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Biomedical subjects

C R Parish

Publications and source records attributed to C R Parish.

At least 109 records · Page 6Linked to original sources
PubMed1984

The structural proteins of chick embryo lethal orphan virus (fowl adenovirus type 1).

Chick embryo lethal orphan (CELO) virus (fowl adenovirus type 1) contains at least 14 structural proteins with polypeptide molecular weights ranging from 100K to about 6K. A nomenclature of the CELO virion polypeptides is presented and the molar proportion of each polypeptide has been estimated. The CELO virus pentons were specifically released from the virion by dialysis against borate-based calcium-magnesium saline. The penton base (polypeptide III, mol. wt. 92K) and two fibres were separated, characterized and their polypeptides were correlated with their morphological positions in the virion. Peptide mapping suggested that the long fibre (polypeptide IV, mol. wt. 65K), and the short fibre (polypeptide VII, mol. wt. 44.5K) were not related in their primary sequences and are therefore probably encoded by separate genes. The time course of synthesis of the CELO virion polypeptides indicated that, like their mammalian adenovirus counterparts, they are synthesized late (after viral DNA replication).

Adenoviridae↗
PubMed1984

DNA-binding proteins of chick embryo lethal orphan virus: lack of complementation between early proteins of avian and human adenoviruses.

Chick cells infected by chick embryo lethal orphan (CELO) virus (fowl adenovirus type 1) contained four prominent virus-specific, structurally related DNA-binding proteins with mol. wt. of 74K, 64K, 56K, 52K, and two minor forms. The CELO virus DNA-binding proteins were phosphorylated, delayed-early nuclear proteins. CELO virus early proteins were expressed in BHK cells, but did not complement human adenovirus type 5 mutants with lesions E1A, E2A or E2B. Moreover, CELO virus DNA-binding proteins were not produced in 293 cells, which express human adenovirus E1 genes. These results suggest that activation of transcription by adenovirus E1A genes involves specific interactions between the E1A gene products and viral early promoters.

Adenoviridae↗
PubMed1984

Structural organization and polypeptide composition of the avian adenovirus core.

CELO virus (fowl adenovirus 1) contained three core polypeptides of molecular weights 20,000, 12,000, and 9,500. The core was similar to that of human adenoviruses, with some evidence of compact subcore domains. Micrococcal nuclease digestion of CELO virus cores produced a smear of DNA fragments of gradually decreasing size, with no nucleosome subunit or repeat pattern. Moreover, when digested cores were analyzed without protease treatment, there was again no evidence of a nucleosome substructure; neither DNA fragments nor core proteins entered a 4% polyacrylamide gel. The organization of the core is thus quite unlike that of chromatin. Restriction endonuclease analysis of the DNA from digested cores showed that the right end was on the outside of the core. We suggest that adenovirus DNA is condensed into the core by cross-linking and neutralization by the core proteins, beginning with the packaging sequence at the center of the core and ending with the right end of the DNA on the outside.

Adenoviridae↗
PubMed1984

Demonstration of lymphocyte surface lectins that recognize sulphated polysaccharides.

Cholate extracts of murine lymphocytes were shown to contain haemagglutinating activity against autologous erythrocytes. The species specificity and sugar inhibition pattern of the haemagglutinin closely paralleled the specificity of autorosetting, an interaction that had been shown previously to involve the recognition by lymphocytes of carbohydrate structures on autologous erythrocytes. The probable identity of the haemagglutinin and autorosetting receptors was confirmed by experiments utilizing the unique plasma protein autorosette inhibition factor, which appears to block both interactions by masking carbohydrate acceptor sites on erythrocytes. Detailed sugar inhibition studies revealed that the haemagglutinin and autorosetting receptors have a high affinity for certain sulphated polysaccharides, such as heparin and dextran sulphate. Since similar sulphated polysaccharides have been shown previously to inhibit lymphocyte recirculation, a possible role for these receptors in lymphocyte homing and recirculation is discussed.

Agglutinins↗
PubMed1983

A rapid, automated colorimetric assay for measuring antibody binding to cell surface antigens.

An automated, colorimetric procedure is described for detecting antibodies specific for cell surface antigens. The procedure entails (a) coating the wells of 96-well microplates with either protein A or anti-immunoglobulin antibodies and (b) preincubating either the microplate or target cells with the test antibody. Target cells which react with the test antibody bind to the wells of the microplate and bound cells are quantitated by staining with the dye Rose Bengal. A microplate spectrophotometer is used to measure absorbance in each well of the plate, providing a rapid, automated measure of antibody titre. The assay is simple to perform, uses readily available reagents and gives comparable sensitivity to rosetting assays. With these features, and the capacity for handling large numbers of trays quickly, this method has obvious advantages in screening for antibody activity in culture supernatants of hybridoma clones.

Animals↗
PubMed1983

Automated colorimetric assay for T cell cytotoxicity.

A colorimetric method has been developed for detecting the lysis of target cells by cytotoxic T lymphocytes (Tc). The method entails incubating Tc cells with thioglycollate-induced macrophage targets and estimating macrophage survival at the end of the assay by staining viable macrophages with the dye neutral red. The method is substantially more sensitive than the 51Cr release assay and can be used to detect alloreactive Tc cells and H-2-restricted Tc cells against viruses, haptens and minor-H antigens. Furthermore, the assay is applicable to limit dilution analysis of Tc cell precursors. The method is cheap, avoids radioactive materials and by measuring optical densities with automated spectrophotometers developed for microELISA systems, results can be obtained 50-100 times faster than with the radioactive procedure.

Animals↗
PubMed1983

A comparison of the terminal protein and hexon polypeptides of avian and human adenoviruses.

It was found that the virion terminal protein of chick embryo lethal orphan (CELO) virus had a molecular weight of 46,000, and the hexon a molecular weight of 100,000. 125I-labelled tryptic and chymotryptic peptide maps of the hexons and terminal proteins from CELO virus and human adenovirus type 5 (Ad5) differed. However, limited proteolysis of CELO virus and Ad5 terminal proteins by protease V8 showed similarities which were not detected in the case of the two hexons.

Adenoviridae↗
PubMed1983

Adenovirus-induced alterations of the cell growth cycle: a requirement for expression of E1A but not of E1B.

Mutants dl312, dl314, hr1, and hr3 with mutations in region E1A of adenovirus type 5 were defective for the induction of cell cycle abnormalities detectable by flow cytometry, cell DNA replication, thymidine kinase production, and chromosome aberrations and did not synthesize the viral DNA-binding protein (E2A) in rat cells. dl311, a leaky E1A mutant, induced cell cycle effects at high multiplicity in only one of three experiments, and synthesized the DNA-binding protein. hr7 (E1B) gave a wild-type response in all tests. dl313 was also positive in all tests, although it induced fewer polyploid cells than did wild-type virus, probably because of the leftward extension of the dl313 E1B deletion into E1A. sub315 and sub316, with mutations which also span the E1A-E1B border, synthesized DNA-binding protein, but caused no cell cycle alterations detectable by flow cytometry in rat or mouse cells. Although the participation of other viral early regions cannot be completely excluded, our results suggest that alteration of cell cycle progression is a direct effect of E1A unrelated to its control of other viral early regions, and may be the function of E1A in transformation.

Adenoviruses, Human↗
PubMed1982

Automated rosetting assay for cell surface antigens and autorosetting lymphocytes.

An automated procedure, using a Coulter counter, is described for enumerating rosette-forming lymphocytes in 2 rosetting systems in mice, detecting antibodies to cell surface antigens, and the interaction of autologous erythrocytes with thymocytes (autorosetting). The procedure gives results comparable with determinations of rosettes by light microscopy. The procedure not only estimates rosetting percentages, but can be used to titrate anti-lymphocyte antibodies, to detect autorosette inhibition factor in serum and to assay cell surface antigens in detergent lysates of spleen cells.

Animals↗
PubMed1982

Latex bead rosetting method for cell surface antigens.

Polystyrene latex beads coupled with anti-Ig can be used to detect the binding of specific antibodies to target cells. The optimal conditions for coupling anti-Ig to beads are described. This method is as sensitive as red cell rosetting and has the advantages that coupling is simple and the coated beads stable.

Animals↗
PubMed1982

Inhibition of secondary IgG responses by monosaccharides: evidence for I-region control.

Certain monosaccharides selectively inhibit secondary IgG responses in vitro. Genetic analyses described in this report revealed that the inhibitory sugars differed between mouse strains and these differences mapped to the I-J and I-C subregions of the murine MHC. These results imply that interaction between T and B lymphocytes can involve the recognition of I-region controlled carbohydrate structures.

Acetylgalactosamine↗
PubMed1982

Anti-self receptors. V. Properties of a mouse serum factor that blocks autorosetting receptors on lymphocytes.

Murine lymphocytes spontaneously bind autologous and allogeneic erythrocytes via receptors that primarily recognize self H-2L molecules on the erythrocyte surface. Normal mouse serum contains a factor, termed autorosette inhibition factor (AIF), that very effectively blocks autorosette formation. This paper describes experiments that determine the origin and nature of serum AIF. It was found that AIF lacks strain and species specificity, serum from several mammalian and non-mammalian species inhibiting the autorosetting of BALB/c thymocytes. However, mouse strains differed in the levels of AIF in their serum. Furthermore, AIF appears to directly interact with autorosetting receptors on lymphocytes as thymocytes from the BALB/c-H-2dm2 mutant strain, which lack autorosetting receptors, were unable to absorb the factor. Several lines of experimental evidence indicated that AIF is secreted by a population of short-lived, radiosensitive macrophages (or monocytes). Firstly, in vivo administration of the anti-macrophage agents carrageenan and silica profoundly depressed AIF levels in serum. Secondly, in vitro culturing of different lymphoid cells revealed that AIF is secreted by an adherent population of peritoneal cells. Thirdly, total body irradiation experiments demonstrated that AIF production is dependent upon a radiosensitive cell that is bone marrow derived. Finally, AIF was purified to homogeneity from mouse plasma and shown to be a single polypeptide chain with a molecular weight of 84,000.

Aging↗
PubMed1981

Anti-self receptors. IV. H-2-restricted receptors on thymocytes recognize carbohydrate structures on target cells.

Rosetting between thymocytes and autologous erythrocytes in mediated by receptors on thymocytes that primarily recognize self H-2L molecules on erythrocytes. This paper describes preliminary attempts to chemically characterize the receptor and acceptor molecules involved in this H-2-restricted interaction. On the basis of sugar inhibition studies and the sensitivity of the receptors to protease and glycosidase treatments it appears that a protein receptor on thymocytes recognizes the carbohydrate portion of a glycoprotein on erythrocytes. Furthermore, the thymocyte receptor appears to recognize terminal D-galactose, D-mannose and sialic acid residues on a branched-chain carbohydrate structure on erythrocytes, with mouse strains of different H-2 haplotype expressing carbohydrate structures that differ in the linkage of these three terminal sugars. These findings indicate that H-2-restricted carbohydrate-protein interactions can occur between cells, a conclusion with important theoretical implications.

Animals↗
PubMed1981

Inhibition of secondary IgG responses by N-acetyl-D-galactosamine.

A range of monosaccharides were tested for their ability to inhibit a variety of in vitro immune response. The most striking specific inhibition was produced by N-acetyl-D-galactosamine (GalNAc). This sugar strongly inhibited the secondary IgG antibody response to two different hapten-carrier systems, but had no effect on primary and secondary IgM responses, generation of cytotoxic T cells to alloantigens and mixed lymphocyte reactions. By exposing secondary antibody cultures to GalNAc for varying periods of time, it was observed that GalNAc only exerted its inhibitory effect on day 4 of the culture, the day when IgG plaque-forming cells first appeared. Furthermore, GalNAc could override the action of T helper factor in T cell-depleted cultures. Collectively, these data indicate that GalNAc inhibits the initiation of IgG synthesis probably by blocking the interaction of a helper factor for IgG synthesis with its target cell.

Acetylgalactosamine↗
PubMed1981

H-2U: a new region at the D end of the murine MHC.

A new genetic region, mapping within the H-2 complex, has been serologically defined with several alloantisera raised in mice which differ at the D region. When these antisera were absorbed to remove H-2D antibodies, residual antibody activity remained that reacted in a strain-specific manner, and the antigens involved mapped to a new genetic region between the S and D regions. Two allelic variants relating to the d and k haplotypes have been defined by genetic mapping studies. This new region has been designated H-2U and the antigens it controls appear to resemble Ia antigens in their cellular distribution and molecular weight. The new antigen is primarily expressed on B cells, and is carried on protein molecules having approximate molecular weights of 36 000 and 60 000 daltons and resembling the alpha and alpha--beta chain dimer characteristic of Ia antigens.

Animals↗