PubMed Health⌕ Search

Biomedical subjects

J C Glorioso

Publications and source records attributed to J C Glorioso.

At least 163 records · Page 9Linked to original sources
PubMed1983

Role of interferon in human natural killer activity against target cells infected with HSV-1.

Human natural killer (NK) cells show high cytotoxic activity against target cells infected with herpes simplex virus type 1 (HSV-1). Substantial amounts of interferon (IFN) were generated in co-cultures of NK effector cells and infected target cells; however, the cytotoxic activity seen against a specific infected cell target did not correlate with the amount of IFN induced. The production of IFN increased steadily from 4 to 18 hr of co-culture, as did NK activity; however, IFN production peaked 4 hr later than NK activity. Pretreatment of NK effector cells with exogenous IFN increased cytotoxic activity against all targets tested, but the differential pattern of reactivity against cells infected with wild type and mutant viruses was unaltered. When effector cells were treated with the RNA synthesis inhibitor actinomycin D before co-culture with virus-infected targets, IFN production was markedly reduced, without a concomitant reduction in cytotoxicity. Similarly, the addition of anti-IFN antiserum to co-cultures greatly decreased the available IFN present, but had no effect on NK activity. We conclude that the induction of cytotoxic activity in co-cultures of NK effector cells and HSV-1-infected target cells is independent of the induction of IFN.

Adult↗
PubMed1982

Adhesion of type A Pasteurella mulocida to rabbit pharyngeal cells and its possible role in rabbit respiratory tract infections.

Pasteurella multocida serotype A was found in association with the mucosal epithelium of the nasopharynges of rabbits with respiratory tract infections. The bacteria specifically attached to squamous epithelial cells of the pharyngeal mucosa both in vivo and in vitro and to some tissue culture cell lines such as HeLa. All strains with serotype A capsules were adhesive. With the exception of one serotype D strain, strains with capsular serotypes B, D, and E were at least 10-fold less adhesive. Bacterial adhesiveness was much reduced after pronase digestion, heat treatment, and homogenization, but removal of the hyaluronic acid capsule increased adhesion. Electron microscopy revealed that fimbriae were produced by an adhesive pasteurella strain, but not by two nonadherent strains. The attachment of the former strain to pharyngeal and HeLa cells was inhibited by N-acetyl-D-glucosamine. Together, these findings suggest that this amino sugar may be a component of the receptor on both animal cell surfaces and that the fimbriae may be the adhesions. It is proposed that bacterial attachment has a role in colonization and infection of rabbit upper respiratory mucosae.

Adhesiveness↗
PubMed1981

Method for induction of mutations in physically defined regions of the herpes simplex virus genome.

A procedure was developed for inducing mutations in isolated restriction enzyme fragments of herpes simplex virus type 1 (HSV-1) DNA with nitrous acid. The mutations were then transferred to the viral genome by genetic recombination during cotransfection of rabbit kidney cells with the mutagenized fragments and intact HSV-1 DNA. The HpaI restriction enzyme fragments LD, B, LG, I, and J were mutagenized. Temperature-sensitive mutants were found at frequencies of 1 to 5% among the progeny of the transfections. Syncytial mutants also were found at high frequency when fragment B or LD was used for mutagenesis. Fifteen of these mutants, 11 temperature sensitive and 4 syncytial, were used for further studies, including complementation analysis, DNA synthesis, and marker rescue. Marker rescue data presented here and in the accompanying publication (A. L. Goldin, R. M. Sandri-Goldin, M. Levine, and J. C. Glorioso, J. Virol. 38: 50-58, 1981) confirm the map position of some of the newly isolated mutants.

DNA Restriction Enzymes↗
PubMed1981

Cloning of herpes simplex virus type 1 sequences representing the whole genome.

Sequences representative of the whole genome of herpes simplex virus type 1 (HSV-1) strain KOS were cloned in the plasmid vector pBR325 in the form of EcoRI-generated DNA fragments. The cloned fragments were identified by digestion of the chimeric plasmid DNA with restriction enzymes EcoRI or EcoRI and BglII followed by comparison of their electrophoretic mobilities in agarose gels with that of similarly digested HSV-1 virion DNA. The cloned fragments showed the same migration patterns as the corresponding fragments from restricted virion DNA, indicating that no major insertions or deletions were present. The presence of HSV-1 sequences in the chimeric plasmids was confirmed by hybridization of plasmid DNA to HSV-1 virion DNA. Additionally, some of the cloned fragments were shown to be biologicaly active in that they efficiently rescued three HSV-1 temperature-sensitive mutants in cotransfection marker rescue experiments.

Cloning, Molecular↗
PubMed1981

High-frequency transfer of cloned herpes simplex virus type 1 sequences to mammalian cells by protoplast fusion.

The protoplast fusion technique of Schaffner (W. Schaffner, Proc. Natl. Acad. Sci. U.S.A. 77:2163-2167, 1980) has been adapted to introduce cloned herpes simplex virus genes into cultured mammalian cells. The technique involves digesting bacterial cell walls with lysozyme to produce protoplasts and then fusing the protoplasts to mammalian cells by treatment with polyethylene glycol. For monitoring transfer, protoplasts were labeled with the fluorescent dye fluorescein isothiocyanate before fusion. After fusion, greater than 50% of the mammalian cells were fluorescent, demonstrating that bacterial material was transferred with high frequency. Transfer of plasmid pBR325 occurred at frequencies of 1 to 2%, as measured by in situ hybridization. Fusion transfer of a chimeric plasmid consisting of the herpes simplex virus type 1 (strain KOS) EcoRI fragment F in pBR325 resulted in expression of some viral genomic sequences in about 5% of the mammalian cells, as detected by indirect immunofluorescence. One Ltk- cell in 300 to 500 was transformed to the TK+ phenotype after fusion with protoplasts carrying the chimeric plasmid pX1, which consists of pBR322 and the BamHI fragment coding for the herpes simplex virus type 1 thymidine kinase gene.

Animals↗
PubMed1981

Enzyme-linked immunosorbent assay of serum protein SAA in rhesus monkeys with secondary amyloidosis.

Tissue deposits of amyloid protein AA and a concomitant elevation of serum protein SAA have been demonstrated previously in mice and humans with secondary amyloidosis, but not in rhesus monkeys (Macaca mulatta). In this study, protein SAA was quantitated in normal and amyloidotic rhesus monkeys using an enzyme-linked immunosorbent assay. Protein AA was isolated from the liver of a rhesus monkey with secondary amyloidosis by a combination of water extraction and gel filtration chromatography. The purified material had rigid, nonbranching fibrillar structures typical of amyloid on electron microscopy and a molecular weight of 9000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The amino acid content and partial sequence were comparable to those reported previously for protein AA of rhesus monkeys. As measured by enzyme-linked immunosorbent assay, normal rhesus monkeys had SAA levels of 40 to 64 ng. of AA equivalents per ml., whereas amyloidotic rhesus monkeys had SAA levels of 1700 to 95,000 ng. of AA equivalents per ml. An elevation of protein SAA was also detected in an amyloidotic pigtailed macaque (Macaca nemestrina) using rabbit antirhesus protein AA. Rabbit antisera against human and mouse protein AA reacted strongly with rhesus protein AA and with amyloidotic rhesus serum, but only slightly with normal rhesus serum, indicating that rhesus proteins AA and SAA have antigenic determinants cross-reactive with protein AA of xenotypic species.

Amino Acid Sequence↗
PubMed1981

Pasteurella associated rhinitis of rabbits: efficacy of penicillin therapy.

Thirty adult New Zealand white rabbits with chronic rhinitis were obtained from a commercial breeding colony. Penicillin sensitive strains of Pasteurella multocida were isolated from the upper respiratory tract of 28 (93%) of these rabbits. The diseased rabbits were treated with either intramuscular penicillin or penicillin nasal spray for 10 days and monitored for clinical signs of rhinitis and for the presence of Pasteurella multocida in the nasal passages. Rabbits receiving penicillin therapy by either route showed significant remission of the clinical signs of rhinitis during the study period; however, following treatment there was not significant difference in the prevalence of rhinitis between the treated groups and the untreated group. This was due in part to the considerable but non-significant improvement shown by the untreated group. This improvement which was not due to penicillin therapy may have been due to stabilization of environmental factors. The prevalence of Pasteurella multocida in the upper respiratory tracts of either the treated or untreated rabbits did not change significantly during the study period.

Animals↗
PubMed1981

Resistance of Pasteurella multocida to rabbit neutrophil phagocytosis and killing.

The susceptibilities of several Pasteurella multocida strains to serum bactericidins and to killing by rabbit polymorphonuclear neutrophils (PMN) were investigated, using in vitro assays. Strain Bunia II (serotype 5,12:E) was killed by both immune and normal rabbit sera containing active complement, and strains R1 (serotype 3,12:A) and 656 (serotype 12:B) were serum resistant. Strain R1 opsonized with immune or normal rabbit serum with complement or with immune antibody alone was phagocytized by neutrophils, and strain 656 was ingested only after opsonization with immune antibody in the presence or absence of complement. Complement alone was ineffective as an opsonin for the last 2 serotypes. Neither isolate was resistant to PMN killing. Growth of strain R1 in subcutaneously implanted chambers in rabbits did not increase the resistance of this organism to PMN. Comparison of phagocytosis and killing of this isolate with 2 virulent rabbit strains of P multocida--strains L-A (serotype 12:A) and 7228 (serotype 14:D)--demonstrated that strain L-A was resistant to destruction by neutrophils, whereas strains 7228 and R1 were killed. Resistance of strain L-A was not associated with the hyaluronic acid capsule.

Animals↗
PubMed1980

Persistence of herpes simplex virus genes in cells of neuronal origin.

The growth characteristics of the KOS strain of herpes simplex virus type 1 (HSV-1) in cell lines of nervous tissues origin were examined in an attempt to develop a tissue culture system mimicking the in vivo state of HSV-1 latency. We have previously reported that the B103 rat brain neuroma cell line is nonpermissive for growth of the KOS strain. In this report, we show that this nonpermissiveness is a temperature- and multiplicity-dependent phenomenon, with minimum virus yields at an elevated temperature and a low multiplicity of infection. Under these conditions, B103 cells survived infection with active wild-type or mutant HSV-1, whereas similarly treated Vero cells were killed. Six independent cultures of B103 cells surviving HSV-1 infection have been established. The surviving cells ceased production of any HSV-1 virus by 14 days postinfection and resumed growth and division at rates comparable to those of uninfected B103 cells. Survivor cells continued to express HSV-1-specific antigens, however, as detected by indirect immunofluorescence and by surface iodination followed by immunoprecipitation and polyacrylamide gel electrophoresis. The survivor cells did not express all of the surface proteins seen on productively infected B103 cells, and they were not susceptible to complement-mediated immune cytolysis with anti-HSV-1 antiserum. These results demonstrate that at least a portion of the HSV-1 genome is being harbored in these survivor cells.

Animals↗
PubMed1980

Mutant analysis of herpes simplex virus-induced cell surface antigens: resistance to complement-mediated immune cytolysis.

BHK-21 cells infected with temperature-sensitive mutants of herpes simplex virus type 1 strain KOS representing 16 complementation groups were tested for susceptibility to complement-mediated immune cytolysis at permissive (34 degrees C) and nonpermissive (39 degrees C) temperatures. Only cells infected by mutants in complementation group E were resistant to immune cytolysis in a temperature-sensitive manner compared with wild-type infections. The expression of group E mutant cell surface antigens during infections at 34 and 39 degrees C was characterized by a combination of cell surface radioiodination, specific immunoprecipitation, and gel electrophoretic analysis of immunoprecipitates. Resistance to immune lysis at 39 degrees C correlated with the absence of viral antigens exposed at the cell surface. Intrinsic radiolabeling of group E mutant infections with [14C]glucosamine revealed that normal glycoproteins were produced at 34 degrees C but none were synthesized at 39 degrees C. The effect of 2-deoxy-D-glucose on glycosylation of group E mutants at 39 degrees C suggested that the viral glycoprotein precursors were not synthesized. The complementation group E mutants failed to complement herpes simplex virus type 1 mutants isolated by other workers. These included the group B mutants of strain KOS, the temperature-sensitive group D mutants of strain 17, and the LB2 mutant of strain HFEM. These mutants should be considered members of herpes simplex virus type 1 complementation group 1.2, in keeping with the new herpes simplex virus type 1 nomenclature.

Animals↗
PubMed1979

In vitro killing of Pasteurella multocida: the effect of rabbit granulocyte and specific antibody source.

In vitro granulocyte-killing assays were performed to examine the ability of granulocytes from pasteurella-free or immunized rabbits, the in combination with specific immune serum, to kill Pasteurella multocida. Granulocytes from healthy rabbits and from rabbits with P multocida infections were equally competent. Granulocyte source, serum source, and specific antibody titer had no effect on granulocyte phagocytic activity. Moreover, serum containing specific antibody and complement supported the growth of the bacterium. These data suggest that chronic P multocida infections are not attributable to defective granulocytes or lack of serum antibody production.

Animals↗
PubMed1978

Complement receptors: specific detection by molecular complexes.

Immune complexes of BSA-anti BSA (IgG F(ab')2) will activate human complement by the alternate pathway. The C3 bound to these complexes binds to human peripheral blood mononuclear cells which bear complement receptors. This interaction is visualized by fluoresceinated antigen (BSA-FITC) or fluorescein conjugated antisera directed at the C3 component of complement. The assay appears to be more sensitive than the usual IgMEAC rosette technique and correlates well with the rosette method.

Animals↗
PubMed1978

Infection by herpes simplex virus and cells of nervous system origin: characterization of a non-permissive interaction.

Replication of herpes simplex virus type I (HSV-I) was studied in various cell lines of rat nervous system origin. Infection of neonatal rat glial primary cells with HSV-I, strain KOS, produced normal yields of progeny virus. Glioma lines B9 and B15 were permissive, the neuronal line B50 was partially restricted (10 to 100-fold reduction) and the neuronal line B103 was non-permissive (greater than 1000-fold reduction) for HSV-I (KOS) replication. Synthesis of virus DNA in infected B103 cells was not detected. However, at least some virus macromolecular synthesis was induced, including production of thymidine kinase, DNA polymerase and virus structural proteins.

Adsorption↗
PubMed1978

Complement-mediated cytolysis of HSV-1 and HSV-2 infected cells: plasma membrane antigens reactive with type-specific and cross-reactive antibody.

Surface antigens of BHK-21 cells infected with HSV-1 or HSV-2 were radioiodinated (125I) with lactoperoxidase, immune precipitated and analysed by polyacrylamide gel electrophoresis (PAGE). Experiments using antiserum to HSV-1 or HSV-2, absorbed with appropriate hemotypic or heterotypic antigens, revealed that both type-specific (homotypic) and cross-reactive antibody combined with surface glycoproteins to form a single large radioactive peak. This peak, which constituted the major glycoprotein region (region a) observed in electropherograms, represented a range in mol. wt. from 115000 to 130000. Sensitization of cells to complement lysis, neutralization of infectious virus and immune precipitation of surface glycoproteins (region a) were found to be generally correlated properties of all the antibody preparations analysed, including antibody prepared specifically against region a antigens. These findings suggest a major immunological role for the surface glycoproteins migrating in PAGE region a.

Animals↗
PubMed1978

Possible role of Fc receptors on cells infected and transformed by herpesvirus: escape from immune cytolysis.

Receptors for the Fc portion of nonimmune immunoglobulin G were demonstrated on B103 rat brain neuroma cells infected with herpes simplex virus type 1 (HSV-1) KOS by a radioimmunoassay using 125I-labeled heat-aggregated Fc fragments. Immune F(ab')2 fragments specific for HSV antigens competed efficiently for Fc binding sites, suggesting that the binding of Fc fragments to infected cells is specific for viral cell-surface antigens. It has been suggested that the binding of immune complexes to Fc receptors on the surfaces of tumor cells in vivo plays a role in protecting these cells from immune destruction. In vitro evidence is presented for the ability of aggregated immunoglobulin G molecules bound to cell-surface Fc receptors to protect both HSV-infected and HSV-transformed cells against complement-dependent and cell-mediated immune lysis.

Animals↗
PubMed1977

Immune interactions with cells infected with herpes simplex virus: antibodies to radioiodinated surface antigens.

Lactoperoxidase-catalyzed radioiodination was used to study reactions between surface antigens and antibodies on BHK-21 cells infected with HSV-1 and HSV-2. Isolation of iodinated surface antigens was achieved by indirect immune precipitation of Triton X-100 disrupted cells with antisera to HSV and IgG. Analysis of immune precipitates by polyacrylamide gel electrophoresis (PAGE) revealed at least 10 antigens, ranging in m.w. from 35 x 103 to 160 x 103 daltons. Antigens were detectable on cell surfaces as early as 2 hr post-infection. Electrophoretic patterns of surface antigens induced by HSV-1 were similar to those induced by HSV-2. In both cases the major portion of activity was associated with glycoprotein(s) in the range of 115 x 103 to 130 x 103 daltons. A reduced amount of radioactivity was obtained if cells were reacted with anti-HSV sera before disruption with Triton X-100, suggesting that less surface antigen was accessible to HSV antibody applied directly to intact cells.

Antibodies, Viral↗