Did Thomas Weller Isolate the Varicella-Zoster “Virus” in 1954?

According to the CDC:

“In 1954, Thomas Weller used cell culture to isolate VZV from vesicular fluid of patients with varicella or zoster.”

https://www.cdc.gov/vaccines/pubs/pinkbook/varicella.html

Taking the CDC at their not-so-trustworthy word, the ‘virus” responsible for both the chickenpox and shingles was not isolated until 1954. This means that any paper said to be working with the “virus” up to that point, such as Thomas Rivers in 1923, could only assume that they were working with a “virus” as varicella-zoster was never isolated for over half of the 20th century. However, if we are being picky, the paper itself was actually published in 1953 and Thomas Weller was awarded the Nobel Prize for his disgusting tissue culture practices in 1954, yet seeing how the CDC also believes a 14-year-old Rudolph Steiner performed experiments and wrote a paper in 1875 proving chickenpox infectiousness, I should probably let this error slide.

In any case, here is a brief history on Weller’s work:

“In 1954, the Nobel Prize for Medicine was awarded to Drs John Enders, Thomas Weller, and Frederick Robbins for their watershed discovery that growth of poliomyelitis virus occurred in cultures of cells of extraneural origin, first reported in 1949. Their demonstration in 1949 that the Lansing type II strain of poliomyelitis could be grown in cultures of human embryonic tissue set into motion a race to develop a vaccine for the disease that had crippled countless thousands of individuals. The discovery and subsequent recognition were only the beginning of a prolific career for Thomas Huckle Weller, who made numerous contributions to the field of virology, including isolating the varicella-zoster virus (VZV) from cases of chickenpox and zoster, providing suggestive evidence that the same virus is responsible for both diseases; isolating the human cytomegalovirus (CMV) for the first time in tissue culture and suggesting the descriptive name now used for it; establishing Coxsackie viruses as the cause of epidemic pleurodynia: and first isolating rubella virus, the cause of German measles.”

https://pubmed.ncbi.nlm.nih.gov/12118846/

“Suggestive evidence.” That is a very firm conclusion drawn there. Keep in mind that this “suggestive evidence” stems from the same sick tissue culture practices of aborted human embryos and foreskin tissues mixed with numerous chemicals and sources of animal DNA that was also used for the “isolation” of Polio. It must also be kept in mind that what virologists mean by “isolation” is the exact opposite of the accepted meaning of the word “isolation.”

So did Weller really “isolate” VSV and provide “suggestive evidence” as stated in his brief biography and by the CDC? Below is his full paper from 1953 with highlights and a summary:

Serial Propagation in vitro of Agents Producing Inclusion Bodies Derived from Varicella and Herpes Zoster.

It is generally accepted that serial propagation in the laboratory of the agents of varicella and of herpes zoster has not heretofore been accomplished. Certain of the earlier published reports regarding their serial propagation have more recently been attributed to possible confusion with viruses pathogenic for lower animals. However, morphological evidence has suggested that a single passage of the agents of varicella and herpes zoster has been achieved. Thus Rivers ( 1,2 ) observed focal lesions with intranuclear inclusions in the monkey testicle following the local inoculation of varicella vesicle fluid. Goodpasture and Anderson (3) grafted human skin on the chorioallantoic membrane of the chick embryo. Although negative results were obtained following inoculation of varicella vesicle fluid, in one experiment histological examination of grafts following inoculation with fluid from herpes zoster lesions revealed intranuclear inclusions. Using essentially the same technic, Blank, Coriell and Scott(4) likewise in one instance demonstrated inclusion bodies after inoculation of zoster material.

In 1948 we began an investigation of the potentialities of suspended cell cultures of human tissues as a medium for the isolation of the causative agent of varicella. Eosinophilic intranuclear inclusions were demonstrated on examination of tissue fragments removed from the cultures at intervals after the introduction of varicella vesicle fluid( 5). Efforts to maintain the responsible agent on serial passage in this type of culture were unsuccessful. More recently roller tube cultures of human tissues have been applied to the same objective. The results so far obtained are here reported in a preliminary manner. As will be shown it has been possible to isolate and maintain in serial passage cytopathogenic agents apparently derived from specimens of varicella vesicle fluid as well as others apparently obtained from herpes zoster vesicle fluid.

Materials and methods. Vesicle fluid specimens were collected and stored as previously described ( 5). Roller tube cultures were prepared as in our studies on poliomyelitis (6) utilizing either human embryonic skin-muscle tissue or foreskin tissue obtained from boys between the ages of 3 months and 3 years. In the present experiments the nutrient fluid for the cultures consisted of bovine amniotic fluid (90%), beef embryo extract (5%), horse serum (SP), antibiotics, soybean trypsin inhibitor and phenol red as recently described (6,7). Changes of the nutrient fluid were made at 3- or 4-day intervals. In certain experiments for histologic examination tissues were grown on coverslips coated with chicken plasma that were placed in roller tubes; these preparations were fixed with Zenkers-acetic acid and stained with hematoxylin and eosin.

Experimental. 1 ) Studies with varicella
vesicle fluid. Vesicle fluids derived from 11 cases of varicella were inoculated individually, usually in the form of 0.1 ml aliquots of a suspension in milk, into groups of roller cultures of human embryonic skin-muscle tissue. The exact concentration of the fluid in milk cannot be stated. In every instance tissue growth was well established at the time of inoculation. As summarized in Table I, in cultures inoculated with fluids from six of these cases, focal cytopathogenic lesions of a
characteristic appearance developed which were readily seen on microscopical examination of the living cultures. These lesions usually were first observed from the 6th to 8th day after inoculation. They consisted of small collections of swollen, rounded refractile cells which contrasted sharply with the surrounding fibroblastic or epithelial outgrowth. Those foci of affected cells developing in the sheets of cells of normal appearance increased slowly in size. The cells in the center of such focal areas gradually degenerated over the course of several days, while slow peripheral extension of the lesion continued for days or weeks as contiguous cells became infected. Study of stained coverslip preparations from roller tube cultures inoculated with vesicle fluid material has shown that the changes in cellular morphology are characteristically associated with the presence of inclusion bodies. At the margin of a focus occasional cells may be observed that show no gross morphological changes, yet contain small granular intranuclear inclusions. Rounded and swollen cells almost invariabIy possess intranuclear eosinophilic inclusions. Occasionally the larger swollen cells are multinucleate, with each nucleus containing an inclusion; such cells resemble the multinucleate giant cells described by various workers in the lesions of varicella, herpes zoster and herpes simplex (8). Focal lesions developing in a pre-existing loose network of cells progress irregularly, but manifest similar changes. The morphological changes observed in the cultures will be reported in more detail in a future communication.

All six of the agents isolated following the inoculation of varicella fluids have been propagated serially in tissue culture as indicated by the successive development in subcultures of focal areas of cellular enlargement and degeneration. Two of the strains (McE. and Wel.) have now been maintained for 10 passages as summarized in Table 11. The inoculum employed to initiate the third and succeeding passages in these experiments consisted of 0.1 ml of a suspension of coarsely ground tissue removed from the preceding set of cultures. On one occasion, the infected tissue suspension was frozen in the COz box prior to use; otherwise, the inoculum was prepared on the day it was employed. Fig. 1 to 3 depict representative cytopathic changes observed during the serial propagation of the McE. and Wel. agents. In contrast to the results obtained with tissue suspensions, attempts to establish passages with inocula consisting of centrifuged fluids removed from the infected cultures have so far been unsuccessful.

Studies designed to elucidate the nature and etiological relationship of these cytopathogenic agents thus isolated to variceIIa are in progress. In one experiment, vesicle fluid material of known infectivity for cultures did not produce cytopathic changes when inoculated following heating at 60°C for 30 minutes. In none of more than 70 control cultures maintained during the passage experiments have focal lesions been observed; these tubes routinely received inocula consisting of a suspension of tissue derived from control cultures of the preceding passage. Tissue suspensions infective for cultures have been without obvious effect when inoculated into suckling mice or into the developing hens egg by various routes. Attempts at in vitro neutralization of the cytopathogenic effect with convalescent serum from cases of varicella have so far been unsuccessful. It is possible that the close association of the infective agents with cell constituents may be involved in this apparent lack of neutralizing effect. In certain preliminary experiments in which culture fluids harvested from tubes showing cytopathic changes have been employed as antigen in complement fixation tests on paired serum specimens obtained from cases of varicella, a rise in antibody titer has been observed. The specificity of this reaction is under investigation.

2) Studies with herpes zoster vesicle fluid.
Similar focal cytopathic changes have been observed in cultures inoculated with material obtained on the day of appearance of vesicles from an 80-year-old (Sto.) with thoracic herpes zostert and with fluid collected from a 30-year-old woman (Pie.) with involvement of the ophthalmic branch of the trigeminal nerve. The Sto. inoculum had been stored in the frozen state for 21 months prior to use, while the Pie. fluids were inoculated on the day of collection. Serial propagation in tissue culture of these two agents has been accomplished, as summarized in Table III, again with inocula consisting of ground tissue suspensions. (These manipulations have been performed in a separate room from that in which the varicella agents have been maintained.) The cytopathogenic effect observed in the cultures closely resembles that obtained following the introduction of varicella vesicle fluid. Examination of stained preparations from the second tissue culture passage of the Sto. strain has also revealed that the rounded swollen cells composing the focal lesions contain intranuclear inclusions (Fig. 4). Tissue suspensions prepared from cultures of the Sto. and Pie. agents, that were successfully employed to initiate culture passages, have produced no overt symptoms in newborn mice when inoculated by various routes.

Discussion. Inoculation of roller tube cultures of human tissues with materials derived from the eruptive lesions of varicella and of herpes zoster has revealed the presence of cytopathogenic agents capable of producing intranuclear inclusions. These, subsequently, have been maintained in serial passage. A peculiarity of their behavior in the culture system employed has been the apparent failure of infectious material to appear in the fluid phase. The local lesions appear to increase in size by infection of immediately adjacent cells. On prolonged cultivation, however, an increase in the number of focal lesions occurs. In one experiment in which infected cultures were maintained for 8 weeks approximately 90% of the proliferating tissue became involved. Even so, extension of the lesions was continuing at the end of this period. The significance of these observations is being investigated. It is at present clear, however, that in the utilization of tissue culture technics for the isolation of unknown agents, consideration must be given to passage of tissue suspensions, as well as to the passage of fluid inocula.

No evidence has been obtained that the agents isolated are not those responsible for varicella and herpes zoster. Yet no definitive statement can now be made regarding their nature, their etiological relationships, or the interesting question of the possible identity of the agents of herpes zoster and varicella. It is to be noted that the virus of herpes simplex when propagated in tissue cultures of the type here employed manifests an early focal tendency to induce lesions but then rapidly brings about a widespread infection of the cell population resulting in an appearance quite dissimilar to that here described (9). The failure of these agents to produce obvious infection of suckling mice, in view of the high degree of susceptibility of this animal to the virus of herpes simplex (10). also indicates that they are not to be identified with the latter. We have no indication that a Rickettsia-like agent of the type described by Sprunt and Hirst ( 11) was present in our cultures.

Summary. The inoculation of roller tube
tissue cultures of human tissues with vesicle fluid derived from patients with varicella has resulted in the isolation of six cytopathogenic agents. Two of the strains have been maintained for 10 tissue culture passages by employing tissue suspensions as the passage material. Histologically the lesions produced consist of focal accumulations of cells which become swollen, and then degenerate: characteristically, such cells contain intranuclear inclusion bodies. From the eruptive lesions of two cases of herpes zoster, cytopathogenic agents of a similar nature have been isolated and have been propagated serially in cultures of human tissue.

https://doi.org/10.3181/00379727-83-20354

In Summary:

• It is generally accepted that serial propagation in the laboratory of the agents of varicella and of herpes zoster has not been accomplished
• Certain of the earlier published reports regarding their serial propagation have been attributed to possible confusion with “viruses” pathogenic for lower animals
• Goodpasture and Anderson grafted human skin on the chorioallantoic membrane of the chick embryo
• Negative results were obtained following inoculation of varicella vesicle fluid yet histological examination of grafts following inoculation with fluid from herpes zoster lesions revealed intranuclear inclusions
• In 1948 Weller began an investigation of the potentialities of suspended cell cultures of human tissues as a medium for the isolation of the causative agent of varicella (thus discrediting any other so-called isolation attempts such as those by Rivers in 1923)
• Efforts to maintain the responsible agent on serial passage in this type of culture were unsuccessful
• Weller turned to roller tube cultures of human tissues to attempt the same objective
• Weller claimed that his results showed it was possible to isolate and maintain in serial passage cytopathogenic agents apparently derived from specimens of varicella vesicle fluid as well as others apparently obtained from herpes zoster vesicle fluid
• Roller tube cultures were prepared as in his studies on poliomyelitis utilizing either human embryonic skin-muscle tissue or foreskin tissue obtained from boys between the ages of 3 months and 3 years
• The nutrient fluid for the cultures consisted of:
  1. Bovine amniotic fluid (90%)
  2. Beef embryo extract (5%)
  3. Horse serum (SP)
  4. Antibiotics
  5. Soybean trypsin inhibitor
  6. Phenol red
• In certain experiments for histologic examination tissues were grown on coverslips coated with chicken plasma that were placed in roller tubes; these preparations were fixed with Zenkers-acetic acid and stained with hematoxylin and eosin
• Vesicle fluids derived from 11 cases of varicella were inoculated individually, usually in the form of 0.1 ml aliquots of a suspension in milk, into groups of roller cultures of human embryonic skin-muscle tissue
• The exact concentration of the fluid in milk cannot be stated
• Study of stained coverslip preparations from roller tube cultures inoculated with vesicle fluid material had shown that the changes in cellular morphology were characteristically associated with the presence of inclusion bodies
• Quick sidenote: Intranuclear inclusion bodies (INB) are aggregates of protein that are said to be frequently encountered in “viral” infections, where they are thought to be accumulations of “viral” particles, yet for RNA “viruses” replicating in the cytoplasm, this compartmentalization represents a paradox not consistent with the “viral” replication cycle https://pubmed.ncbi.nlm.nih.gov/1332364/
• The larger swollen cells seen wre multinucleate, with each nucleus containing an inclusion; such cells resembled the multinucleate giant cells described by various workers in the lesions of varicella, herpes zoster and herpes simplex
• All six of the agents “isolated” (I must have missed that part…) following the inoculation of varicella fluids were propagated serially in tissue culture as indicated by the successive development in subcultures of focal areas of cellular enlargement and degeneration
• The inoculum employed to initiate the third and succeeding passages in these experiments consisted of 0.1 ml of a suspension of coarsely ground tissue removed from the preceding set of cultures
• In contrast to the results obtained with tissue suspensions, attempts to establish passages with inocula consisting of centrifuged fluids removed from the infected cultures were unsuccessful
• Studies designed to elucidate the nature and etiological relationship of these cytopathogenic agents thus isolated to variceIIa are in progress
• In other words, Weller claimed he “isolated” particles of varicella but he had no idea whether they were in fact varicella
• In one experiment, vesicle fluid material of known infectivity for cultures did not produce cytopathic changes when inoculated following heating at 60°C for 30 minutes
• Controls consisted of tubes which routinely received inocula consisting of a suspension of tissue derived from control cultures of the preceding passage
• Tissue suspensions infective for cultures were without obvious effect when inoculated into suckling mice or into the developing hens egg by various routes
• Attempts at in vitro neutralization of the cytopathogenic effect with convalescent serum from cases of varicella were unsuccessful
• Weller claimed it was possible that the close association of the infective agents with cell constituents may be involved in this apparent lack of neutralizing effect (in other words, not purified/isolated)
• In some preliminary antibody tests, a rise in antibodies was observed, however, the specificity of this reaction was under investigation
• Similar (i.e. not identical) focal cytopathic changes were observed in cultures in two cases of herpes zoster
• Serial propagation in tissue culture of these two agents was said to be accomplished, again with inocula consisting of ground tissue suspensions
• These manipulations were performed in a separate room from that in which the varicella agents were maintained
• The cytopathogenic effect observed in the cultures closely resembled (again, not identical) that obtained following the introduction of varicella vesicle fluid
• Tissue suspensions prepared from cultures of the Sto. and Pie. agents, that were successfully employed to initiate culture passages, produced no overt symptoms in newborn mice when inoculated by various routes
• While Weller claims successful culture of cytopathic agents serially propagated in tissue culture, he admits a peculiarity of their behavior in the culture system employed was the apparent failure of infectious material to appear in the fluid phase
• He detailed changes in the tissue observed but states the significance of these observations were still being investigated
• Weller believed that in the utilization of tissue culture technics for the isolation of unknown agents, consideration must be given to passage of tissue suspensions (significance unknown), as well as to the passage of fluid inocula (material not infectious)
• No evidence had been obtained that the agents isolated are not those responsible for varicella and herpes zoster yet no definitive statement could be made regarding their nature, their etiological relationships, or the interesting question of the possible identity of the agents of herpes zoster and varicella
• The failure of these agents to produce obvious infection of suckling mice, in view of the high degree of susceptibility of this animal to the “virus” of herpes simplex, indicated to Weller that they are not to be identified with the latter
• In other words, Weller had no idea if he isolated the varicella-zoster “virus” and his “virus” did not produce varicella, herpes zoster, or herpes simplex in mice…meaning Weller failed…he should have led with that and saved us all some time…

In order to believe that Thomas Weller isolated the varicella-zoster “virus” and provided “suggestive evidence” that both shingles and chickenpox are caused by the same “virus,” we must do two things:

1. Change the definition of the word “isolation” to mean taking fluid from chickenpox/shingles cases and adding human foreskin tissue, beef amniotic fluid, beef embryo extract, horse serum, antibiotics, soybean trypsin inhibitor, and phenol red and then culture this concoction for days.
2. Completely disregard Weller’s own negative evidence of infectiousness as well as his statement that what he had created could not be claimed to be either varicella or herpes zoster.

“Suggestive evidence?” A more accurate description would be NO EVIDENCE.