A morphologic comparison of tumors produced by type 12 adenovirus and by the HA-12-1T line of adeno-12 tumor cells.
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Biomedical subjects
Publications and source records attributed to D S Yohn.
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A significantly higher proportion of female hamsters developed tumors than did males given the same dose of adenovirus-12 (Huie) at birth over a dose range from 10(5.0) to 10(7.0)tcd(50) for human embryonic kidney cells. The 50% tumor dose (td(50)) was calculated to be 10(5.90)tcd(50) for females and 10(6.27) for males. Tumor response patterns induced with approximate td(50) inocula, 10(6.0) for females and 10(6.3) for males, were quite similar. The greater susceptibility of females was not found to be characteristic of a single strain of hamsters; nor was it attributable to a single lot of virus, to a single type of human cell used to produce the virus, nor to the degree of purification of the virus inoculum. The inoculation route did not appear to be of importance. Inasmuch as the foregoing extrinsic factors were of little influence, it was concluded that the mechanism is host-mediated, presumably hormonally controlled. The possibility that female cells, independent of host control, are more susceptible to adenovirus-12 oncogenesis than male cells has not been explored. Tumor regression occurred in 20% of the 211 tumors in males and in 15% of the 355 tumors in females. Adenovirus-12 T-antibody was detected in all but six of 473 sera tested from tumor-bearing hamsters and in 50% of 94 sera tested from non-tumor-bearing animals given virus at birth. Antibodies in the latter group were detected almost exclusively by indirect immunofluorescence. This technique appears to be extremely sensitive for detection of low levels of adeno-12 T-antibodies. The implications of T-antibody in nontumor-bearing virus-injected hamsters are discussed. Sera from normal hamsters were free of T-antibody.
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Yohn, David S. (Roswell Park Memorial Institute, Buffalo, N.Y.), Fanny R. Marmol, Victoria A. Haendiges, and James T. Grace, Jr. Yaba tumor poxvirus synthesis in vitro. II. Adsorption, inactivation, and assay studies. J. Bacteriol. 91:1953-1958. 1966.-Means to increase the efficiency of the Yaba tumor poxvirus assay in BSC-1 cell cultures were sought. A method was devised wherein 0.4 ml of each virus dilution was layered onto BSC-1 cells in 80-mm Leighton tubes and permitted to adsorb at 25 C for 18 hr prior to incubation at 35 C. The diluent found most reliable was medium 199 containing 50% bovine amniotic fluid adjusted to 2.0 mm calcium and 1.0 mm magnesium at pH 7.0. These modifications yielded highly reproducible titrations with a greater than twofold increase in assay sensitivity. Irreversible adsorption of Yaba virus by BSC-1 cells proceeds comparatively slowly at 25 to 33 C. Although the process is more rapid at 35 or 37 C, the increased thermal lability of the virus at these temperatures results in lower titers than with virus adsorbed at 25 C for longer periods of time. Yaba poxvirus appears to be 5 to 10 times more sensitive to thermal inactivation than vaccinia poxvirus.
Yohn, David S. (Roswell Park Memorial Institute, Buffalo, N.Y.), Victoria A. Haendiges, and James T. Grace, Jr. Yaba tumor poxvirus synthesis in vitro. I. Cytopathological, histochemical, and immunofluorescent studies. J. Bacteriol. 91:1977-1985. 1966.-Yaba tumor poxvirus synthesis in BSC-1 cell culture was followed sequentially with light microscopy, immunofluorescent microscopy, and various histochemical stains. The first evidence of infection was detected at 24 hr when nucleoli became hypertrophic, reflecting enhanced ribonucleic acid (RNA) synthesis. At 36 hr, deoxyribonucleic acid synthesis was detected in the cytoplasm. This was immediately followed by or associated with antigen synthesis at paranuclear sites and enhanced RNA synthesis in the cytoplasm. Cytoplasmic inclusions were readily apparent at 4 days in initially infected cells. Contiguous spread of virus was judged to have occurred around the third day of infection. The time required to complete the synthetic cycle from time of infection to production of infectious progeny was estimated to be of the order of 60 hr. This cycle is 6 to 10 times longer than for vaccinia virus. By light microscopy, cytopathic effects (CPE) were detectable in 5 days in heavily infected cultures. With 100 units or less of infectious virus, CPE was not readily detected until the 10th to 14th day. At this time, focal areas of infection classified either as microtumors or microplaques were present. Secondary foci appeared during the 4th week of incubation.
Yohn, David S. (Roswell Park Memorial Institute, Buffalo, N.Y.), Victoria A. Haendiges, and Etienne de Harven. Yaba tumor poxvirus synthesis in vitro. III. Growth kinetics. J. Bacteriol. 91:1986-1991. 1966.-The infectious synthetic cycle of Yaba poxvirus by BSC-1 cells at 35 C is described and discussed together with related observations obtained by immunofluorescence and electron microscopy. After adsorption of virus at 25 C, at least 6 hr of incubation at 35 C was required before viral eclipse was demonstrable. Infectious progeny appeared after an additional 48 hr of incubation at 35 C. Newly synthesized virus particles were seen at 68 hr but not at 44 hr by electron microscopy. Maximal maturation occurred between 100 and 120 hr. Maximal yields were obtained from the 5th through the 8th day. The differences in kinetics between Yaba and vaccinia poxviruses, as described for the latter in the literature, are discussed. Possible factors accounting for these differences are enumerated.
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