Tissue Culture Techniques and Their Application to Original Isolation, Growth, and Assay of Poliomyelitis and Orphan Viruses

Melnick, Joseph L.

doi:10.1111/j.1749-6632.1955.tb42532.x

Cited by 136 publications

(34 citation statements)

References 29 publications

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“…For these viruses a lowering of the temperature from 37' increases the relative plating efficiency on cells not supplied with cystine. Substituting the growth medium of Melnick (1955) for medium D has a similar effect. These modifications may operate through altering the size of the intracellular pool of cystine or 'cystine-activity', the…”

supporting

confidence: 48%

“…screw-cap serum bottles with square cross-section. I n a few experiments the growth medium of Melnick (1955) was used in place of medium D. After this growth period the cultures were washed three times with cold phosphate-buffered saline (PBS) (Dulbecco & Vogt, 1954), the tube cultures each time with 2-0 ml., the plate cultures each time with 4-25 ml., and the bottle cultures each time with 10 ml. After virus inoculation, the cultures were maintained at 35-37'" on 0.90, 6.0 and 10 ml.…”

Section: Methodsmentioning

confidence: 99%

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Poliovirus Mutants with Altered Responses to Cystine

Dubes

Chapin

1958

Journal of General Microbiology

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SUMMARY: Three strains of poliovirus (Akron, antigenic type 1; Brooks, type 2 ; Mabie, type 3), which require exogenously supplied cystine (or a cystine-substitute) for optimal cytopathogenic action on monkey kidney tissue cultures, were serially passaged ten times in cystine-deficient monkey kidney tissue cultures in tubes. From various passage levels in this system were isolated three mutants of Akron, four of Brooks and two of Mabie. These nine mutants are designated the cystine-response mutants and are distinguishable from their ancestral viruses used to initiate the passage series by being relatively cystine-non-requiring. Two criteria suffice to distinguish the mutants of any strain from each other: (1) the degree of loss of requirement for cystine, and (2) the degree of inhibition by relatively high concentrations of cystine. The compositions of the virus populations in most of the passage levels were determined, and from these determinations the patterns of emergence of the mutant viruses were delineated.

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supporting

confidence: 48%

Section: Methodsmentioning

confidence: 99%

Poliovirus Mutants with Altered Responses to Cystine

Dubes

Chapin

1958

Journal of General Microbiology

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“…detection of virus by methods already described (9,10). Five tubes were used for each specimen, each being inoculated with 0.1 ml.…”

Section: Monl'~ey Kidney Cultures--trypsinized Rhesus Monkey Kidney mentioning

confidence: 99%

The Infection of Chimpanzees With Echo Viruses

Itoh

Melnick

1957

The Journal of Experimental Medicine

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The oral and parenteral infections of chimpanzees receiving echo Types 6 and 4 viruses successively are described. The two infections, spaced 2½ months apart, and given by the same route in each animal, failed to induce overt disease. The inapparent infections were demonstrated by virus excretion in the throat and the stools and the development of neutralizing antibodies. Complement-fixing antibodies also appeared after Type 6 infection, but fell more rapidly than the neutralizing antibodies. After oral infection, echo-6 virus was found for equal periods in both the throat and feces, but echo-4 persisted in the throat for much longer periods than in the lower bowel. Almost no virus carriage occurred after parenteral inoculation. No true viremia was exhibited in any of the animals. One of the chimpanzees had neutralizing antibodies against Type 6 virus in its pre-inoculation serum. It responded extraordinarily to the Type 6 exposure, developing antibody levels of 1:50,000 to echo-6, of 1:1024 against the echo-6' variant, and of 1:64 against the echo 6'' variant. Although Type 4 antibodies developed after the exposure, they proved difficult to measure by ordinary methods. However, they could be satisfactorily assayed by the plaque reduction method. Three other chimpanzees fed echo-2, echo-3, and an untypable echo virus, respectively, yielded results confirming those established with Types 4 and 6.

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“…* Table 11 shows a comparison of 4 16 animals developed disease, and this was also associated with lesions. As shown in the last three columns of the table, viremia was present on days two, three, and four, being a relatively common finding when a virulent strain was inoculated intramuscularly into monkeys.…”

mentioning

confidence: 99%

“…16 Parallel virus titrations were performed using 0.1 and 0.4 per cent bicarbonate. The low bicarbonate or acid cultures were kept in stoppered bottles or in loosely capped bottles in a 10 per cent CO.-incubator.…”

mentioning

confidence: 99%