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Biomedical subjects

M Onuma

Publications and source records attributed to M Onuma.

At least 253 records · Page 14Linked to original sources

Induction of transformed phenotypes in sheep fibroblasts by culture fluids from cells persistently infected with bovine leukemia virus.

FLK cells are fetal lamb kidney cells persistently infected with bovine leukemia virus (BLV). 3178 cells, originating from calf-form bovine lymphosarcoma, also showed persistent production of BLV and alteration of cell morphology, after treatment with 5'-iodo-2'-deoxyuridine. In the present paper, the first in vitro transformation of sheep fibroblasts by inoculation with BLV materials from these two cell lines is described. In a few passages after inoculation with these viral materials, morphological alteration occurred. The morphologically altered cells were grown as stable cultures and showed such transformed phenotypes as growth in soft agar medium, increased uptake of 2-deoxy-D-glucose and tumorigenicity in athymic nude mice. This result, together with our previous observation of simultaneous induction of BLV expression and morphological alteration of 3178 cells, suggests the presence of some transforming capacity in these BLV materials similar to that in, for example, murine or avian acute leukemia viruses. The possible acquisition of such capacity during the prolonged passage is discussed.

Animals↗

Transformed phenotypes in long-term cultures persistently infected with bovine leukemia virus.

Two cell lines, 3178 and FLK, were established respectively from calf form bovine lymphosarcoma and from fetal lamb kidney cells inoculated in vitro with bovine leukemia virus. These two cell lines persistently infected with bovine leukemia virus were maintained for more than 150 passages over three years. They exhibited the characteristics of transformed cell lines in vitro: 1) anchorage independence, 2) increased saturation density and decreased population doubling time, 3) increased uptake of 2-deoxy-D-glucose and 4) tumorigenicity in athymic nude mice.

Animals↗

Cohelical arrangement of the DNA strand in the core of Marek's disease virus particles.

Electron microscopic studies on Marek's disease virus demonstrated that fibrils of 20 to 30 A were arranged in a cohelical configuration within an electronopaque band of about 100 A thickness in the core of immature particles. These fibrils were presumed to be the DNA double helix itself. This configuration may represent an arrangement of the herpesvirus DNA.

Capsid↗

Cytotoxic antibody in cattle and sheep exposed to bovine leukemia virus.

Sera from bovine leukemia virus-infected cattle and sheep lysed fetal lamb kidney cells in the presence of rabbit complement. This cytolytic activity was removed completely from the sera by absorption with bovine leukemia virus (BLV). Antiserum against surface glycoprotein antigens of BLV contained cytolytic antibody but antiserum against the internal protein p24 did not. The complement-dependent antibody cytotoxicity test employing the trypan-blue dye exclusion method appeared to be suitable for routine diagnosis of BLV infection.

Animals↗

Antigenic difference between intracellular and membrane antigens induced by herpesvirus of turkeys.

The difference between intracellular antigen (IA) and membrane antigen (MA) induced by herpesvirus of turkeys (HVT) was examined by immunofluorescence analysis. Convalescent sera from chickens infected with HVT (HVT convalescent serum) and hyperimmune sera from chickens immunized with partially pruified virus (anti-HVT particle serum) or with a major HVT soluble precipitin antigen (anti-common Ag serum) were used as the source of antisera to specify both antigens in these experiments. Absorption of a convalescent serum from HVT-inoculated chicken with IA positive cells, resulted in reduction of their reactivity to IA but not to MA. Differential absorption of the hyperimmune sera by IA positive cells and MA positive cells resulted in reduction of their reactivity to homologous cells but not to heterlogous cells which were used in the absorptions. When the relationship between the levels of anti-IA and anti-MA antibodies was examined by a blocking immunofluorescence test using 38 sera from chickens inoculated with HVT, no correlation was observed (p greater than 0.25). These results provide evidence that MA and IA are antigenically different from each other.

Animals↗

Relation between the neutralization of herpesvirus of turkeys and the antibody to late-appearing membrane antigen induced by the virus.

The relation between the neutralization of herpesvirus of turkeys HVT) and the antibodies to early appearing membrane antigen (EMA), late appearing membrane antigen (LMA) and intracellular antigen (IA) induced by the virus was examined by blocking of direct immunofluorescence using 38 sera from chickens infected with HVT. Our results provide evidence that anti-LMA is significant in the neutralization of the infectivity (P less than 0.001) since the neutralizing capacity of the serum was related to its blocking index (BI) and not to its anti-EMA titre (BI). Furthermore, the anti-IA titres (BI) of the sera were also related to their neutralizing activity to a lesser extent (P less than 0.025). However, no relationships among any two of the titres of EMA, LMA and IA were observed. The implication of these results of the relationship of LMA to infectious virus particles is discussed.

Animals↗

Cell fusion activity of bovine leukaemia virus.

Bovine leukaemia virus (BLV) induces two types of cell fusion. Cells inoculated at high multiplicity show cell fusion within 2 h. This type of fusion can be induced by non-infectious virus and does not require protein synthesis for induction. Low m.o.i. also induces cell fusion but this depends on production of infectious virus.

Animals↗

Detection of bovine leukemia virus by syncytium assay.

When various indicator cells, including virus transformed and nontransformed cells, were cocultivated with bovine leukemia virus-producing cells, strong positive syncytia formation was found in transformed cells one day after cocultivation. The results of comparison of bovine leukemia virus antibody titers and the detection of bovine leukemia by the syncytium assay showed 89% of serologically positive cows were positive for bovine leukemia virus, whereas no reactors were found in serologically negative cows. However, the frequency of bovine leukemia virus detection differed according to the difference of incubation periods in the syncytium assay. Therefore, it is important to choose the appropriate indicator cell and culture conditions for the detection of bovine leukemia virus in the syncytium assay.

Animals↗