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B T Rouse

Publications and source records attributed to B T Rouse.

At least 181 records · Page 10Linked to original sources
PubMed1984

Role of T-lymphocytes in the pathogenesis of herpetic stromal keratitis.

Our study was designed to investigate the mechanism of the stromal reaction in experimental ocular infection of murine eyes with herpes simplex virus (HSV). Severe stromal keratitis with scarring occurred in BALB/c mice after infection of the scarified cornea but similar reactions did not occur in athymic mice. However, if athymic mice were given adoptive transfers of lymphoid cells, a severe necrotizing and ulcerative keratitis accompanied by scarring resulted. The lesion progressed more quickly in recipients of lymphoid cells specifically immune to HSV and containing cytotoxic T-lymphocyte activity. In such mice, necrosis and ulceration were marked on the sixth day after transfer compared with 9-12 days for those given nonimmune cells. Removal of T-lymphocytes from the immune lymphoid population by treatment with specific antiserum and complement abrogated the adoptive transfer of the stromal reaction. Our results further demonstrate that stromal keratitis represents a host immunopathologic response to HSV infection in which T-lymphocytes are essential participants. Multiple mechanisms of T-cell immunopathology appear to be operating, including a reaction mediated by cytotoxic T-lymphocytes.

Animals↗
PubMed1983

Frequency of cytotoxic T lymphocyte precursors to herpes simplex virus type 1 as determined by limiting dilution analysis.

The conditions for establishing a limiting dilution assay to measure cytotoxic T lymphocyte precursors (CTL-P) against herpes simplex virus type 1 (HSV-1) were determined. Analysis by Poisson statistics demonstrated that the estimated frequency of HSV-1-reactive cells in the spleens of normal mice was less than 1/250,000. In contrast, mice immunized previously with infectious HSV-1 demonstrated a CTL-P frequency between 1/3,500 and 1/15,670. The generation of a maximum cytotoxic T lymphocyte response required that mice be primed in vivo with infectious virus. Immunization with inactivated virus either failed to elicit detectable CTL-P frequencies or gave frequencies markedly less than those induced with infectious virus. To obtain positive cultures, the responder cell population had to be exposed to stimulator splenocytes expressing viral antigens. Normal splenocytes without virus or normal splenocytes with T cell growth factor did not result in significant cytotoxicity. Split culture analysis comparing cytotoxicity against syngeneic and allogeneic virus-infected targets provided evidence for specificity, H-2 restriction, and the T cell nature of the CTL-P. It was determined that precursors were eliminated by treatment with anti-Thy 1, Lyt 2.1, or Lyt 1.1, indicating the CTL-P were Lyt 1(+)2(+) cells. Cytotoxicity was reduced after treatment of the responders with anti-Lyt 2 plus complement, which gave further evidence of the T cell nature of the cytotoxic T lymphocytes. These experiments demonstrated the feasibility of using the limiting dilution approach as a highly sensitive and quantitative means to measure the cell-mediated immune response to HSV-1 antigens.

Animals↗
PubMed1983

Recovery from lethal herpes simplex virus type 1 infection is mediated by cytotoxic T lymphocytes.

The ability of herpes simplex virus (HSV)-specific, cytotoxic T lymphocytes to mediate recovery of mice lethally infected with HSV was examined. Adoptive transfer of splenocytes from mice that had been primed in vivo with HSV and restimulated with HSV in vitro protected lethally infected normal and cyclophosphamide-immunosuppressed mice from death. In contrast, equal numbers of normal splenocytes or immune splenocytes cultured without antigen failed to mediate recovery. Recovery was also transferred by noncultured, primary immune splenocytes, although the protective efficacy of these cells was 10-fold less than when immune splenocytes restimulated in vitro were used. Treatment of the cells with anti-Thy 1 or anti-Lyt 2.1 plus complement before adoptive transfer abrogated recovery. No decrease in protection was seen when in vitro-restimulated splenocytes were treated with anti-Lyt 1.1 plus complement. Splenocytes expression natural killer activity also failed to effect recovery. The administration of hyperimmune anti-HSV antibody to normal, immunocompetent mice resulted in recovery, whereas no significant protection of immunosuppressed mice by anti-HSV was observed. When antibody was given concurrently with cultured immune splenocytes, the percentage of mice that recovered from infection was greater than that seen with either antibody or cultured immune splenocytes alone. These experiments demonstrate that Lyt 2-positive cells with cytotoxic activity generated by in vitro immunization can mediate recovery from lethal HSV infection, whereas, under the conditions chosen, Lyt 1-positive cells were unable to mediate recovery.

Animals↗
PubMed1983

Cellular interactions in the cytotoxic T lymphocyte response to herpes simplex virus antigens: differential antigen activation requirements for the helper T lymphocyte and cytotoxic T lymphocyte precursors.

The role and induction requirements of helper T lymphocyte responses to herpes simplex virus type 1 (HSV-1) was examined. Splenocytes from mice that had been primed in vivo with infectious HSV-1 can be restimulated in vitro with live or partially UV-inactivated HSV-1 to generate high levels of herpes virus-specific cytotoxic T lymphocyte (CTL) activity. By comparison, naive splenocytes or splenocytes taken from mice primed with heat-inactivated HSV-1 failed to generate CTL after in vitro viral stimulation. In addition, infectious HSV-primed splenocytes can be rendered unresponsive to secondary in vitro restimulation by pretreatment with anti-Lyt-1 antiserum plus complement. Spleen cells were taken from mice that had been primed and restimulated in vivo with infectious HSV-1. Two days after the second priming, splenocytes were prepared and irradiated. These cells were capable of assisting in the generation of CTL to varying degrees in all of the above unresponsive populations of cells. The irradiated cells did not produce detectable levels of CTL activity when cultured alone with antigen. Also, if the irradiated splenocytes were treated with anti-Lyt-1 plus complement before their addition to cultures, all restorative activity was ablated. In contrast, irradiated splenocytes from mice that had been primed and restimulated in vivo with either heat-inactivated or UV-inactivated HSV-1 were unable to provide help to naive or helper-depleted cultures. The failure to supply helper activity appears not to involve the preferential activation of suppressor cells, as evidenced by cell mixing experiments and the addition of concentrated, antigen-stimulated spleen cell supernatant fluids to secondary anti-HSV-1 splenocyte cultures. Proliferative assays using interleukin 2- (IL 2) dependent cell lines as a measure of relative helper activity indicated that the inactivated forms of HSV-1 were incapable of effectively enlisting helper activity. These experiments therefore suggest that the observed failure of heat-inactivated or UV-inactivated HSV-1 preparations to induce anti-HSV CTL responses reflects the inability of the HSV-1-specific subset of helper T lymphocytes to recognize these forms of the antigen.

Animals↗
PubMed1983

Enhanced responsiveness of committed macrophage precursors to macrophage-type colony-stimulating factor (CSF-1) induced in vitro by interferons alpha + beta 1.

Contrasting effects of interferon alpha + beta (IFN) on colony-stimulating factor- (CSF) induced macrophage stem cell proliferation resulted when nucleated murine femoral marrow cells were exposed to IFN either before addition of CSF or simultaneously with CSF. Responsiveness to macrophage type L cell CSF was measured by two different assays: by determination of [3H]thymidine (3H-TdR) incorporation and by enumeration of colonies formed in response to CSF. Incubation of nucleated marrow cells concurrently with CSF and IFN resulted in depressed proliferation that was dependent on the dose of IFN. Preincubation of marrow cells with IFN, however, produced a dose-dependent enhancement of CSF responsiveness in comparison to preincubation of marrow cells in the absence of IFN. The enhancing effect of IFN was observed only when preincubated cells were stimulated with suboptimal concentrations of CSF. At optimal CSF concentrations, differences were not observed in either 3H-TdR incorporation or in colony formation. On the basis of data accumulated by both assays for CSF responsiveness, IFN pretreatment was hypothesized not to increase the total population of existing progenitor cells, but to influence the capacity of pre-existing progenitor cells to respond to suboptimal doses of the growth factor.

Animals↗
PubMed1982

Dissection of the proliferative and differentiative signals controlling murine cytotoxic T lymphocyte responses.

Evidence is presented that interleukin 2 (IL-2) is not sufficient to cause the differentiation of primary cytotoxic T lymphocytes (CTL). Sources of IL-2 were compared for their ability to cause proliferation as well as differentiation into CTL. Whereas all factors caused proliferation, only the crude Con A supernatant had cytotoxic T cell differentiation factor (CTDF) activity. Furthermore, factors absorbed with an IL-2-dependent cell line to remove IL-2 still retained CTDF activity. Thus, IL-2 functions to cause clonal expansion of CTL precursors preactivated by antigen or mitogen, but for their differentiation into CTL, an additional factor is required, here called CTDF.

Animals↗
PubMed1982

In vivo induction of anti-herpes simplex virus immune response by type 1 antigens and lipid A incorporated into liposomes.

To establish the requirements for a potential subunit vaccine against herpes simplex virus (HSV), we analyzed the effects on immunogenicity of incorporating detergent-extracted glycoprotein-enriched HSV type 1 (HSV-1) antigens into liposomes alone or with the adjuvant lipid A. Incorporating HSV-1 antigens into liposomes enhanced their immunogenicity for antibody production as detected by radioimmunoassay. Antibody levels to free and liposome-bound antigens were enhanced by administering lipid A as an adjuvant. The maximum immunogenic effect was obtained by incorporating lipid A into liposomes containing the HSV-1 proteins. Such liposomes induced secondary antibody responses higher than those engendered by virus infection. Whereas infectious virus induced cell-mediated immunity detectable by the delayed-type hypersensitivity reactions and cytotoxic T lymphocyte production, none of the liposome preparations induced cell-mediated immunity.

Adjuvants, Immunologic↗
PubMed1982

Role of Ia antigen expression and secretory function of accessory cells in the induction of cytotoxic T lymphocyte responses against herpes simplex virus.

Splenocytes from mice which have been primed in vivo with herpes simplex virus type 1 (HSV-1) can be restimulated in vitro with infectious or UV-inactivated HSV-1 to generate HSV-specific, H-2-restricted cytotoxic T lymphocytes (CTL). HSV-primed splenocytes which have been depleted of adherent cells by sequential incubation on plastic, nylon wool, and Sephadex G-10 are not able to respond with a CTL response when restimulated in vitro. A variety of Ia-positive and Ia-negative (Ia(+) and Ia(-)) cell populations were assessed for their ability to supply accessory cell functions to spleen cells which had been exhaustively depleted of adherent cells, as measured by the restoration of a CTL response to HSV-1. Of these, only those populations which included Ia(+) cells were capable of providing accessory cell function. The ineffective populations were devoid of Ia antigens, except for B lymphocytes, which are Ia(+) and still incapable of serving as accessory cells. Splenic adherent cells and resident peritoneal cells were both proficient at restoring anti-HSV CTL responses, although splenic adherent cells were clearly superior at limiting cell numbers. Neither population was capable of accessory cell activity, however, if it was pretreated with anti-Ia antiserum plus complement or if anti-Ia serum was present during induction. Peritoneal cells lose virtually all of their membrane-associated Ia antigen after a brief period of in vitro culture (24 to 48 h). Cultured peritoneal cells, as well as P388D(1) cells (normally Ia(-)), can be induced to express Ia antigens within 48 h if they are cultured with concanavalin A-stimulated spleen cell supernatants. Ia(+) P388D(1) cells without the extraneous macrophage factor are not able to restore CTL responsiveness to HSV-1 in vitro, whereas Ia(+) cultured macrophages are fully competent accessory cells. When Ia(+) P388D(1) cells were supplemented with the macrophage-derived soluble factor interleukin 1, they displayed a modest, but significant, capacity to restore secondary anti-HSV CTL responses. In addition, glutaraldehyde-fixed, HSV-1-infected Ia(+) peritoneal cells, which could not restore the CTL response alone, were capable of providing accessory cell function if extraneous interleukin 1 was provided. In contrast, Ia(-) cultured peritoneal cells, Ia(-) P388D(1) cells, and various other Ia(-) cell lines were unable to participate in the generation of CTL even in the presence of interleukin 1. The adherent cell population would therefore appear to be making at least two essential contributions to the process of CTL development, namely, the secretion of interleukin 1 and the presentation of antigen in the context of membrane-associated Ia antigen.

Animals↗
PubMed1981

Effect of herpes simplex virus infection on murine antibody-dependent cellular cytotoxicity and natural killer cytotoxicity.

Mice intraperitoneally inoculated with a sublethal dose of herpes simplex virus (HSV) produced immunoglobulin G antibody-dependent cellular cytotoxicity (ADCC) and radioimmunoassay (RIA) antibody as early as 3 days after infection. There was a rise in natural killer cytotoxicity (NKC) to infected and uninfected target cells 1 to 3 days postinfection mediated by nonadherent peritoneal cells (PC) in mice inoculated with HSV, but also with other substances commonly used in tissue culture media. HSV caused the highest and most consistent increase in NKC. PC-NKC, as ADCC, was inhibited by latex and silica, both macrophage inhibitors. PC-ADCC markedly declined 3 to 8 days after HSV inoculation. This was not due to a soluble or cellular suppressor factor, was not reversed by incubation or trypsin treatment of PC, was not associated with a change in PC Fc receptors, adherence, or acridine orange staining characteristics, and could not be induced by inactivated HSV. In vitro inoculation of PC with HSV similarly caused a reduction in the ability of PC to mediate ADCC to HSV-infected target cells. These data demonstrate the complex stimulatory and inhibitory interactions between virus and host defense mechanisms.

Animals↗
PubMed1981

Cell-mediated immunity to herpes simplex virus: recognition of type-specific and type-common surface antigens by cytotoxic T cell populations.

In this communication, we examine the specificity of anti-herpes simplex virus (HSV) cytotoxic T lymphocytes (CTL). Serological studies of the two related HSV serotypes (HSV-1 and HSV-2) have revealed both type-specific and cross-reactive antigenic determinants in the viral envelope and on the surface of infected cells. By analysis of cytotoxicity of CTL, generated in vitro by restimulation of splenocytes from mice primed with one or the other HSV serotype, the recognition of both type-specific and cross-reactive determinants on infected target cells by anti-HSV CTL was detectable. Thus, effector cells generated by priming and restimulating with the same virus recognized both type-specific and cross-reactive determinants on target cells infected with the homologous virus, but only cross-reactive determinants on target cells infected with the heterologous HSV serotype. CTL generated by restimulation with the heterologous virus were capable of recognizing only the cross-reactive determinants on either HSV-1- or HSV-2-infected target cells. These results indicate that two subpopulations of CTL exist in a population of anti-HSV immune spleen cells--those which recognize type-specific determinants and those specific for cross-reactive antigenic determinants present on the surface of HSV infected cells. The type-specific subset of anti-HSV CTL was shown to recognize the gC glycoprotein of HSV-1 infected target cells. In addition to the gC glycoprotein, at least one other type-specific surface antigen was also recognized by anti-HSV CTL in addition to the cross-reactive determinants recognized by anti-HSV CTL.

Antigens, Surface↗
PubMed1981

The role of accessory cells and T cell-growth factor in induction of cytotoxic T lymphocytes against herpes simplex virus antigens.

The roles of accessory cells and T cell-growth factor (TCGF) in the in vivo induction of herpes simplex type 1 (HSV) specific cytotoxic lymphocytes (CTL) were evaluated. Spleen cells from animals infected with HSV 4-6 weeks previously were depleted of adherent cells by passage over Sephadex G10. Unlike intact cells, such depleted spleen cells failed to respond by producing H-2 restricted virus-specific CTL upon culture for 5 days with infectious HSV. The CTL response could be restored either by adding normal genetically compatible peritoneal cells as accessory cells or by the addition of TCGF. To obtain optimum restoration accessory cells needed to be added soon after culture initiation but with TCGF addition, partial restoration was evident when added as late as 72 hr after culture. TCGF also permitted intact spleen cells to respond to heat-inactivated virus. The results are interpreted to indicate that accessory cells are essentially required for the presentation of virus to specific helper cells with such cells responding by the production of TCGF. The results also indicate that certain forms of virus may trigger the response of CTL precursors but not the response of helper cells.

Animals↗
PubMed1981

Cell-mediated immunity to herpes simplex virus: induction of cytotoxic T lymphocyte responses by viral antigens incorporated into liposomes.

The immunogenicity of inactivated herpes simplex virus type 1 (HSV-1) antigens incorporated into liposomes was measured by their ability to induce secondary anti-HSV-1 specific cytotoxic T lymphocyte (CTL) responses in splenocyte cultures from virus-primed mice. Such virus-specific CTL could be induced provided the liposomes contained virus along with plasma membrane antigen of the same H-2 type as that of the virus-primed responser cells. Responses did not occur in cultures stimulated with liposomes containing only viral antigens or with a mixture of liposomes composed respectively of lipid and virus with those composed of lipid and plasma membrane proteins. Moreover F1 responder cells stimulated with liposomes composed of virus and plasma membrane protein of one of the parental haplotypes produced CTL restricted in their cytotoxicity to infected targets of the same haplotype as was used in the liposome. These results show that liposomes can be used to induce anti-HSV-1 CTL with inactivated viral antigens but recognition of both viral and H-2 antigen is required for this process to occur in vitro. The implications of our findings to the preparation of subunit vaccines against HSV-1 are briefly discussed.

Animals↗
PubMed1980

Neutrophils in antiviral immunity: inhibition of virus replication by a mediator produced by bovine neutrophils.

Neutrophils collected from bovine mammary glands were placed in culture with virus-infected cell preparations that had been inactivated by ultraviolet light. Upon culture with infectious bovine rhinotracheitis (IBR) virus-infected Georgia bovine kidney cells, a material was released from the neutrophils that, like interferon, inhibited the replication of vesicular stomatitis virus and IBR virus. Cell-free IBR virus was a less effective inducer of the inhibitor produced by neutrophils. Other herpesviruses (including herpes simplex type 1), as well as equine rhinopneumonitis virus and pseudorabies virus, caused release of the inhibitor, but most other viruses tested, including Newcastle disease virus, a good inducer of type 1 interferon, failed to stimulate the production of inhibitor by polymorphonuclear neutrophils. Preliminary characterization studies revealed differences between the inhibitor produced by the polymorphonuclear neutrophils and type 1 and type II bovine interferons. The possible role of such an inhibitor in the recovery from herpesvirus infection is briefly discussed.

Animals↗
PubMed1980

Cell-mediated immunity against herpes simplex induction of cytotoxic T lymphocytes.

The conditions required for the induction of both primary cytotoxic T lymphocytes (CTL) in vivo and secondary CTL in vitro against herpes simplex virus type 1 (HSV-1)-infected cells were defined. Primary CTL responses occurred only in mice exposed to infectious HSV-1. These responses, which were shown to be mediated by T lymphocytes, peaked at 1 week and had disappeared by 2 weeks after infection. The level of primary cytotoxicity was enhanced by treatment of mice with cyclophosphamide before infection. Secondary in vitro CTL responses were more pronounced and were induced by some forms of inactivated virus as well as by infectious HSV-1. Thus, both ultraviolet light- and glutaraldehyde-inactivated preparations of HSV-1 induced CTL, but heat-inactivated and detergent-extracted antigens failed to do so. The reasons for the differing efficiency of infectious and noninfectious HSV-1 for induction of CTL are discussed.

Animals↗
PubMed1980

Cell-mediated immunity to herpes simplex virus: specificity of cytotoxic T cells.

This communication deals with the question of which of the viral antigens constitutes the targets for cytotoxic T lymphocytes (CTL) generated against herpes simplex virus type 1 (HSV-1). The approach used was, first, to compare cytotoxicity of CTL against target cells infected with virus in the presence of tunicamycin and 2-deoxy-D-glucose, which are known to inhibit glycoprotein synthesis, and second, to compare cytotoxicity of CTL against target cells infected with wild-type HSV-1 with that against target cells infected with a temperature-sensitive mutant of HSV-1 which, at the nonpermissive temperature, exhibits diminished glycoprotein synthesis. The results show that glycoprotein expression is required for the demonstration of cytotoxic activity of CTL. The level of cytotoxicity against the temperature-sensitive HSV-1 target at the nonpermissive temperature was reduced and correlated with the level of expression of the major envelope glycoprotein region (VP123; molecular weight = 123,000) at the target cell surface as measured serologically by antibody binding studies. The results were interpreted to indicate that HSV-1-induced glycoproteins are the target antigens for anti-HSV CTL and that the principal viral antigens recognized by the CTL may be glycoproteins of the VP123 region.

Animals↗