Search PubMed⌕ Search

Biomedical subjects

B T Rouse

Publications and source records attributed to B T Rouse.

At least 163 records · Page 9Linked to original sources
PubMed1986

Estimation of the B lymphocyte precursor frequencies to herpes simplex type 1 glycoproteins by a limiting dilution assay.

The precursor frequency of B lymphocytes from Balb/c mice producing HSV-1 glycoprotein B (gB), glycoprotein C (gC), and glycoprotein D (gD) antibody was determined by limiting dilution analysis under conditions to detect antibody from the clonal progeny of a single B cell precursor. In spleens of naive mice the average gC frequency was 1/48,917 +/- 5,550, while gD was 1/73,330 +/- 15,898, and gB frequency was in excess of 1/100,000. Immunization with live HSV-1 (KOS) increased the B cell frequencies of all three glycoproteins to approximately 1:3,000; however, the serum gB antibody ELISA titer was fivefold higher than gC or gD.

Animals↗
PubMed1986

Virus-induced immunosuppression.

Infection with a variety of viruses results in the suppression of the host's immune system. Several mechanisms thought to be responsible for this effect are discussed: infection and alteration of lymphocytes and macrophages, production of soluble suppressor factors, and the induction of suppressor cells. The clinical significance of virus-induced immunosuppression is also discussed.

Animals↗
PubMed1986

Regulation of herpes simplex virus-specific cell-mediated immunity by a specific suppressor factor.

Our study was designed to investigate the nature of an antigen-specific suppressor factor generated by antigen-stimulated herpes simplex virus (HSV)-immune splenocytes. Factor SF-200, a 90,000- to 100,000-dalton fraction obtained after Sephacryl gel filtration, suppressed the generation of HSV-specific cytotoxic T-lymphocyte and lymphoproliferative responses. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis of SF-200 indicated that it contained an I-J+, anti-idiotypic protein. It was possible to adsorb the suppressor activity of SF-200 to an anti-I-J immunoaffinity column. The suppressor activity could be eluted from the immunoaffinity column with a low-pH buffer. The acid-eluted material was determined to be both I-J+ and reactive with anti-HSV antiserum by Western blot analysis. Both SF-200 and the I-J+ suppressor activity suppressed only HSV-specific cell-mediated immunity responses. However, it was possible to generate nonspecific suppressor activity by incubating the I-J+ suppressor factor with Lyt 1+ splenocytes from HSV-immune mice. The implication of these results with respect to the model for a suppressor cell circuit regulating HSV-specific cell-mediated immunity responses is discussed.

Animals↗
PubMed1986

Targeting of drug loaded immunoliposomes to herpes simplex virus infected corneal cells: an effective means of inhibiting virus replication in vitro.

The goal of our studies was to develop liposomes containing antiviral drugs and targeted with antiviral antibody (immunoliposomes) that would be effective at inhibiting replication of herpes simplex virus (HSV) in vitro. To achieve this, a monoclonal antibody to glycoprotein D of HSV was derivatized with palmitic acid and was incorporated into the lamellae of dehydration-rehydration vesicles. The gD containing immunoliposomes were shown to bind specifically to HSV-infected rabbit corneal cells in vitro, whereas control immunoliposomes prepared with a monoclonal antibody of the same class as the anti-gD failed to preferentially bind to virus-infected cells. The gD immunoliposome binding was inhibitable by pretreatment with rabbit anti-HSV serum but not by aggregated normal serum. Thus liposome binding was judged to represent an antigen-antibody reaction not binding to Fc receptors expressed by cells infected with HSV. Immunoliposomes loaded with iododeoxyuridine (IUDR) leaked drug rapidly at 37 degrees C, whereas acyclovir (ACV)-loaded liposomes still contained 48% of drug after 24 hr at 37 degrees C. The ACV-liposomes retained 44% of drug after 14 days at 4 degrees C. The ability of immunoliposomes to inhibit virus replication was compared with that of untargeted and empty liposomes by means of virus yield assays in vitro, Immunoliposomes loaded with either IUDR or ACV inhibited virus replication, although ACV-containing immunoliposomes were the most efficacious. The implications of our in vitro results for the development of immunoliposomes suitable for the treatment of ocular herpes infection are briefly discussed.

Acyclovir↗
PubMed1986

Production and characterization of an anti-idiotypic antibody specific for a monoclonal antibody to glycoprotein D of herpes simplex virus.

A monoclonal antibody specific for glycoprotein D (gD) of herpes simplex virus type 1 (HSV-1) was used to prepare an anti-idiotypic antibody in rabbits. After removal of antibody reactivity to constant region determinants by absorption with polyclonal mouse immunoglobulins and a monoclonal antibody of the same subclass as the anti-gD monoclonal, the anti-idiotypic (anti-id) antibody reacted specifically with anti-gD. Using an ELISA inhibition assay with immunoaffinity-purified gD, the anti-id D reagent inhibited the binding of anti-gD to gD, suggesting that anti-id D mimics an epitope of gD by binding the antigen-combining site of anti-gD. Immunization of mice with anti-id D could prime splenocytes in vivo to proliferate in response to HSV antigen stimulation in vitro. The possibility that anti-id D could act similarly to gD and stimulate an immune response to HSV when administered in vivo is discussed.

Animals↗
PubMed1985

Interferon produced endogenously in response to CSF-1 augments the functional differentiation of progeny macrophages.

The role of endogenously produced interferon alpha/beta in the functional maturation of newly derived mononuclear phagocytes was investigated. Addition of highly specific anti-interferon alpha + beta antiserum to murine marrow cultures stimulated with colony-stimulating factor-1 (macrophage growth factor) markedly suppressed the capacity of resulting progeny mononuclear phagocytes to ingest opsonized sheep erythrocytes (EAIgG). This impairment was corrected either by direct addition of interferon alpha + beta at a concentration in excess of that neutralized by the antiserum or by the addition of lesser amounts of interferon (33 U/ml) following removal of the anti-interferon from the cultures. Conditioned media from control colony-stimulating factor-stimulated cultures similarly reversed the impairment of maturation resulting from 5 days of growth in the presence of anti-interferon. This enhancement of EAIgG ingestion reflected upon the interferon activity in the conditioned media and was neutralized by anti-interferon. Lastly, the endogenous interferon was found to enhance EAIgG ingestion by a majority of the mononuclear phagocyte progeny and not by a limited subpopulation.

Animals↗
PubMed1985

Herpes simplex virus-specific lymphoproliferation: an analysis of the involvement of lymphocyte subsets.

Herpes simplex virus (HSV)-immune murine splenocytes incorporated significant levels of tritiated thymidine when incubated with UV-inactivated, heat-inactivated, and active preparations of HSV. Normal splenocytes incubated with the HSV preparations did not exhibit such proliferation. Maximum incorporation by the immune splenocytes occurred on the fifth day of culture and was mediated by Thy-1+, Lyt-1+, and Lyt-2+ cells. Attempts to correlate lymphoproliferation with other HSV-specific cellular immune responses demonstrated the complexity of this response. While T cells mediating delayed type hypersensitivity responses and cytotoxic T lymphocytes were involved in the lymphoproliferative response, neither could be considered as being exclusively associated with lymphoproliferation. Instead, lymphoproliferation appeared to be indicative of HSV-specific Lyt-1+ helper cells. Evidence was also presented that suppressor cells appeared to be involved in the regulation of the lymphoproliferative response.

Animals↗
PubMed1985

Regulation of interleukin 1 production by alpha and beta interferons: evidence for both direct and indirect enhancement.

In this paper, the influence of interferons alpha + beta (IFN alpha + beta) on the levels of interleukin-1 (IL-1) produced by murine peritoneal exudate macrophages following subsequent stimulation with lipopolysaccharide (LPS), colony-stimulating factor-1 (CSF-1), or both was investigated. The results indicate that preincubation with IFN alpha + beta enhances IL-1 production in response to CSF-1 as well as to LPS. Moreover, prior exposure to IFN alpha + beta enhanced the apparently synergistic production of IL-1 in response to the two stimuli combined. Lastly, IFN alpha + beta in dosages exceeding 1,000 u/ml appeared to directly stimulate IL-1 production by exudate macrophages. Direct and augmented stimulation of IL-1 production by IFN alpha + beta could be neutralized by addition of highly specific anti-IFN alpha + beta antiserum. The significance of these findings in terms of the reported inflammatory and pyrogenic effects of IFN is discussed.

Animals↗
PubMed1985

Production of soluble suppressor factors by herpes simplex virus-stimulated splenocytes from herpes simplex virus-immune mice.

Indirect evidence indicates that herpes simplex virus (HSV)-specific cytotoxic-T-lymphocyte induction is regulated by suppressor cells. To search for such suppressor effects, supernatant fluids from splenocyte cultures from normal and HSV-immune mice cultured either with or without viral stimulation were tested for their ability to inhibit HSV-specific cytotoxic-T-lymphocyte induction. Only the supernatant fluid from the HSV-stimulated, HSV-immune cultures contained a suppressor activity (HSV-SF). HSV-SF was produced by nylon-wool-purified Thy 1+ cells. HSV-SF was detectable after 3 days of culture and would only suppress cytotoxic-T-lymphocyte induction if HSV-SF was added within 24 h of initiation of the test cultures. HSV-SF was neither dialyzable nor heat stable. Molecular sieve chromatography of HSV-SF yielded multiple peaks of suppressor activity. Although most of these peaks exhibited nonspecific suppressor activity, the suppression mediated by the 90,000 to 150,000-molecular-weight fractions was antigen specific and genetically restricted. These results provide direct evidence for the regulation of HSV-cytotoxic-T-lymphocyte induction by a novel suppressor factor.

Animals↗
PubMed1985

Regulation of herpes simplex virus-specific lymphoproliferation by suppressor cells.

We investigated the regulation of the herpes simplex virus (HSV)-specific lymphoproliferative response (LPR) by suppressor cells. The chief cell types in HSV-immune splenocytes proliferating in response to the antigen were Lyt 1+ and Lyt 2+ T cells, which accounted for approximately 60 and 40% of the response, respectively. Because the total responsiveness of splenocytes was enhanced after depletion of Lyt 2+ cells, the LPR was assumed to be subject to regulation by an Lyt 2+ suppressor cell. This was shown to be the case with an experimental design in which suppressor cell activity was induced in one culture, the cells were irradiated, and the effects on LPR were measured in a test antigen-stimulated culture. The cell responsible for suppression was shown to be Lyt 2+ IJ+, and the actual suppressor effect was not antigen specific. Cellular requirements for the generation of suppression were also investigated. The three distinct cell types that appeared to be required were Lyt 2+ and Lyt 1+ T cells and an IJ+ antigen-presenting cell. Of the three cell types, only the Lyt 2+ cell needed to be from HSV-immune animals. The implications of our model system for the better understanding of the role of immunity in herpesvirus pathogenesis are discussed.

Animals↗
PubMed1985

Frequency of herpes simplex virus-specific helper T lymphocyte precursors in the lymph node cells of infected mice.

We have established a limited dilution assay to estimate the frequency of herpes simplex virus type 1 (HSV)-specific, interleukin 2 (IL 2)-producing helper T lymphocyte precursors (HTL-P). The estimated frequency of such cells in suspensions of local lymph node (LN) cells 5 days after in vivo virus infection was 1:2470 to 1:5800. Frequencies of HTL-P in cells from uninfected mice were below levels of detection of our system and were judged to be below 1:100,000. Removal of Lyt-2+ cells from responder LN cells before culture increases HTL-P frequency twofold to threefold, indicating the likely operation of some form of suppression in unseparated cultures. The demonstration of HSV-specific HTL-P required that cells from virus-primed mice be reexposed in vitro to viral antigen. In addition, clones expanded during a 9-day culture period failed to generate IL 2 unless reexposed to specific viral antigen or cross-reactivate HSV-2. Thus, HSV-specific HTL-P were strictly antigen dependent. No evidence was obtained for antigen-independent subpopulations of HTL-P as occurs with viral-specific cytotoxic T lymphocyte precursors. The clonal progeny of HTL-P were of the Thy-1+ Lyt+2- phenotype. Priming in vivo for the subsequent in vitro detection of HTL-P required that mice be exposed to infectious virus. Thus neither UV-inactivated nor heat-inactivated nor extracted viral glycoproteins could prime for HTL-P detection. The relevance of these findings for the future use of subunit vaccines against HSV is briefly discussed.

Animals↗
PubMed1985

Augmentation of immunity to herpes simplex virus by in vivo administration of interleukin 2.

The immune mechanisms responsible for recovery from herpesvirus infections are multiple and include a principle role for aspects of T cell immunity. Our investigations add further support for this notion. We show that the ability of immune lymphocytes from animals infected i.p. 6 wk previously with herpes simplex virus type one (HSV-1) clear virus more effectively when interleukin 2 (IL 2) is injected into recipients of the adoptive transfers. Mice were treated on two consecutive days with cyclophosphamide and infected in the pinnae with 4 X 10(6) plaque-forming units of HSV-1. Three hours post-infection lymphocyte populations were injected i.v., and after a further 3 days the pinnae were removed, homogenized, and the content of infectious virus assayed. Purified IL 2 obtained from EL-4 cells either was given i.v. 2 hr before and 24 and 48 hr after cell injection or was given subcutaneously 2 hr before and 3, 24, and 48 hr after cell injection. The latter three injections were given i.p. and suspended in 15% gelatin. The immune lymphocyte cell populations were splenocytes and were either injected immediately after preparation of cell suspensions or after 5 days in vitro secondary stimulation with HSV-1. This latter cell population showed greater viral clearance activity, a function shown previously to be a property of Lyt-2+ cells. The clearance activity of cells was markedly enhanced in animals given IL 2 but only with a regimen that included injections in gelatin, a procedure that enhances in vivo circulation time of IL 2. The cell involved in clearance was a T cell and principally the Lyt-2+ subset. Treatment of recipient mice with anti-asialo GM-1 did not affect the clearance efficiency, indicating that NK cells were not responsible for the observed effect. Our experiments indicate that IL 2 may provide an important regulator of immune function in vivo and may warrant its investigation as a therapeutic agent to enhance antiviral immunity in certain circumstances.

Adjuvants, Immunologic↗
PubMed1985

Endogenous regulation of macrophage proliferation and differentiation by E prostaglandins and interferon alpha/beta.

Our results indicate that there are at least two levels of interaction involving M-CSF stimulation of committed stem cells and mature mononuclear cells. Mature cells residing in hematopoietic tissues produce both PGE and IFN alpha/beta upon recognition of CSF in their environment. In vitro, both of these mediators act to suppress CSF-induced colony formation in a negative feedback manner. However, additional evidence indicates that the endogenous IFN acts also to enhance functional maturation of the monocyte progeny. Cultures deprived of IFN during clonal expansion displayed severely depressed capacities to produce IL-1, to resist lytic HSV infection, and to phagocytose opsonized erythrocytes via Fc receptors. Thus, it appears that the result of M-CSF stimulation of a mature cell is the production of a differentiation signal (IFN alpha/beta), which in turn affects an immature cell of the same lineage that responds to the CSF-1 growth stimulus. Whether the Ia+ subpopulation of M-CSF responsive progenitor cells represents precursors for a separate cell lineage or a subclass of macrophages remains to be determined. Similarly, the regulation of these cells by PGE and IFN remains to be explored. However, within the parameters of our investigation, neither these cells nor their specific regulator, acidic isoferritin, were of consequence.

Cell Differentiation↗
PubMed1984

Endogenous regulation of macrophage proliferative expansion by colony-stimulating factor-induced interferon.

Stimulation of cultures of murine bone-marrow cells with specific macrophage growth factor (colony-stimulating factor I) resulted in the production of type I interferon. Neutralization of this endogenous interferon by antiserum directed against interferons alpha and beta resulted in a significant enhancement of mononuclear phagocyte proliferation from committed marrow precursors. The effect of the antiserum was lost in cultures depleted of adherent cells, an indication that an adherent regulatory cell (or cells) in the marrow limits mononuclear phagocyte proliferation by producing antiproliferative interferon in response to high levels of specific growth factor.

Animals↗
PubMed1984

Role of T-lymphocyte subsets in recovery from herpes simplex virus infection.

Our investigations probed the nature of different T-lymphocyte subsets effecting clearance of herpes simplex virus after infection of the pinna. Cell populations from animals recently infected subcutaneously or intraperitoneally (acute population) or from animals infected 6 weeks previously (primed population) or the latter cells reimmunized in vitro with virus (memory population) were studied. Viral clearance was a function of the Lyt 1+2- subset in the acute population, but with the memory population both Lyt 1+ and Lyt 2+ cells affected clearance. In primed populations, viral clearance was effected only by the Lyt 2+ subset. The ability of the various cell populations to adoptively transfer delayed-type hypersensitivity was also studied. Only acute population cells from animals infected subcutaneously and memory population cells transferred delayed-type hypersensitivity. In both cases, the cell subtype was Lyt 1+2-. Our results demonstrated that the delayed-type hypersensitivity response does not always correlate with immunity to herpes simplex virus. Multiple subsets of T cells participate in viral clearance, and their respective importances vary according to the stage of the virus-host interaction.

Animals↗
PubMed1984

Frequency of herpes simplex virus-specific cytotoxic T lymphocyte precursors in lymph node cells of infected mice.

The limiting dilution technique was used to estimate the frequency of cytotoxic T lymphocyte precursors (CTL-P) specific to herpes simplex virus (HSV)-infected syngeneic cells in draining lymph nodes from CBA mice infected 5 days previously with HSV. Significant numbers of virus specific CTL-P were found with infected mice whereas such cells were usually undetectable in uninfected mice. On the basis of MHC restriction and Lyt phenotype, the cells were identified as T lymphocytes. Two subsets of HSV-specific CTL-P could be defined; an infrequent one (1/12,500-1/30,000) which grew in the presence of growth promoting factors present in concanavalin A-conditioned medium, and a frequent population (f = 1/1200-1/4750) which in addition required exposure to antigen. In some experiments, the frequent subset of HSV-specific CTL-P was only detectable at low cell number inputs. Similar results in other systems have been explained by active suppression.

Animals↗