RH Negative Blood: Blood Feud—Which Scientist Discovered the Rh Factor?
For simplicity, RhD is used for Rh positive blood and Rhd is used for Rh negative blood in this series of blood articles.
You think you know who discovered the Rh Factor, right? I guarantee to introduce you to new facts in this article. What you have to ask yourself is this: Is the Rh Factor a blood test or is the Rh Factor a genetic disease? Let me know if this article changes your mind.
In the last article, we’ve reviewed the lab test procedure, using rabbit anti-rhesus sera, and how doctors, Landsteiner and Wiener in 1937, discovered a weak but new X Factor (blood protein). They were unimpressed by their finding and didn’t share the discovery with their peers. So, the next question is how did this simple, unnamed lab test get involved in the discussion of an Rhd mother pregnant with an RhD child? Yeah? How did it?
In simple terms, a murky, weak blood test has brought most of the confusion to the topic. The relationship of the blood test to disease goes unexplained or when it is, laymen don’t understand. The first key fact is: the rabbit anti-rhesus sera identified the people with what became known as Rh negative blood two years later. The men had found another X Factor but they didn’t share or publish their discovery. They put the test away. Landsteiner and Wiener went back to their test after a colleague, Levine, published his paper in 1939. This paper was the first shot fired in what became known as a “blood feud” between Levine and Wiener. This is what happened.
Blood transfusions took off after refrigeration and anticoagulation (treating blood with chemical sodium citrate which prevents it from clotting). Because of refrigeration and anticoagulation, donor blood could be collected and kept on hand for transfusion. Hospital blood banks were established and then transfusions became available. Before this the donor had to be brought to the recipient, and the blood transfusion would take place arm to arm. A rise in transfusion reactions occurred. Yes, blood was given for pick-me-ups and minor anemia. Rhd women got sensitized and developed antibodies without getting pregnant. (1) Zimmerman. In short, doctors abused the notion of transfusing blood and created future problems for Rhd women. In 2013, we don’t get transfusion pick me ups, and we don’t get a transfusion without blood tests.
However, doctors also noticed people didn’t have visible reactions the first time they were transfused with blood, but did on the second round. Their conclusion: people were sensitized by some unknown X Factor. The same factor(s) Landsteiner had pursued. Remember, Landsteiner and Wiener didn’t write a paper about their weak blood test with the rabbit anti- rhesus sera, the test hadn’t been named yet, and at the time of their discovery they didn’t find it important.
In 1937, Doctor Phillip Levine was called in to consult on a case at Bellevue Hospital. A woman gave birth to a macerated fetus, and she badly needed a transfusion. She was given 500 cc of her husband’s type O blood which matched her blood type. She had a severe reaction and started bleeding again. Lab technicians were confused, and her blood was retested. She remained type O like her husband, and according to the known science at the time, she shouldn’t be having this type of reaction. Rh negative blood hadn’t been discovered in live patients yet.
Levine had previously worked in Landsteiner’s lab and knew how to make test reagents. So, Levine made a test serum from her blood and tested it against 104 samples of type O blood in the blood bank and found only 21 compatible to her. The patient was transfused with a compatible match and lived. (2)
Levine’s procedure: He used the woman’s own blood as a test serum against all available type O blood donors at the hospital. He got the same clumping reaction with her blood that Landsteiner and Wiener did with the rabbit anti-rhesus sera. Only Levine didn’t know about their test because they hadn’t published it or given talks on their new discovery. According to Zimmerman, Levine published his paper a year later after wrestling over the ethics of breaking his pledge to Landsteiner. Levine said he published to benefit science and end suffering. (1)
Levine was surprised by the result after testing revealed the woman’s reaction didn’t have anything to do with the blood factors M, N, and P that he had discovered with Landsteiner. Again, Landsteiner and Wiener were unimpressed with their “weak test” with the rabbit anti-rhesus sera, and they didn’t write a paper on it. After Levine and Stetson published a case study on this woman in 1939, Wiener and Landsteiner rushed out a paper six months later in 1940. In his paper, Wiener gave the test a fancy name, “Rh Factor.” Wiener worked with monkeys and apes and published frequently on his findings. (4)
Levine, perhaps because he had broken his promise to Landsteiner not to do blood research, didn’t name his test. If he had, the argument we now see on the internet about pure blood separate and free of monkey/ape blood would have never entered the Rh negative blood story. Levine didn’t name his test, Wiener did, and the Rh Factor stuck. (5)
Note to Readers: Two things: Levine made his own reagent with the woman’s blood and discovered an X Factor that caused a transfusion reaction. Plus, he saved her life. I don’t know why he let Landsteiner and Wiener horn in, but as time went on he tried to make his contribution clear. Second: The pledge Landsteiner asked Levine to follow when he left the lab “no blood work research” was a gentleman’s agreement and not a contract. Such pledges were accepted practice back in their era. Notice Levine got the close results with the woman’s serum 83/21, as LW with rabbit anti-rhesus sera 85/15.
Landsteiner and Wiener managed to relate the effects Levine and Stetson observed on the ill woman at Bellevue to their newly named Rh Factor test discovered in the lab. Next, Wiener published an additional paper that identified a new, lethal blood factor responsible for transfusion reactions, and he had a blood test that identified this incompatibility.(5) This was a big discovery, except Levine had already made it, but didn’t name it.
Meanwhile, Levine was called in again to consult on a baby that died of Erythroblastosis fetalis by a Doctor Burnham.(1) According to Zimmerman, Burnham said Levine didn’t even know what Erythroblastosis fatalis was. He explained the disease to Levine. So, Levine used serum from the parents and dead baby for testing and concluded the mother had anti-Rh antibodies in her serum, and that caused the fetal death. In his paper Levine wrote “fetal antigens had provoked an antibody response in the mother, the same antigens had destroyed transfused blood from the father, and the same antigens destroyed the fetus’s blood before birth.” (6) The father and the infant had the same blood factor and this factor caused a reaction in the mother. Levine summed it up as a genetic trait. He offered a new model for Rh disease. Another major discovery that sadly, few believed! His discovery was the first to suggest that the baby wasn’t totally walled off from the mother’s blood, and that immune reactions weren’t limited to transfusion reactions as previously thought. Remember this comment in the Introduction? > In error, the paternal antigen (Rh+) was named the Rhesus factor.
Levine’s paper is the first one to connect the paternal antigen (Rh positive blood) to the Rh factor. He made the best correlation he could with the facts he had at the time. Years later, it was discovered the mother’s antibodies were produced against a different antigen from the one Landsteiner and Wiener discovered.
Together, but in separate labs, Levine and Wiener did outstanding work on Rh discoveries. Erythoblastosis fetalis was renamed Rh Hemolytic disease. Once a lab test was developed and connected to a disease affecting mother-baby, the medical world set out to end the suffering. They based their vaccine work on Levine’s discovery because his writing related Rh incompatibility to an immune response affecting mothers and their babies. To Levine, the other lab model of Landsteiner and Wiener didn’t fit with real people.
Levine went to work for Ortho Pharmaceuticals to create a vaccine for Rh disease, and Wiener stayed on with the New York City Medical Examiner’s office.
To recap: After 1940, for at least twenty years, people thought the two men discovered the same antigen at work and the term Rh became firmly entrenched in the mind and literature as the reason for maternal-fetal incompatibility. This turned out not to be the case. This turned out not to be the case! Please reread until this important fact and let it sink in. Here’s what happened next.
At first Wiener’s rabbit anti-rhesus serum and Levine’s serums from Rh negative women were both used by blood banks and labs. The problem with Levine’s formula was over time the antibodies would drop in the woman’s blood, plus they didn’t store well. The problem with Wiener’s test was it used animals and blood banks didn’t keep animals at hand. Blood banks needed quick access to large amounts of the anti-sera for blood transfusion testing. With World War II looming, they needed a way to produce a lot of test serum.
Wiener and Diamond discovered at the same time they could make large batches of test serum by injecting Rh positive red blood cells into Rh negative men without ill effects on the volunteers. (Men don’t have babies) During WW II, Wiener prepared anti-Rh serum for the armed forces by injecting a small amount of Rh positive red blood cells into people who were already sensitized by pregnancy or transfusion (men and women). Over time he found the best source of anti-Rh serum came from male Rh negative volunteers immunized by Rh positive red blood cells. (7)
This was Wiener’s procedure: Give at least two injections, small doses of .5cc (10 drops of blood) at 4 months apart to get specific high-titer anti-Rh antibodies. As long as the male test volunteers weren’t given an incompatible blood transfusion, the test gave them no ill effects. (8) By 1950 blood banks had abandoned the rabbit anti-rhesus sera reagents. Wiener’s second procedure on male volunteers is followed to this day.
As the blood tests got better, other scientists found other variants of the Rh Factor. Wiener claimed the new types of Rh factor came from one gene. He labeled them r’’ R1, R2, r, r’ etc. Scientists found his nomenclature confusing.(9) An alternative theory about Rh genes came from Sir Ronald Fisher. He claimed there were three genes on one chromosome responsible for Rh. He called them C c, D d, E e, after Landsteiner’s alphabet method. Wiener and another Rh expert Robert Race discovered separately but at the same time there were two forms of Rh antibody instead of one. The conclusion: More tests had revealed the antigens and antibodies recognized in the Levine-Stetson model and Landsteiner-Wiener model were not the same. (10) Owen
To summarize, the words Rh factor and anti-Rh introduced by Landsteiner & Wiener have persisted despite the heteroantibody they discovered being renamed anti-LW, and the human alloantibody being renamed anti-D. (7)Schwartz
Science writers today still mix up the terms. Scientists need clear models to follow, so they made the name change to make the differences clear. They never meant to confuse the lay public; they were making it clear for other researchers to follow. The rhesus-rabbit-human reaction seen by Landsteiner and Wiener in the lab were given the symbol LW (Landsteiner-Wiener). Their discovery is now considered genetically independent of Rh. This has been true for thirty years.
Because the term Rh had become public knowledge and associated with a mother-baby disease and the focus of a world-wide vaccine, the testing used by Levine and Stetson kept the Rh wording. Remember, Levine’s testing technique had nothing to do with the Rhesus monkey. Levine wrote the paper detailing his opinion, and his peers agreed with him based on all the testing.(11) Please note Levine’s title of his article in Science. (A D-like antigen in rhesus red blood cells and in Rh positive and Rh negative blood cells). <Myth busting!
See why there is so much confusion on the topic? The blood test with rabbit anti-rhesus factor isn’t being used, and rhesus monkey blood has nothing to do with mother-baby immune reactions. (12) Levine
For those of you who want to dive into the medical grist of this outcome see Catron and Rouger 1995, but this is academic work, not in laymen’s terms.(13)You’ve been warned!
Because Wiener named the Rh Factor, he felt threatened. His insecurity related in Zimmerman’s book is heart-breaking but funny. Both Levine and Wiener wanted recognition for their hard work. The question came down to the basic question.
Who discovered the Rh Factor?
Their blood feud grew so loud it embarrassed peers and friends. The feud prevented either one from ever winning a Nobel Prize, but both were nominated and both deserved it. According to Zimmerman, “While the Nobel Foundation doesn’t reveal losers names, five men from three nations have unasked, volunteered, the information that both Levine and Wiener were jointly nominated, but their quarrel compromised their chances of winning.” (1)Page 329
Next article: HDN Hemolytic Disease of the Newborn.
1. Zimmerman, D. 1973. Rh: The Intimate History of a Disease and Its Conquest. New York, Macmillan Publishing Co., Inc.
2. Levine, P, Stetson, R.E. 1939. An unusual case of intra-group agglutination. J. Am. med. Assoc. 113:126-127.
3. Landsteiner, K and A.S. Wiener. 1940. An agglutinable factor in human blood recognized by immune sera for Rhesus blood. Proc. Soc. Exp. Biol. Med. NY 48 223-224.
4. Wiener, A. 1938. The agglutinogens M and N in anthropoid apes. Journal of Immunology, 34, 11.
5. Wiener, A.S., Peters, H.R. 1940. Hemolytic reactions following transfusions of blood of the homologous group, with 3 cases in which the same agglutinogen was responsible. Ann. Intern. Med., 13:2306.
6. Levine, P., Katzin, E. and Burnham, L. 1940. Proc. Soc. Exp. Biol. Med. 45: 346-348.
7. Schwartz, H. P., and Dorner, F. 2003. Historical Review of Karl Landsteiner and his Major Contributions to Haematology. British Journal of Haematology 121: 556-565.
8. Wiener, A.S. 1945. Conglutination test for Rh sensitization. J. Lab. Clin. Med. 30:622
9. Wiener, A. 1969. History of Rh-Hr blood group system. New York State Journal of Medicine, Nov. 2915-2935.
10. Owen, R. 2000. Karl Landsteiner and the First Human Marker Locus. Genetics 155:995-998.
11. Levine, P. et al., 1961. Science. 133:323-333. A D-like antigen in rhesus red blood cells and in Rh positive and Rh negative blood cells.
12. Levine, P., Celano MJ, Wallace, J., Sanger, R. 1963. A human-like D like antibody, Nature. 198:596-597.
13. Cartron, J.P., and P. Rouger, Editors, 1995. Blood Cell Biochemistry, Vol. 6: Molecular Basis of Human Blood Group antigens. Plenum Press, New York.