceptor. This sensitizes them to the complement.
  • If the sensitized corpuscles are then mixed with complement, the complement binds them and starts poking holes in the corpuscles, which is called “lysing” them.
  • The solution turns pink as the contents of corpuscles spill out. This is called “laking”. Or it was back then, anyway.
  • Serum is a solution containing amboceptor as well as many other things. If you take serum from a syphilitic patient, and mix it with syphilis antigen, the amboceptor should bind the antigen and form a complex in solution.
  • If you mix this with complement, the antigen-amboceptor complexes should bind up the complement before it can attach to the corpuscles and lyse them.
  • If you take serum from a non-syphilitic patient, the amboceptor will NOT bind the syphilis antigen, and therefore it will not bind the complement. The complement will not bind to amboceptor alone, only to complexes. So the complement is still available for lysing and laking.

As you can imagine, this is a touchy procedure prone to error.

First of all, what about the complement that’s already in the human serum? You have to inactivate it, so you can then see how the rest of the serum reacts to externally introduced complement that you bought from a supplier. How do you inactivate it? Complement is protein and you heat it to denature it. You heat it at 56 degrees Celsius. Exactly 56 degrees Celsius. For exactly 30 minutes. At 54 degrees, you won’t denature enough complement. At 58 degrees, you’ll also denature antibodies. If you do it for 60 minutes, you’ll also denature antibodies.

But that’s assuming your serum was isolated from the blood in a timely fashion, and kept at room temperature for 24 hours or less. Or maybe it’ll work after 48 hours. What if it’s been refrigerated? That has some effect on the protein stability, maybe they will start to aggregate and become useless. What if it hasn’t sat at room temperature for long enough? Blood needs to sit for an hour or so before you start processing it to remove the corpuscles and platelets, after all. How perfect does the procedure need to be? All you are showing is that there is something in the serum that blocks complement from depositing on the sheep cells. What if that something isn’t amboceptor?

And what if the serum contains something that’ll lyse the sheep corpuscles on its own? After all, sheep are not human and some people may have a natural antisheep hemolysin, which is a word used for amboceptors or other substances that lyse corpuscles. How do you correct for that? Well, you have to include a control with no complement, to figure out the background amount of lysing that goes on. Then you can dilute the serum properly. But if you dilute the serum, you make the procedure less sensitive. Some laboratories will do this, some will not.

And how much complement should you add to the serum? If you add too much, you get false negatives. Someone with early-stage syphilis may have a low level of amboceptor. If there’s too much complement, you’ll soak up all the amboceptor and still have plenty of complement left over. But it’s possible to add too little complement, too. With too little, you get ambiguous cases, where you’re not sure if you see laking or not. What if you don’t see laking in any of your samples? You need a positive control. Does that mean you need to have fresh serum on hand at all times from a known syphilitic? Oh dear.

And what should the antigen be, that can pick out syphilis amboceptor from all the other amboceptor in the serum? There’s no way of growing the syphilis spirochetes in your laboratory. Extract of syphilitic liver was used originally by Wassermann himself. Usually you would exploit the unusual lipids found in the spirochetes, by using some uninfected tissue, like some sort of heart or liver extract, with added cholesterin (a.k.a. cholesterol). I prefer Noguchi’s acetone-insoluble fraction of the alcoholic extract of fresh beef heart, or as I call it, NAIFAEFBH. In 1941 Pangborn will revolutionize syphilis diagnosis by standardizing this antigen as a certain combination of cardiolipin, cholesterol, and lecithin, all taken from beef heart. This will still be used in the 21st century [Castro et al. (2000), Clin Diagn Lab Immunol 7(4): 658-661].

And what about the Hecht-Weinberg modification, particularly the procedure of Gradwohl? A lot of people are not comfortable with adding external complement to the serum, instead trusting that the natural amount of complement in the serum is appropriate for the process. This means you wouldn’t heat-inactivate at all. This means other controls will have to be done. And the complement may end up getting inactivated anyway, based on natural half-life, exposure to light, etc.

* * *

And on, and on, and on. Most of the contemporary articles about these issues are 100% baffling to a 21st-century reader, but one of the least baffling is an exchange in the November and December 1914 issues of The Lancet-Clinic. No, not The Lancet. The Lancet-Clinic, which was published in Cincinnati until going out of business in 1916. Unfortunately it isn’t indexed in Pubmed or Google Scholar, but random Google searches for “Wassermann test” and “amboceptor” quickly found it in Googlle Books.

Go to your bookshelf, the one that groans under the weight of bound volumes of turn-of-the-century Ohio medical journals. Open up The Lancet-Clinic, Vol. CXII, issue 26, and turn to pages 536-539, where Albert Faller, M.D. issues forth a torrent of verbiage on all the problems he sees with the Wassermann test, and finishes by extolling the virtues of Gradwohl’s Hecht-Weinberg modification, which turns many negative results into positives. This is then followed by harrumphing from one Dr. Berghausen, who sees no reason why heating would ever cause any harm to anything, and opines that physicians should stop being afraid of reporting negative results just because they happen to conflict with the fact that a patient seems to have syphilis. Dr. E.A. North then muses that perhaps all this could be cleared up if people would mix their samples thoroughly, and Faller rounds out the exchange by saying that far from being “tired of condemning individuals as sufferers of syphilis”, he simply does not trust that a patient who goes from positive Wassermann to negative Wassermann has actually been cured. And he does trust when that happens with the Hecht-Weinberg. Finally, in the December issue [Lancet-Clinic CXII(27): 624], the great man himself, Rutherford Birchard Hayes Gradwohl, writes in to further heap scorn on the deplorable Berghausen, saying that “I wish to state as one who has been instrumental in urging this test before the American profession, no such ambitions have ever surged through my serological breast.”

* * *

The Wassermann test was a serious concern. For decades, in most U.S. states, people were required to be tested for various diseases, particularly STIs, particularly syphilis. Here in Pennsylvania we waited until 1997 to repeal the requirement, which had begun as part of a nationwide campaign by New Deal-era Surgeon General and future Pittsburgh public health supremo Thomas Parran. But what does it mean to have a negative Wassermann?

What if you have no signs of the disease, you have no family history of it, but you have a positive Wassermann? Does it mean anything? How can you not worry? What if I had a positive reading earlier, and now I’m negative? Am I cured, thanks to the new sulfonamide drugs that work so much better than salvarsan? Is it really possible to be cured? And doesn’t this just measure antibodies? I have antibodies against just about every infection I’ve ever suffered. That doesn’t mean I’m still an infection risk.

How to get reliable Wassermann results was a perennial topic in the literature for decades. Because it was such a devastating diagnosis. And for practical reasons, since so many of the tests had to be carried out and they were so cumbersome.

A sample:

  • Noguchi, H. and J. Bronfenbrenner (1911). The Comparative Merits of Various Complements and Amboceptors in the Serum Diagnosis of Syphilis. J Exp Med 13(1): 78-91.
  • Van Saun, A. I. and M. K. Preston (1918). Comparative Wassermann Tests with Two Antigens. Am J Public Health (NY) 8(2): 146-148.
  • Lewis, P. A. and H. S. Newcomer (1919). Observations on the Wassermann Reaction: A Comparison of the New System of Noguchi with That Using Cholesterolized Antigen According to McIntosh and Fildes. J Exp Med 29(4): 351-359.
  • Browning, C. H. and E. L. Kennaway (1920). Suggestions for a New Criterion of a Positive Wassermann Reaction Based on an Analysis of 2334 Quantitative Tests. J Hyg (Lond) 19(1): 87-106.
  • Mackie, T. J. and C. C. Rowland (1920). The Value of Simultaneous Testing for the Wassermann Reaction, with Two Different Antigens and the “Ice-Box Method”. Br J Exp Pathol 1(5): 219-224.
  • D.Aunoy, R. (1921). Comparative Study of the Wassermann and Sachs-Georgi Reactions. J Med Res 42(4): 339-347.
  • Bigger, J. W. (1921). The Reliability of the Wassermann Test as performed by different Pathologists. J Hyg (Lond) 20(4): 383-389.
  • Famulener, L. W. and J. A. W. Hewitt (1922). Studies on the Serodiagnosis of Syphilis: I. The Hecht-Weinberg-Gradwohl Test. J Inf Dis 31(3): 285-290.
  • Shepardson, R. T. (1922). Preliminary Report on an Investigation of the “Provocative Wassermann” Controlled by the Ice-Box Method. Cal State Med J 20(3): 80-83.
  • Dulaney, A. D. (1923). The Wassermann and Kahn Precipitation Tests Compared in 900 Cases. Am J Public Health (NY) 13(6): 472-474.
  • Osmond, T. E. and D. McClean (1924). A Comparison of the Kahn and Wassermann Tests on 500 Serums. Br Med J 1(3301): 617-618.
  • Ruediger, E. H. (1924). A Plea in Favor of a Standardized Wassermann Test. Cal West Med 22(11): 548-553.
  • Malcolm, M. (1924). A Comparison of the Kahn Test with the Wassermann Test. Can Med Assoc J 14(3): 222-224.
  • Wyler, E. J. (1927). A Note on Two Factors Affecting the Sero-diagnosis of Syphilis. Br J Vener Dis 3(4): 320-325.
  • Green, F. (1929). The Kahn Test as Compared with a Standard Wassermann Reaction. Can Med Assoc J 20(1): 26-29.
  • Ferguson, J. H. and E. C. Greenfield (1929). Value of the Hinton Test in the Serum Diagnosis of Syphilis: In Comparison with the Khan and Wassermann Tests. Br Med J 1(3558): 492-494.
  • Evans, N. (1930) Kahn Precipitation Test for Syphilis: As Used in Conjunction with the Wassermann Test. Cal West Med 32(1): 24-26.
  • Eagle, H. (1931). Studies in the Serology of Syphilis: IV. A More Sensitive Reaction for Use in the Wassermann Reaction. J Exp Med 53(5): 605-614.
  • Chambers, S. O. (1932). The Kahn Precipitation Test: Compared with the Kolmer Modification of the Wassermann Test in Untreated Primary Darkfield Positive Seronegative Syphilis. Cal West Med 37(3): 153-155.
  • Barritt, M. M. and A. O. Ross (1939). A Comparison of the Wassermann and Meinicke (M.K.R. II) tests in the Serological Diagnosis of Syphilis. Br J Vener Dis 15(3): 183-202.
  • Richardson, G. M. (1940). The Specificity of the Bordet-Wassermann Reaction: Preliminary Note on an Improved Method. Br J Vener Dis 16(3-4): 166-185.
  • Rickword Lane, C. (1944). Comparison of the Laughlen Reaction for Syphilis with the Wassermann and Kahn Reactions. Br J Vener Dis 20(2): 78-81.
  • Kolmer, J. A. (1944). The Problem of Falsely Doubtful and Positive Reactions in the Serology of Syphilis. Am J Public Health Nations Health 34(5): 510-525.
  • McMenemey, W. H. and W. H. Whitehead (1949). Ford Robertson and Colquhoun Modification of the Meinicke Clarification Reaction Compared with the Harrison-Wyler Wassermann and the Standard Kahn Reactions. Br J Vener Dis 25(3): 147-154.
  • Osmond, T. E. (1950). Comparison of the Wassermann and Kahn Reactions. Br Med J 1(4652): 524.
  • Bekierkunst, A. and F. Milgrom (1950). Complement-fixation Reactions with Cardiolipin Antigen Compared with Kahn Reactions. Bull World Health Organ 2(4): 687-688.
  • Orpwood Price, I. N. and A. E. Wilkinson (1952). Comparative Serum Testing with Cardiolipin and Crude Heart Extract Wassermann Antigens. Br J Vener Dis 28(1): 16-19.
  • Kahn, R. L. (1972). Syphilis Serology with Lipoidal Antigen: The Meaning of Positive Reactions. J Natl Med Assoc 64(2): 117-passim.

In addition to any number of more technical papers with titles like “The Amount of Hemolysin Absorbed By Sheep Corpuscles”, published in the American Journal of Syphilis.

The situation was summed up well by Knox College art benefactor L. W. Famulener and Julia A. W. Hewitt (1922), in the atypically lucid introduction to their typically inconclusive study.

It is quite difficult to delimit the borderline between the nonsyphilitic and the syphilitic person. Supersensitive laboratory tests on nonsyphilitic serums* in certain cases may give results which pass over to the syphilitic side. This is especially true when the worker is in pursuit of those who clearly show syphilitic conditions, but whose serums fail to give a positive reaction with the usual methods. The nonreacting syphilitic may not even carry “syphilitic fixing bodies” in his serum. It is well known that there is a decided quantitative difference existing between the different positive serums. If these “fixing bodies” are metabolites, they may arise anew, or may be normal substance markedly increased in amount during the course of the disease. In the latter case, it is quite conceivable that certain nonsyphilitic persons may naturally have an abnormally large content of these substances in their blood, while, on the contrary, known syphilitic persons may fail to elaborate these bodies and show only a very low content, even below the average norm for the healthy person. Therefore memers of either group may be found on opposite sides of an arbitrarily established borderline, where clinically they do not belong. …

As to the criteria which should determine the syphilitic from the nonsyphilitic persons, no common agreement exists. By cooperative studies among clinicians, pathologists and serologists, progress may be made toward that ultimate end. In the absence of a standard method for the serum diagnosis of syphilis, a multitude of modifications of the original Wassermann technic** have come into existence. Many of these modifications are erroneous in conception, even conflicting with the established laws of serology, consequently leading to false results.

And the number of modifications would only increase, until the complement-fixation process was given up as inherently unstable and imprecise, the flintlock musket of the serologist’s arsenal.

* Nowadays we say “sera”. We must be more pretentious nowadays.

** Nowadays we spell it “technique”. Pretentious?

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So anyway. Why don’t we use the word “amboceptor” anymore? Is it still used in some circles? No, I don’t think so. A Pubmed search for “amboceptor” [All Fields] found 59 results. The last one was in 1991, and they were distributed fairly evenly over the previous 87 years. Which is pretty amazing when you consider how many more articles are indexed from 1991 than 1904. And in fact, most of the results since 1965 are either translated out of German, Czech, or Bulgarian, or are in odd journals like Developments in Biological Standardization and Bibliotheca Haematologica. The last appearance in an abstract is in a German-language paper entitled “Effect of different media on long-term cultivation of human synovial macrophages”, from the now-retired Eckhard Stofft of the University of Mainz, a specialist in tendon and ligament pathology.

Scientometric metadata

Scientometric metadata

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“In the extensive experiments and observations during the past twenty years, the greater amount of effort has been expended in the consideration of the manner of the production of immune bodies upon the part of the host, together with quantitative and qualitative methods for the determination of such bodies. Thus specific substances have been recognized, to which the names antitoxin, agglutinin, precipitin, amboceptor, opsonin, etc., have been given.”

– Duval, C.W. and F.B. Gurd (1911). Experimental Immunity with Reference to the Bacillus of Leprosy. Part I: A Study of the Factors Determining Infection in Animals. J Exp Med 14(2): 181-195.

Antitoxin: still used. Although all antitoxins are antibodies, the word is still medically useful.

Agglutinin: still used, but usually we say “hemagglutinin”. Not all hemagglutinins are antibodies.

Precipitin: still used. Although all precipitins are antibodies, the precipitin test, which is even older than the complement fixation test, is still used.

Opsonin: still used. A very useful word. And not all opsonins are antibodies.

Amboceptor: obsolete.