PubMed Health⌕ Search

Biomedical subjects

V G Chinchar

Publications and source records attributed to V G Chinchar.

44 records · Page 3Linked to original sources

Localization of frog virus 3 proteins using monoclonal antibodies.

Thirty-seven monoclonal antibodies to seven frog virus 3 (FV3) structural proteins were isolated and used to examine the distribution of viral proteins within virions and infected cells. Three monoclonal antibodies, one to the major capsid protein, VP55, and two to VP38 had detectable neutralizing activity suggesting that these proteins are located on the surface of virions. Immunofluorescent studies showed that VP108, VP57, VP55, and VP16 were localized mainly within virus assembly sites, while VP17 was detected in both assembly sites and the surrounding cytoplasm. The abundance of viral structural proteins within assembly sites is consistent with the idea that virion maturation occurs exclusively within assembly sites.

Animals↗

Early proteins are required for the formation of frog virus 3 assembly sites.

The formation of frog virus 3 virions takes place within morphologically distinct regions of the cytoplasm termed assembly sites. These sites are formed within infected BHK cells by 6-7 hr after infection, a time when viral DNA and both early and late proteins are present. To identify macromolecules involved in assembly site formation, a temperature-sensitive mutant ( ts9467 ) was used which is not only defective in the synthesis of late RNA and proteins (D.B. Willis, R. Goorha , and A. Granoff , 1979, Virology 98, 328-335), but, as reported here, also does not form assembly sites at nonpermissive temperatures. When ts9467 -infected cells were shifted from the nonpermissive to permissive temperature, assembly sites were observed within 1 hr even when late protein synthesis was inhibited by cycloheximide. Monoclonal antibodies specific for early and late viral proteins were used to show that assembly sites formed under these conditions contained at least one early protein, but lacked four representative late proteins. These results indicate that assembly site formation involves interaction between one or more early proteins and viral DNA, and that late proteins do not play a role in this process.

Animals↗

Characterization of hybrids between bovine (MDBK) and mouse (L-cell) cell lines.

Hypoxanthine-guanine phosphoribosyltransferase (HGPRT)-deficient mutants of a bovine kidney cell line (MDBK) were selected following mutagenesis with ethylmethane sulfonate or ICR-170G. MDBK mutants were hybridized to thymidine kinase-deficient L cells and selected in HAT medium. Parental and hybrid cells were characterized for isozyme patterns of lactic dehydrogenase malate dehydrogenase, glucose-6-phosphate dehydrogenase, and glutamate oxalate transaminase. Chromosomes of MDBK can be distinguished from mouse L cells by configuration and by fluorescent staining with Hoechst 33-258 stain. Hybrid cells contained both MDBK and L-cell chromosomes and had elevated DNA content. MDBK cells are normally restrictive for mengovirus replication. Both permissive and restrictive hybrids were found. Our data indicate that there was preferential loss of MDBK chromosomes in the hybrid cell lines.

Animals↗

Anti-viral cytotoxic cells in the channel catfish (Ictalurus punctatus).

Cytotoxic cells isolated from the head kidney and peripheral blood of the channel catfish appear to represent distinct subpopulations of effector cells. Previous studies showed that the former lyse xenogeneic natural killer (NK) cell targets, whereas the latter preferentially lyse allogeneic cells. Here we extend these studies and present data suggesting a third class of cytotoxic effectors responsible for killing virus-infected allogeneic and autologous cells. Peripheral blood leukocytes (PBLs) freshly isolated from unimmunized catfish lyse uninfected allogeneic target cells as well as virus-infected allogeneic and autologous cells. Cell depletion and unlabeled ("cold") target inhibition studies discriminated between putative effector classes and supported the view that at least two populations of cytotoxic cells are present within peripheral blood leukocytes. One population lyses allogeneic targets, whereas a second population kills channel catfish virus (CCV)-infected cells. In addition, inhibitor studies demonstrated that early virus gene products are sufficient to render infected cells susceptible to lysis. These results suggest that channel catfish possess distinct populations of NK-like, PBL-derived cytotoxic cells capable of lysing allogeneic and virus-infected target cells.

Acyclovir↗

MHC class II B genes in the channel catfish (Ictalurus punctatus).

Two different cDNA sequences for major histocompatibility complex (MHC) class II beta chains from the channel catfish (Ictalurus punctatus) have been identified. Homology between these sequences and those previously identified as MHC class II B genes in other teleosts suggests they represent alleles of the DAB locus. The inferred amino acid sequences show strong evidence for a functional polypeptide chain with a peptide binding region. Southern blot analysis reveals polymorphism in the MHC class II B gene(s) of the channel catfish and suggests the presence of two to four genes.

Amino Acid Sequence↗