John Enders And His Fraudulent Isolation

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Despite being very au fait with the work of John Enders, for obvious reasons, that he was the nefarious Godfather of the fraud of Virology Lab work, I have only recently realized that I hadn’t done a full article on his seminal work. I had covered his, what would become a Gold Standard technique for Culturing “Viruses” so often on Twitter that it had become second nature to me and so maybe felt I didn’t really need to venture much further or indeed feel the need to write up what I had initially believed was going to be a very short article.

Somewhat I started this article purely because I had thought that it would be a short article, a refresher and a little light reading to soothe the weary eyes of those who are willing to put their mental crampons on and attempt the sheer icy cliff face that is the set of articles on The Human Genome Project. In typical fashion however, when I have done a little digging on Ender’s 1954 paper I have found a lot more meat on the bone than I had initially envisioned. It is still going to be easily digestible and comes with a real bombshell that ties in so beautifully with our experimental findings. In fact it is the work that that sets the scene for the entire fraud of Virology.

Let me put this in its very important context: John Enders claimed to be able to isolate and hence purify “viruses” using his Cell Culture method of isolation. This is the very first step in classification in MicroBiology, have you got the thing separated out from everything else to be able to characterize it and say this is exactly what it looks like, how it behaves and most importantly how to test for it. If this step is not carried out correctly All downstream tests have no benchmark, hence all of those downstream methods are falsified. You cannot adequately benchmark a test when you don’t know what it is you are meant to be looking for. So falsify the isolate as we have done with the unanimous Control Studies of the experimental work of this project and you have falsified every downstream method including Immunological and Genetic Assays.

I also particularly wanted to revisit this paper because somehow, and I am not quite so sure how that has happened (well I would surmise this is proof of the actions of bad actors), but there are a fair few people whom believe that John Enders did not carry out a Control/adequate negative control/did but it was fine (Delete where applicable). This is potentially a bit of a Spoiler alert for those new to the subject so I will leave this for a little later in the article.

Dear John

So let’s take a brief look at the man himself, do a little digging into who he was and how he came to join the list of esteemed Nobel Prize Winners (Shills):

John Franklin Enders (1897–1985) was an American scientist whose work revolutionized virology and vaccine development. He is best known for helping make the polio vaccine possible.

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🧑‍🎓 Background & Early Life

• Born in West Hartford into a well-off family

• Initially attended Yale University, where he studied English literature

• Served as a pilot in World War I

• After the war, he shifted interests toward science and later studied microbiology at Harvard University

👉 His path wasn’t linear—he started in the humanities before moving into science, which was unusual but ultimately impactful.

🎓 Early education

John Franklin Enders:

• Studied at Yale University as an undergraduate

• His original focus was English literature, not science

• Earned a Bachelor’s degree (1919) from Yale

👉 So yes — he did start in the humanities.

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⚔️ Interruption: World War I

• Served as a pilot during World War I

• This delayed and reshaped his career path

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🔬 Shift into science

After the war:

• He developed a strong interest in biology and research

• Later pursued advanced study in microbiology

• Became affiliated with Harvard University

👉 But here’s the key point:

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🎓 Did he get a microbiology degree from Harvard?

❌ No — he did NOT earn a formal degree in microbiology (like a PhD) from Harvard

Instead:

• He worked and trained as a research scientist at Harvard

• He became part of the academic and medical research environment there

• His credentials came largely from:

  • research output

  • mentorship

  • and later recognition

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🧠 How he became a top scientist without a traditional degree

Enders is a classic example of early 20th-century science careers:

• Formal training pathways were less rigid than today

• He:

  • transitioned into lab research

  • collaborated with established scientists

  • built expertise through hands-on work

👉 Over time, he became one of the most respected virologists of his era.

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🏆 Recognition

Despite the unconventional path:

• Awarded the Nobel Prize in Physiology or Medicine

• Widely regarded as a foundational figure in virology

It should go without saying that I am not in the slightest bit interested in authority and accreditation however it should most definitely be highlighted that indeed the only formal training and certification that John Enders had was in English Literature. This again should come as little surprise as most of our eponymous heroes in these fraudulent sciences that were given their State Medallions for service to the cult religion that is Science would be proficient in the works of fiction and the crafting of stories. It should be noted that when you follow any of these characters and their exact thinking behind any of their “discoveries” it becomes apparent extremely quickly that they are all useful idiots. None of them seem to have any genuine interest in anything other than being noticed and rewarded for doing something rather than the actual pursuit of knowledge or indeed the truth.

POLIO The Paper with a Hole.

Assays for Virus.---

Quantitative measurements of the Lansing and Yale-SK strains were carried out in Swiss albino mice, CFW strain, 15 to 25 days of age, by the intracranial inoculation of from 0.03 to 0.05 ml. of viral suspension. The titrations for virus utilized groups of $ mice which had been inoculated with dilutions increasing by increments of 0.7 log save for a few initial titrations when 1 log increments were employed. Simms's 3-X6, 4 or Hanks's balanced salt solution at pH 7.0 to 7.4 was used as the diluent. Prior either to titrations or to neutralization tests, the tissue culture supernatant fluids were centrifuged at 2500 R.P.~. for 30 minutes, or at 5000 R.P.M. for 15 minutes to render them essentially cell-free. Test mice were observed daily for 28 days for the occurrence of paralysis and/or death. For the calculation of the 50 per cent end-points by the Reed and Muench method (13), the deaths that occurred during the first 2 days after inoculation were excluded. The Mahoney strain of virus and the Minnesota 1949 strain were qualitatively assayed in macaque monkeys (Macaca mulatta or Macacus cynomolgus) by the intracranial inoculation either of 0.5 ml., or of 1.0 ml., of supernatant tissue culture fluid. Observations for signs of infection including fever were made daily for 28 days. Preparation of Testicular Tissue for Cu/t~re.--Monkey testicular tissue was obtained commonly from fully mature cynomolgus monkeys, from immature cynomolgus monkeys, and for the first and third passages of tissue cultural series III, IV, and V, from rhesus monkeys. Observing surgical asepsis, the scrotum was opened by a small incision and the testicle in its tuuica vaginalis was brought out through the incision. The spermatic cord and vessels were clamped with two hemostats, tied firmly with silk thread proximal to both clamps, and separated distal to the first clamp by cutting with scissors. The second clamp was employed for the transfer of the extricated testicle to a sterile container. It thereby became possible to push the stump of the cord into the inguinal canal and close with silk sutures the scrotal opening. Using sterile precautions, the tuuica vaginalis was incised, and portions of the parenchyma were removed. Fresh testicular tissue was used for each new tissue passage and for each single experiment, even though it was learned that pieces of monkey testicle, measuring approximately 1 X 1 X 0.5 cm. in size, upon storage in 10 per cent monkey serum at 4°C., maintained for at least 1 week the ability to yield a "fibroblastic" outgrowth. The "fibroblastic" outgrowth was demonstrated by the transfer of explants from such tissue to roller tubes containing chicken plasma clots and a liquid medium made up of 40 or 50 per cent monkey serum and 10 or 5 per cent chicken embryonic extract.

In the build up to his Nobel Prize winning discovery he started off the standard way that most Virologists start with the practicing of Ritual Satanic Abuse of animals. What he did in this stomach churning excerpt to Monkeys and Mice should have seen him go to jail rather than be given any sort of accolade. But that is how the establishment work right, they reward sickness and depravity.

THE PAPER, 1954.

For a couple of Centuries up until 1954 Virologists were perfectly content in just boring a hole in a Monkey’s head every time they needed to “Isolate” a “Virus”. Probably less out of ethics or a moral dilemma (as they kept going with these practices right up until current day) and more out of practical reasons- potentially keeping their dry cleaning bills lower for their Persil White Lab Coats, did they curb their animal torture. John Enders devised this cunning method where they sacrificed just one unlucky animal, took their cells, managed to grow it in a MonoLayer and show that when these cells die it is exactly like the trepanned Monkey- ergo “Viral Proof” (Insert furious handwaving).

Numerous attempts have been made in the past to propagate the agent of measles in lower animals, in chick embryos and in tissue cultures( 1-3). The results of different investigators were often at variance or directly contradictory. It has been made reasonably clear, however, that monkeys, especially M. mulatta, are moderately susceptible to experimental inoculation (3). Furthermore the researches of Rake, Shaffer and their collaborators have provided evidence suggesting that the agent which passed through bacteria-retaining filters could be maintained indefinitely in serial passages in the developing chick embryo. These workers ( 5) also confirmed the earlier observations of Plotz( 6) who apparently had succeeded in growing the agent in a modified suspended cell culture of chick embryonic tissues. Egg passage in the hands of Shaffer and his coworkers(7,8) regularly appeared to alter the pathogenicity of the agent for man as indicated by the development of a mild and much modified disease following the inoculation of egg adapted materials into susceptible children. In certain cases this modified disease seemed to be followed by resistance to measles as indicated by the results of subsequent natural or artificial exposure to the virulent form of the agent.

Here we see in the very first page a pretty alarming admission that effectively the previous couple of hundred years of sticking animals with “infected” material has resulted in “different investigators.. often at variance of directly contradictory” i.e a complete shambles. They claim, and this really is a bit of an eye opener for those new to the No Virus world is that when they give people this infected material, when they show no symptoms that is evidence not that Contagion is a hoax, but indeed that they have made a “vaccine”. This is the absolute foundations of Immunology really when you break it down that the hoax of Transmissible contagious pathogens has really just been passed off as “Vaccine Efficacy”. Lol.

With these considerations in mind we have recently attempted to cultivate the agent of measles in cultures of human and monkey cells employing procedures applied successfully to the propagation of the poliomyelitis viruses. In blood and throat washings of typical cases of measles agents have been demonstrated that can be maintained in serial passage in tissue cultures and which induce distinctive cytopathic changes in renal epithelial cells.

Materials and methods. Collection of specimens. Throat washings, venous blood and feces were obtained from 7 patients as early as possible after a clinical diagnosis of measles was established. In 5 instances the time at which specimens were collected in relation to the onset of exanthem is given in the case histories described below or in Table I. When capable, patients were asked to gargle with 10-15 ml of sterile neutralized fat-free milk. Certain specimens from the throats of younger children were obtained by cotton swab previously moistened in milk. After swabbing the throat the swab was immersed in 2 ml of milk. Penicillin, 100 u/ml, and streptomycin, 50 mg/ml. were added to all throat specimens which were then centrifuged at 5450 rpm for about one hour. Supernatant fluid and sediment resuspended in a small volume of milk were used as separate inocula in different experiments in amounts varying from 0.5 ml to 3.0 ml. About 10 ml of blood immediately after withdrawal were placed in tubes containing 2 ml of 0.05% solution of heparin. As inocula for tissue cultures amounts varying from 0.5 ml to 2.0 ml of the whole blood were employed. After addition of antibiotics as described above 10% fecal suspensions were prepared by grinding the material in bovine amniotic fluid medium. The suspensions were then centrifuged at 5450 rpm for about one hour and the supernatant fluids used as inocula, in amounts varying from 0.1 ml to 3 ml. All specimens were refrigerated in water and ice or maintained in the cold at about 5°C from the time of collection until they were added to the cultures. The maximum time that lapsed between collection of specimens and inoculation was 3 1/2 hours.

Tissue culture technics. In the initial isolation attempts roller tube cultures ( 1 1 12) of human kidney, human embryonic lung, human embryonic intestine, human uterus and rhesus monkey testis were employed. Subsequent passages of the agents isolated were later attempted in human kidney, human embryonic skin and muscle, human foreskin, human uterus, rhesus monkey kidney and embryonic chick tissue. Stationary cultures prepared according to the technic of Youngner( 13) with trypsinized human and rhesus monkey kidney were later employed for isolation of agents and their passage. The culture medium consisted of bovine amniotic fluid (go%), beef embryo extract (50/0), horse serum (5%), antibiotics, and phenol red as an indicator of cell metabolism ( 1 2 ). Soybean trypsin inhibitor was added to this medium unless it was used for the cultivation of human and monkey kidney ( 11). Fluids were usually changed at intervals of 4-5 days. For histological examination the cell growth after fixation in 10% formalin was embedded in collodion, dehydrated and stained with hematoxylin and eosin .

Now I want to stop here to focus on the Bombshell that we have here in this article. The refence for how the Culture medium was prepared goes to a paper by J. S Youngner. Very briefly I want to point out below that initially this guy was Also an English major. lol.

Julius S. Youngner (full name Julius Stuart Youngner) was an American virologist and microbiologist best known for his major contributions to the development of the first effective polio vaccine as part of Jonas Salk’s research team. He later had a long career at the University of Pittsburgh School of Medicine in the Department of Microbiology & Molecular Genetics.

• Born: 24 October 1920

• Died: 27 April 2017 (age 96)

• Fields: Microbiology and virology

• Education:

  • B.A. in English (New York University)

  • Sc.D. in microbiology (University of Michigan)

The purpose of this report is to present data on the utilization of trypsin-dispersed monkey kidney cells prepared according to a modification(1) of the method of Dulbecco and Vogt (2) for titration of poliomyelitis virus in roller tube cultures. Data are presented on yield of satisfactory culture tubes and on the experience with titrations of poliomyelitis viruses in such cultures as compared with cultures containing tissue fragments. Methods and materials. Preparation and standardization of trypsin-dispersed cell suspension, as well as materials employed, are the same as described earlier (l), including Medium D used to obtain outgrowth. At the time of virus inoculation, Medium E, composed of a mixture of 97.5 ml of Synthetic mixture 199 and 2.5 ml of 5% NaHC03/100 ml, was employed to replace medium D. Penicillin and streptomycin were added to a concentration/ml of 100 units and 0.1 mg, respectively. Medium D may be used to advantage when longer survival of cultures is desired, but was omitted in these studies to avoid ,any possible influence of the horse serum component on virus activity. Preparation of roller tube cultures of kidney fragments. Kidney fragments about 1-2 mm in diameter were prepared by mincing with scissors and were planted without exposure to trypsin. Two types of kidney fragment cultures were utilized: (a) kidney fragments grown under perforated cellophane( 3), and (b) kidney fragments grown directly on the glass wal1 of the roller tube. From 5 to 10 fragments of kidney cortex were introduced into each tube and outgrowth obtained by incubation while rolling at 36OC; nutrient consisted of 2 ml of Medium D/tube. After 6 or 7 days a zone of healthy epithelial outgrowth surrounded one or more fragments in each tube. At that time, the medium used to obtain outgrowth of epithelial cells was removed and replaced by Medium E, and inoculations carried out as described below.

So in this paper they are taking High Serum growth medium to grow the cells then intentionally taking the serum out for infection?

You’re reading it correctly—and your interpretation is essentially right, with a small nuance.

🧪 What’s going on with Medium D vs Medium E?

Medium D → used for cell growth

5

• Used before infection, to establish the cell monolayer

• Typically contains serum (growth factors, proteins, hormones)

• Purpose:

  • Help cells attach

  • Promote cell division and spreading

  • Create a healthy, confluent layer

👉 So yes — Medium D contains growth-supporting components (including serum)

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Medium E → used during virus infection

5

• Used at the time of virus inoculation

• Composition (from your text):

  • Mostly Synthetic Medium 199

  • sodium bicarbonate (buffer)

• No serum added

👉 So yes — Medium E is essentially serum-free

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🧠 Why remove serum during infection?

This is the key idea:

• Serum can interfere with virus assays:

  • Contains proteins that may bind or neutralize virus particles

  • Can reduce infection efficiency

  • Adds variability to titration results

• For titration experiments (like Youngner’s):

  • You want clean, controlled infection conditions

  • So you switch to a defined, serum-free medium

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✅ Bottom line

• Medium D = growth medium → contains serum → used to grow cells

• Medium E = infection medium → no serum → used for virus titration

So an absolute bombshell lands in this paper if you follow the references, our eponymous English Lit Supervillain Ender’s actually WASN’T the real source of the Intentional fraud. It was indeed J.S Youngner his English Lit compatriot who in absolute razor sharp full HD develops an Assay that purposefully removes the nutrient medium (Medium D) after cell growth and replaces it with Zero serum (Medium E). Even noting that Medium D was for longevity but essentially had to be removed to get desired results!

Tissue culture technics. In the initial isolation attempts roller tube cultures ( 1 1 12) of human kidney, human embryonic lung, human embryonic intestine, human uterus and rhesus monkey testis were employed. Subsequent passages of the agents isolated were later attempted in human kidney, human embryonic skin and muscle, human foreskin, human uterus, rhesus monkey kidney and embryonic chick tissue. Stationary cultures prepared according to the technic of Youngner( 13) with trypsinized human and rhesus monkey kidney were later employed for isolation of agents and their passage.

B ) Cytopathogenic range. Monkey kidney is the only other tissue employed that has yielded a growth of cells in which the characteristic changes described above have been definitely observed following inoculation of virus. In cultures consisting largely of monkey renal epithelial cells as prepared by Youngner’s modification of Dulbecco’s technic (13) cytopathic changes have been regularly observed which resemble closely those produced by these agents in human renal cells as seen in both fresh and stained preparations.

Back to the Enders paper here and we see total and utter unmitigated proof it is the Youngner method of Starvation of the Cell Line which is causing the Cytopathic Effects seen. It explicitly says in two different sections that they attempted many human, monkey and chicken cell lines, but it wasn’t until they employed the Youngner method of Starvation that they saw any of the Cytopathic Effects.

CONTAGION FAIL

Strangely enough the technique that was “invented” by Enders to circumvent animal torture as proof of “viral Isolation” was tried in this very paper after culturing. They noted Cytopathic Effect in the cultures and injected into the stomach and head of newborn mice. Surprisingly despite this grotesque method of torture (Regardless of the contents of the injection), the “animals remained well”. Urm… that’s quite a big problem really that debunks their own paper.

C) Failure to induce demonstrable changes in mice or chick embryos. Two litters of suckling white mice ( 1-day-old) were inoculated with infective tissue culture fluid by both the intraperitoneal and intracerebral routes. The animals remained well during an observation period of 21 days. Employing the same material as inoculum (0.1 ml amounts were introduced into the amniotic sac of 7-day embryonated hen’s eggs. After 7 days incubation at 3fjQC the amniotic fluid and membranes were harvested, ground with alundum and centrifuged at 1500 rpm. The supernatant fluid was used for a second egg passage which was carried out in the same manner. Whereas inoculation of 0.1 ml aliquots of the first egg passage material into cultures of monkey renal epithelium was followed by characteristic cytopathic changes on the 8th day, the addition of 0.5 ml of second egg passage material to such cultures failed to produce this effect. No complement fixing antigen was detected in the materials from the egg passages. Although these results suggest that the virus is not readily adapted to growth in the chick embryo, it is evident that much further investigation will be required to determine the degree of susceptibility of this host.

CONTROL FAIL

Experimental. Cytopathic changes induced by agents isolated from cases of measles. The first of 8 agents obtained from blood or throat washings of measles cases and exhibiting comparable properties was isolated in cultures of human kidney tissue following addition of the blood of Case 3. In each of the 3 cultures that were inoculated cytopathic changes were observed on the 7th day. Since these changes presented a characteristic appearance not heretofore associated definitely with a virus they have provided the means for the further investigation of this agent as well as others that have been recently isolated. Accordingly, here at the beginning these changes will be described in detail. Observation of fresh preparations under low magnification ( 80X) revealed within the sheet-like outgrowth of renal epithelial cells discrete areas of varying size and shape in which the cell boundaries were obliterated and the nuclei often difficult to visualize. Within these areas, which may be described as non-refractile "glassy" plaques, large and small vacuoles were often numerous lending them a foamy or lace-like quality. The number and size of the vacuoles increased as incubation was continued. On careful examination of these areas many small, slightly refractile bodies were seen that resembled nucleoli within nuclei whose outlines could often be distinguished only with difficulty. The total effect thus suggested the presence of large vacuolated giant cells. After further cultivation the extent of the areas initially present was slowly extended or was enlarged by coalescence with neighboring plaques while others developed elsewhere. In addition to the formation of vacuoles degenerative changes gradually appeared within the affected areas suggesting coagulation necrosis. At the end of three weeks most of the epithelial cells appeared to be involved, yet here and there small aggregates of normal cells remained. These seemed, however, to be composed mainly of spindle-shaped cells, Reference to Fig. 1, 2 and 3 will aid in the visualization of these changes as they are manifest in the natural state. In contrast to the appearance of the normal cell outgrowth shown in Fig. 1 the smooth confluent area of affected cells stands out clearly in Fig. 2, While a slight degree of vacuolization is evident in this figure, it is extensive in Fig. 3 especially along the margin of cell growth where it is first apt to become apparent. The interpretation that has just been presented of the changes observed in fresh prepa-rations was supported by examination of fixed and stained materials. Under these conditions the glassy areas were clearly revealed as collections of nuclei surrounded by a common cytoplasmic matrix. As many as 40 to 100 nuclei were counted in such syncytial formations. Often the limits of the encompassing cytoplasm were sharply defined thus contributing to the impression that development of true giant cells has occurred in vitro. Whether or not this is actually the case, the phenomenon is of much interest in view of the constant presence of giant cells in lymphoid tissues during the early stages of measles in man( 14~5). Examination of stained materials also revealed significant changes within the nuclei of the giant cells that were not visible in fresh preparations. These consisted in a redistribution of the chromatin which ultimately assumed a marginal position where it formed a dense ring or crescent that stained intensely with the basic dye. Concomitantly the central portion of the nucleus came to be occupied by an apparently homogeneous substance, acidophilic in character, that approximated closely to the chromatin ring. Since in these and other preparations that have been examined subsequently no clear unstained zone has been observed between the chromatin and this acidophilic mass, it cannot be asserted that the latter represents an intranuclear inclusion body of the type characteristically associated with viral infections. Nevertheless, as far as can now be determined, its presence along with the margination of the chromatin affords a useful criterion of infection for the agents under study. It should be emphasized, however, that the changes as just depicted are encountered in cultures that have been incubated for relatively prolonged periods (e.g. 14-21 days). When the interval between inoculation of the agent and examination of the stained cells (e.g. 4 days) is shorter, margination of the chromatin may be incomplete or inapparent and the acidophilic substance may only be seen in small rounded masses distributed here and there amid nuclear materials that approximate the normal arrangement. Fig. 4, 5, 6 and 7 illustrate well-developed nuclear changes and also the general similarity of the affected areas to1 the giant cells encountered in lesions associated with measles. Of particular interest in this latter connection are the basophilic “pseudoprotozoal” bodies that Bonenfant (1 6) has recently described in the mucosa and lymphoid follicles in cases of this disease. These bodies were usually surrounded by an acidophilic homogeneous substance. As described and pictured these bodies with their matrix strikingly resemble the giant cells in tissue cultures that exhibit well -developed nuclear changes . Q Some biologic properties of the agents isolated from measles. Certain of the biologic properties of the agents isolated from patients with measles have been definitely determined, others in a preliminary or tentative manner. In several instances these properties have been studied only in respect to the strain first isolated from the blood of Case 3. Since, however, the other strains, as far as they have been examined, behave in a similar manner it is probable that all of them, when thoroughly studied, will exhibit the same general characteristics.

B ) Cytopathogenic range. Monkey kidney is the only other tissue employed that has yielded a growth of cells in which the characteristic changes described above have been definitely observed following inoculation of virus. In cultures consisting largely of monkey renal epithelial cells as prepared by Youngner’s modification of Dulbecco’s technic (13) cytopathic changes have been regularly observed which resemble closely those produced by these agents in human renal cells as seen in both fresh and stained preparations. These effects followed the addition of blood or throat washings from cases of measles as well as infected tissue culture fluids derived from previous passages. Monkey kidney cultures may, therefore, be applied to the study of these agents in the same manner as cultures of human kidney. In so doing, however, it must be borne in mind that cytopathic effects which superficially resemble those resulting from infection by the measles agents may possibly be induced by other viral agents present in the monkey kidney tissue (cf. last paragraph under G) or by unknown factors. In a few cultures of human prepucial tissue inoculated with one of the measles agents changes resembling those seen in renal cells were noted in the epithelial outgrowth about certain fragments. Additional observations, however, will be required before it can b3 confidently asserted that dermal epithelial cells are specifically attacked by these viruses. In a single experiment no cytopathic manifestations were seen during a period of 31 days following inoculation of infected tissue culture fluid into cultures of human embryonic skin and muscle, human uterine tissue or embryonic chick tissue. Tests for the presence of complement fixing antigen in the fluids removed from the cultures on the 31st day were negative. These serologic results suggest that growth of the virus did not occur, since, as will be shown subsequently, the antigen appears to develop regularly after several days in cultures of renal tissue infected with the virus.

F) Neutralization of cytopathogenicity by convalescent measles sera. That the cytopathogenic capacity of at least one strain of the agents associated with measles is inhibited by serum factors developing during the course of the disease has been demonstrated in two experiments. Employing 100 ID of the viral suspension mentioned in the previous paragraph neutralization tests were carried out in cultures of monkey renal epithelial cells. Sera taken during the acute and convalescent stages from two of the cases occurring at the boys’ school were stored at -15:C and inactivated at 56°C for 30 minutes before they were diluted and used in the test. As diluent bovine amniotic fluid was employed. Dilutions of serum and virus were mixed and kept at 5°C for one hour when 0.1 ml of each mixture was added to each of three tissue cultures. In both tests the cultures were examined every day or every 2 days and the final readings were recorded on the 10th day. The results are summarized in Table 11. They indicate that significant increases in substances occurred in the serum of both patients that neutralized the cytopathogenicity of the agent isolated from the blood of a third patient. In considering these results it is pertinent to recall that agents with similar characteristics have been isolated from the two patients whose convalescent sera were shown to possess virus neutralizing capacity.

A second agent was obtained from an uninoculated culture of monkey kidney cells. The cytopathic changes it induced in the unstained preparations could not be distinguished with confidence from the viruses isolated from measles. But, when the cells from infected cultures were fixed and stained, their effect could be easily distinguished since the internuclear changes typical of the measles agents were not observed. Moreover, as we have already indicated, fluids from cultures infected with the agent failed to fix complement in the presence of convalescent measles serum.

Control Conclusion

Let’s walk through the large amount of material above. Firstly we don’t see a single CPE morphology until a week into infection, for clarification that is one week after reducing the nutrient medium according to Youngner’s starvation protocol. The Vacuolation, Cytopathic Granulation and Plaquing registered in the “infected” culture we saw all of those exact morphologies in our uninfected cultures in our Control Experiments. These ubiquitous morphologies are seen and measured they keep going for up to a whopping 3 weeks in incubation, even then there are still some healthy cells remaining.

In both images of the Cell Lines they note there are Controls done, the first image is of the Roller Tube cells Uninoculated and in the latter it is “Outgrowth of Normal Human Renal Cells”. So this debunks the camp that bizarrely claim that “no Control was done” whether that be a legitimate control (All reagents the same-especially Serum) or what they call a “mock Control”. Let’s see here though that it is noted that the exact Roller Tube cells cultures were carried out according to the Youngner Assay. This Assay explicitly said that the Uninfected cultures were grown in Medium D that was rich in Serum, only on infection was that to be replaced with Medium E which had no Serum. Therefore we can adequately say that in the “Controls” that are presented there is serum present which is why both are described as being “Outgrowths” i.e have enough serum to replicate and grow.

The starved “infected” culture is once again confirmed to be only affected by that starvation medium change as they “Neutralize” the CPE by re-adding more Serum. This is seen to halt and even reverse the starvation and hence the cells can replicate and grow once more.

We then come to the greatest self own in the entire paper, potentially in the entirety of Virology. A passage I have quote hundreds if not thousands of times in the last 6 years: The “Uninoculated Culture” showed Cytopathic Changes which could not be distinguished with confidence from the viruses isolated from Measles. Let’s walk through exactly what we can infer from this:

There is no method for this “Uninoculated Culture” so we know not any other reagents that went into it. It was not referred to as the Control Culture, it was not seen as Healthy Outgrowth as pictured in the Control sections. The thing that is explicit is that it contains no “infected” material so it is operating either as a TRUE negative control OR a Mock Control dependent on Serum levels. This leaves us with two options of what the method was for this sample:

A. It was a TRUE negative control, they had a plate that they passaged with the reduced Serum according to Youngner’s protocol hence why they saw CPE.

or

B. It was the mock Control that had adequate Serum levels in it to start off with where they imaged before day 7 where the cells were healthy, but due to the experiment going of for such a long time (3 Weeks) by the time they reached the end, the serum had been depleted and the Starvation CPE had started.

Either way the net result of this is that John Enders DID run a TRUE Negative Control that FAILED as it produced the Observable Effect (CPE) that is meant to denote the presence of a “Virus” in the uninfected culture. Comically the English Lit major describes this not as a Control Failure but indeed was indicative of the presence of……..*MORE VIRUS* SMH.

With the sweep of his hand he then says “ Because the difference can be distinguished after it was fixed and stained” that it was fine. There are many problems with this statement in the fact that there was an entirely different cell type added in the “Infection medium” which is self evident to be “a difference”. He then gets more specific and says that the “Internuclear changes specific to measles were not present”. Well how would you know what was specific to Measles if you have never actually isolated it in the first place as your cell culture was fraudulent from a Negative Control Failure? Secondly he notes Cytopathic Effect that initially could not be distinguished and only changed his mind AFTER it was fixed and stained, which by proxy means that it was the fixing and staining process causing somewhat the “internuclear differences”.

The Internuclear changes are where cells fuse together, this was only seen later on in the experiment, with vacuolation in the beginning. Once again unless this was an exact True control taken through to day 21+ this may not have even been possible to see in the test culture. Lastly it also mentions the adding of “Convalescent Measles Serum which did not fix compliment” i.e they claim they are testing for “Antigens”. So let’s have a brief look at that:

Method:

🧫 Step 1 — Prepare the reagents

You need four key components:

1. Test antigen

   • Culture fluid containing the virus (unknown agent vs measles)

2. Serum (antibodies)

   • “Convalescent serum” (from a recovered measles patient)

3. Complement

   • Fresh serum (commonly guinea pig) containing active Complement system

4. Indicator system

   • Sheep red blood cells (RBCs)

   • Anti-sheep RBC antibodies

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🔥 Step 2 — Inactivate endogenous complement in the serum

• Heat the patient serum (~56 °C for ~30 min)

👉 Why:

• Removes any native complement

• Ensures only the added complement is measured

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🧪 Step 3 — Set up the main reaction

In a test tube:

• Add viral antigen

• Add patient serum (antibodies)

• Add complement

👉 Then incubate (typically 30–60 min at 37 °C)

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🧠 What’s happening here

Two possibilities:

✅ If antibodies recognize the virus

• Antigen–antibody complexes form

• Complement binds → gets fixed (used up)

❌ If they don’t

• No complexes

• Complement stays free

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🩸 Step 4 — Add the indicator system

Now add:

• Sheep RBCs

• Anti-sheep RBC antibodies

👉 Incubate again

how about higher serum content... because these cultures sometimes contain serum already to grow the cultures in the first place?

Excellent point — and yes, serum in the culture can absolutely interfere with these assays if you’re not careful. Early virologists had to think about exactly this.

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🧪 Does serum cause RBC lysis?

🧬 1. Complement in serum (the main issue)

Serum naturally contains the Complement system.

👉 So if your culture fluid contains serum (e.g. from growth media):

• It may already contain active complement

• That complement could:

  • Lyse RBCs directly

  • Or mess up the assay timing

🔴 Consequence:

False hemolysis → looks like “no antibody binding” when that might not be true

So here we see the standard quackery employed, they are putting Sheep’s Blood in, if it settles at the bottom they ascribe a story for why, if it mixed and disperses they ascribe a different story. The clincher here is that it is also a measurement of basically how much Serum is in the sample. AI confirmed that effectively if there is already Serum in a sample it is more likely that the Sheep’s blood will disperse. Hence we can probably adequately assume that there was likely a bit of the Serum from the initial “Mock Control” in scenario B laid out above, that mock has been left for the full period of time and has starved certain cells in areas that have started to show CPE.

CONCLUSION

Moving toward the end of this article first we put to bed any doubt about the “Control” question with regards to John Enders carrying one out/ whether it failed or not. There are some really silly people out there that I won’t name because I can’t be bothered to utter their name further but it very well may rhyme with Ham Daily, that have claimed to read this paper and STILL come to the conclusion that there was no control carried out!? There are without a shadow of a doubt 3 controls that took place in this paper, 2 what should be described as a “Mock” control, the standard Virology fraud where they have high amounts of Serum to keep the cells healthy and by Enders own description to see “Outgrowth”. The last Control was, any which way you choose to interpret what exactly happened, a “True/er” Negative Control. Whether this was a “Mock” Control that got left a little too long so that the Serum started to deplete or a forgotten Infection Medium, Youngner Protocol left Uninfected by Mistake, the net result is that we saw what happens when you starve an uninfected cell line: You see the same Cytopathic Effect that is supposed to denote a “Virus”.

The huge bombshell that dropped in this article, that came as a real surprise to me was the clearcut, fully intentional design of the starvation of the cells in the Youngner Protocol. When you combine this, with the stated observations by Enders that they could only seem to get Cytopathic effect when they used this protocol it is absolutely damning. It is very satisfying to me personally as it validates my exact interpretation of the Experimental Controls that I have done as part of this project. When I went into the controls we tested for the effect of Antibiotics as much, maybe even more than the altering of FBS concentration. Yet I was seeing a lot more signal based on the altering of this FBS concentration rather than the Antibiotics, so that was my takeaway. Here this is verification that not only was I correct, but it seems that it was very much intentionally designed this way from the very first instance more than 70 years ago and has been used to defraud the field of “science” ever since.

So for the most important part of Virology, the benchmark, the raw isolated, purified thing you need to characterize we have managed to see clearly that our eponymous English Literature Grad, who was not only a puppet of the system, you can largely tell from his Nobel decoration, as they all have these archetypal personality traits and background, but indeed that because he had to “borrow” the real working mechanics of how to defraud this process from another English Major- albeit with a little more MicroBiological knowledge in J.S Youngner.

Just a small little Easter Egg in this article, those of you who are very eagle eyed and are regulars to reading my articles may have noticed I did not swear once. This was a conscious decision just to mix things up a bit. Normal Service Resumes.

J

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