Was Influenza C Ever Purified and Isolated?

While looking into the origins of the influenza “virus,” it became clear to me that the particles claimed to be the swine flu, influenza A, and influenza B had never been properly purified nor isolated directly from the fluids of a sick host. These “viruses” had never been proven pathogenic in a natural way either, unless you believe injecting ground up animal organs mixed with various chemicals into other animals in different ways and then killing them, grinding up their organs, and repeating the process over and over again is “natural.” So it shouldn’t come as any surprise to learn that the same lack of evidence and grotesque animal experimentation applies to influenza C as well.

Trying to dig up information on the discovery of influenza C was more challenging than I initially thought it would be. Unlike influenza A and B which are regularly discussed and it was relatively easy to track down who was credited with their discoveries, influenza C seems to be the untalked about black sheep, the red-headed step child, the crazy uncle, etc. Uncovering not only who is credited with its discovery but also when it was discovered was fraught with uncertainty.

Was it discovered in 1951?

“A third form, C, was discovered in 1951. Influenza A is responsible for major epidemics and pandemics and influenza A is the only form that is found both in animals and in humans. The outbreaks of B tend to be smaller and less severe than those of A. Influenza C does not cause epidemics and causes only mild infections.”

https://www.google.com/url?sa=t&source=web&rct=j&url=https://law.yale.edu/sites/default/files/area/center/privatelaw/document/kitler_influenza_and_the_work_of_the_who.pdf&ved=2ahUKEwiOtIPk25jxAhV0FVkFHdMaBDA4ChAWMAh6BAgGEAI&usg=AOvVaw2xWQIAJZOLEKljzs1ZQN7E

Or was it 1947?

“Influenza C virus was first isolated in the forties (1947), with sporadic cases reported (10).”

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC104849/

Interestingly, upon researching, I came across two different names associated with the supposed discovery of influenza C. It was either first “isolated” by R.M. Taylor in 1947 under the name of strain 1233 or it was later “isolated” by “Mr. Influenza” himself Thomas Francis Jr. with the JJ strain in 1951. Oddly enough, even though Francis Jr. is given credit for the “isolation” of both A and B, it seems he must not have wanted the accolades associated with the discovery of influenza C as this is never mentioned in any of his biographies that I have seen. I only found out about his connection to the C strain through a citation in a paper by Taylor in 1951. Let’s see what we can uncover about this “discovery” by first going through some highlights from R.M. Taylor’s paper.

With Taylor’s work, you will find similar “isolation” methods to those employed for the previous versions of influenza. It involved taking throat washings from a patient who had mild symptoms which cleared the next day and mixing that with the blood of a horse as well as phosphate buffered saline. 200 ml of penicillin was added to this mixture before inoculating it into chick embryos. It was incubated for four days and then passaged (i.e. transferred) to another chick embryo. While no “virus” was visualized, the presence of a “virus” was recognized in the second egg passage by the agglutination of chicken erythrocytes (i.e. the clumping together of red blood cells). As no attempts to purify and isolate the particles assumed to be a “virus” were made, the subsequent antibody results (which are also unproven theoretical constructs) are meaningless.

Even if we were to (wrongly) assume that Taylor and others properly purified and isolated influenza C, he admits that electron microscope images by some researchers showed particles that had filaments while other researchers found particles with no filaments. This means that there were different particles being claimed as the same “virus” by different researchers which obviously shows the samples were not purified. The researchers each picked out particles which fit the preconceived “viral” representation that they wanted to find.

Attempts to infect animals with the “virus” failed miserably on numerous occasions. They tried experimenting on ferrets, hamsters, monkeys, mice, and rabbits and could not cause any symptoms of disease nor any histopathological findings of disease. All Taylor and Co. could find in some cases were rises in theoretical antibodies that the researchers had assumed were specific to a “virus” they had never seen. The question of human infectivity also remained unresolved. One researchers attempted to infect 6 women with a nasal spray of the JJ strain. Four of the six women had an antibody response yet none of the women became ill with any symptoms whatsoever. Taylor admitted that it was an assumption that an antibody response was related to infection and that they could not prove this to be the case, thus nullifying the meaningless theoretical antibody measurements.

While the above information should be enough to conclude that Taylor (and by extension Francis Jr.) never purified and isolated the influenza C “virus,” Taylor admitted that the position of both the 1233 and JJ strain as a disease-producing agent among human populations was uncertain. In other words, there was no proof they had a “virus.” He claimed that the presence of HI antibodies in a significant number of people in the United States implied that the “virus” had been widely dessiminated and that infection with it occurred frequently, yet the “virus” had only been linked to illness in a handful of cases. Taylor believed that the frequency of high HI antibody levels and the failure to connect the “virus” with symptomatic cases implied that many asymptomatic “silent” infections occurred. As can be seen, even when the evidence is insurmountable that no “virus” was ever recovered, virologists will always fall on the scapegoat of the unproven theoretical antibodies to “prove” the existence of unproven theoretical “viruses” in order to claim healthy people are sick. Highlights are presented below:

A Further Note on 1233 (“Influenza C”) Virus*

“From human throat washings there was isolated in 1947 a virus which appeared to be antigenically unique and different in some other respects from inflnenza A and B viruses and their variants. The strain was designated as Strain 1233, the serial number of the throat washing from which it was secured. Further details concerning this strain were to be published later, but up to the present this has not been done, primarily because from available data it was not possible to assess the public health importance of this virus. Moreover, active laboratory work on respiratory viruses including the 1233 strain was discontinued at the close of 1949.

Although the frequent presence of rather high hemagglutination inhibition (I-II) titers to 1233 in human sera suggested extensive seeding of this virus in the northeastern part of the United States, it could be associated, by means of HI titration of acute and convalescent phase sera, with only three of 185 otherwise undiagnosed upper respiratory infections accompanied by manifest symptoms. This led to the conclusion that Strain 1233 “may prove to be of little public health importance,” since from the health aspect only infections which give rise to disease or health impairment are of particular interest.

Francis and his associates isolated later a virus (J J) which proved to be “essentially identical” with the 1233 strain, and by retrospective serological diagnosis they identified it with a previous upper respiratory episode in a home for young children. They verified the distinctive qualities of this virus and proposed that it be classified as “influenza C.”

It is the purpose of this communication to present in greater detail the author’s experiences with this virus as well as to refer to the important contributions since made by others in classifying the status of this virus.

Methods.

The methods used in the isolation and study of this virus were analogous to those commonly employed in the investigation of the A and B type influenza viruses. A 2 percent normal, heat-inactivated horse serum in a phosphate buffered saline solution was used in securing the throat washings (TW) and in suspending material for subsequent passage in chick embryos and other animals. Routinely, 200 units of penicillin were added per ml. TW prior to inoculating chick embryos. Each embryo received an inoculum of 0.2 ml. The Salk modification of the Hirst HI test with four hemagglutinating units of antigen was employed. Antigens of the 1233 strain were prepared from the amniotic fluid of infected chick embryos. Following addition of the chicken erythrocyte suspension the tubes were immediately placed in a cold room at about 4°C. This is important in working with the 1233 virus. The complement fixation (CF) tests were performed in the usual manner with two units of complement. While infected allantoic fluid was not used as a source of antigen for the HI test with the 1233 virus, it was usually found to be satisfactory for CF antigen.

Isolation of Virus. The virus was obtained from a TW taken on March 19, 1947 from an adult male (JMP) living in New York City. On the day the TW was taken the patient complained of a mild headache, backache and coryza, suggesting the onset of a cold or influenza attack. The temperature was not taken. On the following day, the symptoms had subsided and he continued his usual office duties with no further discomfort. The TW was inoculated into the amniotic sack of chick embryos within a few hours after being taken. After four days incubation of the embryos at 36°C., the amniotic fluid was harvested. The presence of a virus was recognized in the second egg passage by the agglutination of chicken erythrocytes. The same virus was isolated on two later occasions from a portion of the TW which had been stored in a lusteroid tube in a C02 ice box at approximately –78°C. The subsequent behavior of the virus in chick embryos will be described later.

Unfortunately, means for securing a blood sample were not at hand when the TW was taken. It was the intention to obtain a sample on the following day, but when the patient reported the disappearance of symptoms it was concluded that he did not have an influenza attack and an acute phase blood sample was not taken.”

“About the time the 1233 virus was isolated upper respiratory illness was quite prevalent in New York City and vicinity, and eleven other virus strains, all classified as influenza A, were isolated. Yet, no other infection by the 1233 virus was recognized during this period, either by immune response or virus isolation.”

Size and Morphological Characteristics. Filtration through graded gradocol membranes indicated that the 1233 virus is approximately the same size as influenza A virus. This was confirmed by Francis, et al. 2.
Electron microscope pictures made in these laboratories by Dr. J. C. Bugher verified the size of the virus and showed it to be of spheroid shape. No filamentous forms were observed. However, on some electron microscope photographs of the 1233 strain taken by Dr. J. B. Murphy, which he was kind enough to send to us, there appeared some filaments not unlike those observed in preparations of PR8 and other influenza strains. Francis, et al. also reported finding numerous filaments in infected chick amniotic fluid. Like other influenza viruses, the 1233 strain may be satisfactorily preserved in the frozen state at low temperatures and may be frozen and dessicated without material loss in infectivity titer.

Behavior in the Embryonated Chick Egg. The primary isolation of the virus was made by inoculation of TW into the amniotic sac of 9-day-old embryos, as no older embryos were available at that time. In subsequent passages of the virus, and in re-isolation from the TW, embryos of from 10 to 13 days of age were used. Within these limits the exact age of the embryo seemed to have little effect upon the growth of the virus in the amniotic sac. However, 11-day-old embryos were preferred for maintaining the virus in passage in the amniotic sac for two reasons: at this age the embryo is easier to inoculate by the technique employed, and a larger amount of amniotic fluid can usually be harvested than from either older or younger embryos. Dilutions of the amniotic fluid or washings from the amniotic sac served as the virus-passage material. During the earlier passages the eggs were incubated (36°C) for 3 to 4 days before harvesting. Later it was found that if dilutions of the amniotic fluid ranging from 10^-2 to 10^-5 were inoculated, two days incubation was sufficient to produce maximum increment of the virus. In fact it is preferable to harvest not later than 48 hours following inoculation, because the infection produces a contraction of the amniotic sac and a diminution of the fluid, which makes it difficult to obtain adequate material for passage and H (hemagglutination) titrations. Amniotic fluid from infected embryos may attain infectivity titers of 10 -9 and H titers of 1:16000. The virus produces Iittle or no mortality in embryos during the first five to six days of incubation following inoculation. Longer incubation periods were not tried.

During the first 15 passages by the amniotic route eight attempts were made to infect embryos by inoculation into the allantoic sac. On only one occasion did the harvested allantoic fluid agglutinate chicken erythrocytes, and then in a dilution not exceeding 1:16. Following 20 or more amniotic sac passages it was usually possible to demonstrate growth in the allantoic sac by means of hemagglutination of the harvested fluid. Continued serial passages in the allantoic sac gave inconsistent and unpredictable results: although hemagglutination in low titers was always obtained, the height of the titer was variable. In no instance did the H titer exceed 1:256, although the infectivity titer of the allantoic fluid commonly reached 10-7. Thus, while the virus may be serially propagated in the allantoic sac the allantoic fluid does not seem to be a dependable source of H antigen, although some batches were usable. It may be employed, however, for CF tests.”

“Hemagglutination peculiarities. The 1233 virus causes agglutination of chicken erythrocytes, and as with the other influenza viruses, this phenomenon may be used to recognize infection of embryonated eggs and to measure the HI antibody content of sera. But in comparison with the other influenza viruses, the behavior of the 1233 strain is erratic. During the early passages in embryos it was found difficult to properly standardize and preserve a hemagglutinating antigen. In some preparations the titer diminished rapidly irrespeetive of the temperature at which the preparation was kept. In others the titer remained constant over a considerable period of time. Later in the embryo passage series the hemagglutinating capacity of the virus became more stable, and by the 50th passage an antigen of high titer (1:2000 to 1:16000) and of good keeping quality could consistently be prepared from infected amniotic fluid. The virus elutes rapidly from the erythr0cytes at room temperature. For best results it is therefore essential to place the tube racks in an ice box at about 4°C immediately after adding the erythiocytic suspension. The erythrocyte suspension should preferably be chilled before’it is added to the virus mixture. The virus agglutinated guinea pig erythrocytes during the early embryo passages, although never in as high a dilution as with chicken erythrocytes. The capacity to agglutinate guinea pig erythrocytes diminished as the embryo passages increased. While the virus will grow in the allantoic sac and while infected allantoic fluid will produce agglutination of chicken erythrocytes, the titer reached in allantoic fluid is inconstant and much inferior to that reached in the fluid of embryos infected by inoculation into the amniotic sac.

It was subsequently noted by Hirst that agglutination of fowl erythrocytes by this virus is depressed by the presence of relatively small amounts of certain electrolytes, particularly calcium salts. This phenomenon may have accounted in part for the difficulties experienced in obtaining eonsistant hemagglutination during the early egg passages.

Infectivity for Laboratory Animals. Intranasal inoculation of ferrets, hamsters and a monkey evoked a specific rise in HI and CF antibodies to the 1233 virus. No definite symptoms were manifested by these animals, nor were any gross lesions noted in the turbinates or lungs of ferrets and hamsters autopsied on the fourth day following inoculation. Nevertheless, the rise in HI and CF antibodies implies that infection occurred. Francis, et al. 2 report that ferrets inoculated with JJ virus had mild fever but showed no other clinical evidence of infection; their convalescent sera reacted equally well to JJ and 1233 viruses but not to A, A’ and B strains.

Due to a shortage of ferrets no efforts were made to continue passage of the 1233 virus in this animal. The 1233 virus was, however, carried through three serial passages in hamsters. Fourth passage animals failed to become infected. Three attempts to establish the virus in mice by blind lung-to-lung passage met with failure. No reproducible lung lesions occurred, nor was the virus recoverable from the lungs of the mice by inoculation into chick embryos. Mice are not susceptible to intracerebral inoculation of the virus. Inoculation of the cornea of a rabbit’s eye by scarification produced no manifest lesions.”

Human Injection and Symptomaology. The crucial questions concerning this virus from the aspect of human health are: how frequent are human infections and what symptoms or health impairment does infection
produce? Neither of these questions can be adequately or comprehensively answered at present, but some suggestive information is at hand.

If it be assumed that infection with this virus gives rise to a specific antibody response, and that the presence of humoral antibodies to this virus is indicative of a previous infection with it, then the antibody level of a population should be a measure of the frequency of past infections. While these assumptions cannot be proven to be correct, all experimental and observed evidence support their validity.

The following tabulation reflects the results of HI titration of sera from 204 adults residing principally in the northeastern part of the United States (Table VII).

It will be observed that only 26.5 percent gave a titer of less than  1:32 while 62.2 percent had a titer of 1:64 or above, and 42.1 percent had a titer equal to or exceeding 1:128. Unfortunately, it is not known what the titer should be in a normal, or previously unexposed, person. Francis, et al. found that the HI antibody level in young children (in an institution) preceeding an influenzal outbreak in which this virus (JJ strain) was involved was usually below 1:32. It seems not improbable, therefore, that unexposed persons possess little or no demonstrable HI antibody.

It will be recalled that the HI titer of the person from whom 1233 was originally obtained dropped from 1:256 to 1:64 within twelve months after the infection. It seems not improbable, therefore, that the persons (62.2 percent.) who had a titer of 1:64 or above had been infected previously with this virus. The data, although limited in scope, suggest that the majority of adults have had some experience with this virus; the high titers encountered in some of the sera suggest that the experience was of recent date. Francis, et al. reached the same conclusion after examining adult sera from the student population at the University of Michigan. It should not be inferred, however, that this virus is a recent arrival: a serum taken in 1942 gave a titer of 1:1024. Further evidence on human susceptibility to infection with this virus was recently contributed by Quilligan, et al. Six adult women volunteers received nasal spray inoculations of the JJ strain; four of the six subsequently showed a specific antibody response to the virus; the person who administered the spray developed clinical signs of influenza and also showed a specific antibody response to JJ.

When the 1233 strain was isolated it was hoped that it might account for many of the clinically manifest acute upper respiratory infections that were not attributable to the hitherto known influenza virus types. Our efforts to connect it with such infections were disappointing. Of 185 paired sera from patients with undiagnosed respiratory disease, only three pairs showed a rise in HI titer to this virus following the attack (Table VIII). It is of interest that two of the three sera showing an immune response to 1233 were among the three specimen pairs received from Venezuela. The remaining 182 paired sera came from the northeastern United States, and among these only one pair indicated that 1233 had been responsible for the infection. Practically all of the 185 sera came from adults, including the three that gave positive response. Hirst, who examined sera from 50 patients in New York City with undiagnosed respiratory infections, could connect none of the infections with this virus. Nor were Francis, et al. able to demonstrate any rise in HI titer to the JJ strain in 52 University of Michigan students with respiratory infections, although a majority of the sera did show a typical rise in titer to virus strains of the A-A’ group.

On the other hand, Francis, et al. found a significant rise in HI titer to JJ strain among 1- to 5 1/2 year-old children in a children’s home following an influenzal outbreak. However, A’ infection was also associated with the epidemic, and it is not clear from the data presented how many of the symptomatic infections were attributable to A’ and how many to JJ virus.

Thus, the only recorded symptomatic infections due to the virus in question are: the very mild infections in the two adults from whom the 1233 and JJ strains were obtained; the three cases listed in Table VIII; the clinical infection in the person who administered the nasal spray to Quilligan’s six volunteers (none of the four in this group who showed an antibody rise had manifest symptoms); and, subject to interpretation, the children’s home episode. It should also be mentioned that some of the adult attendants in this home also showed a rise in antibody to the JJ strain following the epidemic (Francis, et al)).

Discussion and Gonclusions.

The prototype 1233 strain and the seemingly identical JJ strain appear to represent a species or a type of virus not hitherto described. This is evidenced by its lack of any detectable antigenic relationship to the previously known A-A’ and B influenza viruses or to mumps and
Newcastle disease viruses. The type specificity of the 1233 strain is further demonstrated by the unique character of its hemagglutination receptors
and inhibitors in relation to other members of MNI group. It also possesses certain peculiarities with regard to its behavior in embryonated eggs and its requirements for the effective agglutination of chicken erythrocytes.

It is similar, however, to the previously known types of influenza virus in its ability to infect (production of a serological immune response) when inoculated intranasally into ferrets, hamsters, monkeys and human beings (JJ strain). It also resembles previously known types in regard to the lung lesions and other cytological reactions it produces in embryonated eggs when it is inoculated into the amniotic sac.

Its position as a disease-producing agent among human populations
is still uncertain. While the presence of HI antibodies in a significant number of people in the United States implies that the virus has been widely dessiminated and that infection with it occurs frequently, the virus has been definitely linked with manifest illness in only a few instances. The symptoms in the reported infections have been mild but still compatible with the clinical diagnosis of influenza.

The frequency of high HI antibody levels and the failure to connect the virus with any but a few symptomatic infections imply that many asymptomatic silent infections occur. These facts suggest an agent of high infectivity but low virulence or pathogenicity. This characteristic in turn may explain why the virus has not been encountered previously and more frequently in throat washings, the search for virus in throat washings usually being made only in the presence of symptoms of upper respiratory disease. It is conceivable, however, that in unconditioned persons, such as young children, or in populations not previously exposed to it, the virus may give rise to epidemics of symptomatic disease. This epidemic among children reported by Francis, et al. and the serological identification of two clinical infections in Venezuela furnish grounds for being on the alert for upper respiratory disease produced by this virus in young children and in populations that may not have been previously exposed to the virus.

It is of interest and perhaps of significance that both the 1233 and the JJ strains were isolated at a time when infections with A and A’ influenza viruses were prevalent. Likewise there was an A’ influenza epidemic in the children’s home during the period when rises in HI titers to the JJ strain occurred 2. From this circumstantial evidence it might be surmised that the factors which influence the spread of A and A’ influenza also contribute to infections with the 1233-JJ strain.

As regards nomenclature we agree with Hirst and with Francis, et aI. that this virus deserves a specific designation. On the basis of receptors and inhibitors Hirst favors regarding it as a new member of the MNI group. However, in human infections attention must be given to the
nature and epidemiology of the disease complex. In this respect it resembles the influenza viruses more closely than it does any of the other viruses in the MNI group. Consequently we are inclined to approve the proposal of Francis, et al. that it be named Influenza C.”

doi: 10.1007/BF01241168.

Now that we have some background on how Taylor and Francis Jr. both apparently discovered the same “virus” at different times through different methods which can only be related to each other through assumption-filled and flimsy serological tests, let’s look a bit at Francis Jr.’s 1950 account. Here you will see that Francis Jr. claimed to have isolated a “virus” by amniotic inoculation of chick embryos from fresh throat garglings of a single ill person. Being the expert virologist that he is, Francis Jr. dodged providing specific materials/methods used for the “isolation” of his JJ strain of influenza C. His evidence is based solely on antibody results. Even then, he found the same positive antibody results in those who were said to have influenza A as those who supposedly had influenza C. Perhaps this is why Taylor himself claimed that Francis Jr.’s work was “subject to interpretation.” Francis Jr. even admitted that Taylor concluded that influenza C “may prove of little public health import,” and Hirst noted no significant rises of antibody to it. Thus we can see that the 1233/JJ drama associated with influenza C is nothing but a dud:

Identification of Another Epidemic
Respiratory Disease

“A mild outbreak of influenza associated with A-prime strains of virus occurred in Ann Arbor, Michigan, in the spring of 1950, and several characteristic strains were isolated. On March 20, a virus unrelated to known strains of the influenza group was isolated by amniotic inoculation of chick embryos from fresh throat garglings of a person ill on March 16. The illness experience was similar clinically to a mild form of the influenza prevalent at the time and consisted of one day of headache and malaise, with short intervals of myalgia, but no fever was noted. Acute and convalescent blood specimens were obtained from the patient and showed rises in hemagglutination-inhibiting (HI) titer from 64 to 2,048, in complement-fixing titer from 0 to 64, and in neutralizing titer in eggs from 128 to 650. Similar tests against A, A-prime, and B strains of influenza virue showed no rises. This information clearly indicated the specificity of the virus (JJ) in relation to the illness.

In an effort to determine the relationship of this virus to the A-prime epidemic, tests were made with 52 pairs of sera that had been obtained in the acute and convalescent stages from patients among the students at the University of Michigan, ill with influenza during the epidemic phases of 1947 and 1950. None of them showed rises in titer to the JJ virus, although a majority did show typical rises in titer to strains of virus of the A-A-prime influenza group. However, it was noted that most of them did show moderately high HI titers to the JJ virus, suggesting that they had previously had experience with a virus of this type; a few of them had sufficiently high titers to suggest that their experience had been recent.

The possibility existed that the adults, because of the high titers, might not show a measurable response to infection with this virus. Attention was then directed to young children who would have less experience, and whose antibody responses would be more likely to reflect the occurrence of infection. Sera obtained from a group of children, aged 1-5 1/2 years, who had been studied from August, 1946, to May, 1947, were available. They had been vaccinated with the PR8 strain of Type A influenza in the fall of 1946, and blood samples were obtained at intervals from August to November, 1946, and again in the latter part of April and early May, 1947 (1). During March and April, 1947, a sharp outbreak of A-prime influenza occurred. The serum samples from August through November could be compared with those obtained in May, 1947, for antibody rises against strains of influenza virus isolated from tho epidemic. This was done and showed clear evidence of the prevalence of A-prime influenza in the Children’s Home. The same sera were tested for HI antibodies against the JJ virus and, surprisingly, a large number of them showed a pronounced rise in antibodies in the sera taken in April and May, indicating that there had been a wide exposure to this virus in the interval between November and May.

Because of their broader range of reaction, the extensive tests made in the present study with human sera are of still greater significance. Although 32 of the children from 1947 developed antibodies to both A-prime and JJ viruses, the remaining 36 showed specific reactions in which no cross relation between Types A, A-prime, or B influenza virus and JJ virus was detectable by HI, complement-fixation, or neutralization tests in eggs. Patients from the influenza B epidemic of 1945 (3) revealed no rise in antibody to the JJ virus. Moreover, subjects vaccinated against PR8, FAI1, or Lee strains exhibit no rise in antibodies to the JJ virus. The virus under consideration thus seems to be serologically and immunologically distinct from previously identified strains of influenza virus.

The possibility remained, however, that it might be related to some other well-established clinical disease. Pairs of acute and convalesceut sera were examined from cases of rubeola, rubella, infeetious mononucleosis, atypical pneumonia, and the common cold; with negativeresults. A group of sera from adults with unidentified respiratory disease, furnished by J. E. Salk, of the University of Pittsburgh, gave no positive results. Sera positive to JJ virus were negative to Newcastle disease virus. Rabbit antimumps serum, mouse sera against Melnick’s Texas, Ohio, and Connecticut strains of Coxsackie virus, and mouse anti-PVM serum were all negative to the JJ virus. The egg membranes or embryos exhibit no lesions representative of the pox groups of virus. LCL bodies have not been seen, but electron microscopy of amniotic fluid has disclosed numerous filaments. There is, therefore, in these results no evidence that this virus is related to the common infectious diseases mentioned above. Hirst suggests on the basis of the action of this virus upon cell receptors and egg-white inhibitor that it may be unrelated to known members of the group of hemagglutinating respiratory viruses (4).

Although Taylor concluded that the virus “may prove of little public health import,” and Hirst noted no significant rises of antibody to it, the present data indicate that it is causally related to a widespread respiratory disease that occurs in epidemic form. Neither Taylor nor Hirst refers to the high incidence of antibodies in the adult population, which to our minds strongly indicates that the population has been thoroughly seeded. On the other hand, in young children titers are generally extremely low and, when present, tend to be quite high, suggesting recent infection. Moreover, tests with sera from as far back as 1936 indicate that the virus has been circulating since that time at least. At present, the frequency with which this virus causes clinical disease is difficult to estimate, but the epidemic herein described not only involved children but also a significant number of adults among the limited number in the institution. The clinical features in adults are not yet well outlined, but fever, cough, and coryza in the children were the common signs. The study emphasizes a frequently neglected opportunity to clarify epidemiological problems by the study of young children.

The association of the epidemic disease with influenza, the basic clinical picture, and the wide distribution of antibodies in the human population, as well as the serological and immunological characteristics of the virus, readily invite consideration of the name “Influenza C.” Further studies, a number of which are under way, will determine the appropriateness of this suggestion.

doi: 10.1126/science.112.2913.495.

In Summary:

• Influenza C, a third form of the influenza “virus,” was discovered in 1951…or maybe it was “isolated” in 1947…who knows…?
• It was either isolated by R.M Taylor or Thomas Francis Jr. or both

• From human throat washings there was isolated in 1947 a “virus” which appeared to be antigenically unique and different in some other respects from inflnenza A and B “viruses” and their variants
• The strain was designated as Strain 1233
• Further details concerning this strain were to be published later, but up to 1951 had not been done, primarily because from available data it was not possible to assess the public health importance of this “virus”
• Moreover, active laboratory work on respiratory “viruses” including the 1233 strain was discontinued at the close of 1949
• 1233 could be associated with only three of 185 otherwise undiagnosed upper respiratory infections accompanied by manifest symptoms
• This led to the conclusion that Strain 1233 “may prove to be of little public health importance,” since from the health aspect only infections which give rise to disease or health impairment are of particular interest (i.e. it was not associated with disease…hence not a “virus”)
• Thomas Francis Jr. and his associates isolated later a “virus” (JJ) which proved to be “essentially identical” with the 1233 strain, and by retrospective serological diagnosis they identified it with a previous upper respiratory episode in a home for young children
• A 2 percent normal, heat-inactivated horse serum in a phosphate buffered saline solution was used in securing the throat washings (TW) and in suspending material for subsequent passage in chick embryos and other animals
• Routinely, 200 units of penicillin were added per ml. TW prior to inoculating chick embryos and each embryo received an inoculum of 0.2 ml
• On the day the TW was taken the patient complained of a mild headache, backache andcoryza, suggesting the onset of a cold or influenza attack
• The temperature was not taken and on the following day, the symptoms had subsided and he continued his usual office duties with no further discomfort
• The TW was inoculated into the amniotic sack of chick embryos within a few hours after being taken
• After four days incubation of the embryos at 36°C., the amniotic fluid was harvested
• The presence of a “virus” was recognized in the second egg passage by the agglutination of chicken erythrocytes
• In other words, they looked at an effect and assumed the presence of an invisible “virus” based on “the clumping together of cells or particles, especially bacteria or red blood cells, usually in the presence of a specific antibody or other substance” https://www.thefreedictionary.com/agglutination)
• It was the intention to obtain a sample on the following day, but when the patient reported the disappearance of symptoms it was concluded that he did not have an influenza attack and an acute phase blood sample was not taken
• About the time the 1233 “virus” was isolated upper respiratory illness was quite prevalent in New York City and vicinity, and eleven other “virus” strains, all classified as influenza A, were isolated and yet no other infection by the 1233 “virus” was recognized during this period, either by immune response or “virus” isolation
• Filtration through graded gradocol membranes indicated that the 1233 “virus” is approximately the same size as influenza A “virus”
• Electron microscope pictures made in these laboratories by Dr. J. C. Bugher verified the size of the “virus” and showed it to be of spheroid shape with no filamentous forms
• However, on some electron microscope photographs of the 1233 strain taken by Dr. J. B. Murphy, there appeared some filaments not unlike those observed in preparations of PR8 and other influenza strains
• Francis, et al. also reported finding numerous filaments in infected chick amniotic fluid
• In other words, the “virus” did not look the same when the images were taken by different people and sadly, we have to take the word of the researchers as none of the non-identical EM images were published in the study…for some reason…
• The primary isolation of the “virus” was made by inoculation of TW into the amniotic sac of 9-day-old embryos
• Dilutions of the amniotic fluid or washings from the amniotic sac served as the “virus-passage” material
• The “virus” produced Iittle or no mortality in embryos during the first five to six days of incubation following inoculation and longer incubation periods were not tried
• During the first 15 passages by the amniotic route eight attempts were made to infect embryos by inoculation into the allantoic sac
• On only one occasion did the harvested allantoic fluid agglutinate chicken erythrocytes, and then in a dilution not exceeding 1:16
• Following 20 or more amniotic sac passages it was usually possible to demonstrate growth in the allantoic sac by means of hemagglutination of the harvested fluid
• Continued serial passages in the allantoic sac gave inconsistent and unpredictable results
• The 1233 “virus” was said to cause agglutination of chicken erythrocytes, and as with the other influenza “viruses,” they determined that this phenomenon may be used to recognize infection of embryonated eggs and to measure the HI antibody content of sera
• But in comparison with the other influenza “viruses,” the behavior of the 1233 strain was erratic
• During the early passages in embryos it was found difficult to properly standardize and preserve a hemagglutinating antigen
• In some preparations the titer diminished rapidly irrespeetive of the temperature at which the preparation was kept
• The erythrocyte suspension was to be preferably chilled before’it is added to the “virus” mixture
• The capacity to agglutinate guinea pig erythrocytes diminished as the embryo passages increased
• While the “virus” was said to grow in the allantoic sac and while infected allantoic fluid was said to produce agglutination of chicken erythrocytes, the titer reached in allantoic fluid was inconstant and much inferior to that reached in the fluid of embryos infected by inoculation into the amniotic sac
• Intranasal inoculation of ferrets, hamsters and a monkey evoked a specific rise in HI and CF antibodies to the 1233 “virus”
• No definite symptoms were manifested by these animals, nor were any gross lesions noted in the turbinates or lungs of ferrets and hamsters autopsied on the fourth day following inoculation
• Nevertheless, they claimed that the rise in HI and CF antibodies implied that infection occurred
• In other words, there was no clinical nor histopathological signs of infection but the antibody results told them that this non-existent infection occurred
• The 1233 “virus” was carried through three serial passages in hamsters
• Fourth passage animals failed to become infected
• Three attempts to establish the “virus” in mice by blind lung-to-lung passage met with failure
• No reproducible lung lesions occurred, nor was the “virus” recoverable from the lungs of the mice by inoculation into chick embryos
• Mice were not susceptible to intracerebral inoculation of the “virus”
• Inoculation of the cornea of a rabbit’s eye by scarification produced no manifest lesions
• According to Taylor, the crucial questions concerning this “virus” from the aspect of human health are: how frequent are human infections and what symptoms or health impairment does infection produce?
• He admitted that neither of these questions could be adequately or comprehensively answered and only some suggestive information was at hand
• He stateed that if it is assumed that infection with this “virus” gives rise to a specific antibody response, and that the presence of humoral antibodies to this “virus” is indicative of a previous infection with it, then the antibody level of a population should be a measure of the frequency of past infections
• He then admits that these assumptions cannot be proven to be correct
• In other words, Taylor:
  1. Failed to make animals sick both clinically and histopathologically
  2. Failed to prove the frequency of human infections
  3. Failed to provide specific symptoms for his “virus”
  4. Failed to prove antibodies were specific and indicative of infection
• While Taylor tried to claim antibody levels as an indicator of infection, he admitted that it was not known what the titer should be in a normal, or previously unexposed, person
• Taylor claimed that because the HI titer of the person from whom 1233 was originally obtained dropped from 1:256 to 1:64 within twelve months after the infection, it was not improbable that the persons (62.2 percent.) who had a titer of 1:64 or above had been infected previously with this “virus” (i.e. he is guessing)
• He stated that it should not be inferred that this “virus” was a recent arrival as a serum taken in 1942 gave a titer of 1:1024
• Six adult women volunteers received nasal spray inoculations of the JJ strain; four of the six subsequently showed a specific antibody response to the “virus;” the person who administered the spray developed clinical signs of influenza and also showed a specific antibody response to JJ
• In other words, four women did not develop disease but had a “specific” antibody response while two women also did not develop disease and had no antibody response…i.e. none of the six developed disease and the antibody results are meaningless
• When the 1233 strain was “isolated” it was hoped that it might account for many of the clinically manifest acute upper respiratory infections that were not attributable to the “known” influenza “virus” types, yet Taylor admits that their efforts to connect it with such infections were disappointing
• Of 185 paired sera from patients with undiagnosed respiratory disease, only three pairs showed a rise in HI titer to the “virus” following the attack
• Hirst, who  examined sera from 50 patients in New York City with undiagnosed respiratory infections, could connect none of the infections with the “virus”
• Francis, et al. was unable to demonstrate any rise in HI titer 
  to the JJ strain in 52 University of Michigan students with respiratory infections
• However, Francis, et al. supposedly found a significant rise in HI titer to JJ strain among 1- to 5 1/2 year-old children in a children’s home following an influenzal outbreak
• However, A’ infection was also associated with the epidemic, and it was not clear from the data presented how many of the symptomatic infections were attributable to A’ and how many to JJ “virus”
• According to Taylor, the only recorded symptomatic infections due to the “virus” in question are: the very mild infections:
  1. In the two adults from whom the 1233 and JJ strains were obtained
  2. The three cases listed in Table VIII
  3. The clinical infection in the person who administered the nasal spray to Quilligan’s six volunteers (none of the four in this group who showed an antibody rise had manifest symptoms)
  4. Subject to interpretation, the children’s home episode
• As the children’s home is admittedly subject to interpretation, that makes a grand total of 6 symptomatic “infections” with numerous non-symtomatic infections based on unreliable and, contrary to their claims, entirely non-specific antibody results
• Taylor believed that the prototype 1233 strain and the seemingly identical JJ strain appeared to represent a species or a type of “virus” not described
• This was evidenced by its lack of any detectable antigenic relationship to the previously known A-A’ and B influenza “viruses” or to mumps and Newcastle disease “viruses”
• He claimed it was similar to the previously known types of influenza “virus” in its ability to infect (production of a serological immune response) when inoculated intranasally into ferrets, hamsters, monkeys and human beings (JJ strain)
• Its position as a disease-producing agent among human populations was uncertain
• Taylor claimed that while the presence of HI antibodies in a significant number of people in the United States implied that the “virus” had been widely dessiminated and that infection with it occured frequently, the “virus” had been definitely linked with manifest illness in only a few instances
• In other words, the only evidence to claim influenza C as a “virus” is theoretical antibody results
• The frequency of high HI antibody levels and the failure to connect the “virus” with any but a few symptomatic infections implied that many asymptomatic silent infections occur
• These facts suggested an agent of
  high infectivity but low virulence or pathogenicity
• Taylor believed that this characteristic may have explained why the “virus” had not been encountered previously and more frequently in throat washings, the search for “virus” in throat washings usually being made only in the presence of symptoms of upper respiratory disease
• From the circumstantial evidence, Taylor felt it might be surmised that the factors which influence the spread of A and A’ influenza also contribute to infections with the 1233-JJ strain (thus using previous fictitious strains to claim evidence of another fictitious strain)
• In human infections attention must be given to the nature and epidemiology of the disease complex
• In this respect, Taylor claimed it resembled the influenza “viruses” more closely than it did any of the other “viruses” in the MNI group and was inclined to approve the proposal of Francis, et al. that it be named Influenza C

• A “virus” unrelated to known strains of the influenza group was “isolated” by amniotic inoculation of chick embryos from fresh throat garglings of a person ill on March 16
• The illness experience was similar clinically to a mild form of the influenza prevalent at the time and consisted of one day of headache and malaise, with short intervals of myalgia, but no fever was noted
• Sera was taken from 52 students at the University of Michigan, ill with influenza during the epidemic phases of 1947 and 1950
• None of them showed rises in titer to the JJ “virus” although a majority did show typical rises in titer to strains of “virus” of the A-A-prime influenza group
• However, it was noted that most of them did show moderately high HI titers to the JJ “virus,” suggesting that they had previously had experience with a “virus” of this type
• The possibility existed that the adults, because of the high titers, might not show a measurable response to infection with this “virus”
• Attention was then directed to young children who would have less experience, and whose antibody responses would be more likely to reflect the occurrence of infection
• The same sera were tested for HI antibodies against the JJ “virus” and, surprisingly, a large number of them showed a pronounced rise in antibodies in the sera taken in April and May, indicating that there had been a wide exposure to this “virus” in the interval between November and May
• In other words, they are assuming non-specific antibody results prove the presence of the “virus” that they could not purify nor isolate from any of these cases
• Because of their broader range of reaction, Francis Jr. stated the extensive tests made in the present study with human sera were of still greater significance
• Even though the anibody results were seen for both A-Prime and JJ within the same samples, he claimed that the “virus” seemed to be serologically and immunologically distinct from previously identified strains of influenza “virus”
• The possibility remained that it might be related to some other well-established clinical disease
• Electron microscopy of amniotic fluid disclosed numerous filaments (even though no EM images were presented)
• Taylor concluded that the “virus” “may prove of little public health import,” and Hirst noted no significant rises of antibody to it
• The present data indicated that it was causally related to a widespread respiratory disease that occurs in epidemic form
• Neither Taylor nor Hirst referred to the high incidence of antibodies in the adult population, which to Francis Jr. strongly indicated that the population had been thoroughly seeded
• Tests with sera from as far back as 1936 indicated that the “virus” had been circulating since that time at least
• Notice that everything Francis Jr. claimed about this “virus” is based on non-specific antibody results and not purified/isolated “virus” particles
• Francis Jr. stated that the frequency with which the “virus” caused clinical disease was difficult to estimate
• The clinical features in adults were not yet well outlined, but fever, cough, and coryza in the children were the common signs
• The association of the epidemic disease with influenza, the basic clinical picture (which was unknown), and the wide distribution of (non-specific) antibodies in the human population, as well as the (non-specific) serological and immunological characteristics of the “virus,” was enough for Francis Jr. to submit for consideration the name “Influenza C”
• He then claimed further studies, a number of which were under way, would determine the appropriateness of this suggestion

If one is being intellectually honest, it is clear that influenza C is nothing more than an attempt to cover up for the fact that the tests designed to detect influenza A and B do not work every time, thus necessitating another strain to blame for the remaining cases of influenza-like illness that remain. However, these efforts were admittedly met with disappointment. The evidence presented for influenza C turned out to be just as faulty and fraudulent as that which was presented for A/B. Overall, what we have is two sets of teams looking at two different serially-passaged ground up tissue/chick embryo goo who performed assumption-based serological tests to determine if their strains were not only the cause of unknown outbreaks of influenza in the 40’s but also whether they were somehow related. Due to a lack of data supporting the public health importance of this “virus,” much of the data was not presented until the early 50’s when both men decided that they had the same “virus” and labelled it influenza C. Maybe it was due to the weak evidence of human illness? The lack of cases? The lack of symptoms? The unpredictable and erratic antibody results? Or maybe it was once again the lack of a properly purified/isolated “virus” taken directly from a sick parient and proven pathogenic in a natural way? Whatever the case may be, it seems no one wanted the credit for the discovery of the ugly blemish that is influenza C.