Biomedical subjects

R M Gorczynski

Publications and source records attributed to R M Gorczynski.

At least 55 records · Page 3Linked to original sources

PubMed1998

Donor-specific stimulation of peripheral blood mononuclear cells from recipients of orthotopic liver transplants is associated, in the absence of rejection, with type-2 cytokine production.

In this study, we examined the cytokine production by human peripheral blood mononuclear cells (PBM) from recipients of orthotopic liver transplants which had been stimulated by donor-specific alloantigen. Levels of interleukin (IL)-2, IL-4, interferon (IFN)-gamma, IL-10 and transforming growth factor (TGF)-beta produced in vitro from PBM of 15 transplant recipients at 5-7 months post transplantation were analysed after donor-specific, third-party, or non-specific stimulation. Mononuclear cell proliferation in response to stimulation and cytokine mRNA from the cell cultures were assayed. Donor-specific antigen was obtained from donor spleen cells which had been obtained and frozen in liquid nitrogen at the time of organ retrieval. Third-party restimulation used equivalent numbers of spleen cells pooled from the other 14 organ donors. Cytokine production was correlated with the clinical condition of the patient, including biopsy results when available, and biochemical data. The data show a highly significant correlation between the donor-specific- and third-party- stimulated IL-4 and IL-10 production from recipient PBM with stable liver graft function as assessed by histopathology and/or biochemistry. This correlation was independent of level of immunosuppression. These data strongly support a role for IL-4 and/or IL-10 in the induction and/or maintenance of tolerance to human liver allografts. Measurement of the levels of these cytokines from recipient PBM after donor-specific antigen stimulation in vitro may be a useful test for monitoring for acute allograft rejection.

Adjuvants, Immunologic↗

PubMed1998

TCR diversity in gammadeltaTCR+ hybridomas derived from mice given portal vein donor-specific pre-immunization and skin allografts.

Portal venous (p.v.) immunization with multiple minor histoincompatible cells leads to antigen-specific increased skin allograft survival. GammadeltaTCR+ hybridoma cells, prepared from mesenteric lymphocytes of p.v. immunized animals, can adoptively transfer this increased graft survival to naive animals. We have analyzed VgammaVdelta gene usage, and TCR gamma-chain junctional diversity in gammadeltaTCR+ hybridomas from mice immunized with different antigen combinations by p.v. or conventional lateral tail vein (i.v.) immunization. Following p.v. immunization two independent sets of hybridoma cells were derived, one expressing a common gamma-chain junctional sequence which was also found in > 85% of the hybridomas derived following i.v. immunization, while the other set showed remarkable gamma-chain junctional sequence diversity. The diversity seen in these latter hybridomas was associated with the antigen specificity of the hybridoma cells. Cells expressing these 'unique' TCR junctional sequences were stimulated to produce cytokines both by hsp and by minor-histocompatibility-specific irradiated peritoneal cells. Cells expressing TCR with a common gamma-chain junctional sequence were stimulated to cytokine production by MHC-matched but minor-histocompatibility mismatched (as well as matched) peritoneal cells, but not by hsp. We suggest that p.v. immunization results in stimulation of both antigen-specific and non-specific regulatory gammadeltaTCR+ cells, which can be distinguished by gamma-chain TCR sequence diversity.

Adoptive Transfer↗

PubMed1998

Analysis of cytokine production and V beta T-cell receptor subsets in irradiated recipients receiving portal or peripheral venous reconstitution with allogeneic bone marrow cells, with or without additional anti-cytokine monoclonal antibodies.

Irradiated (800 rads) AKR mice received intravenous (i.v.) reconstitution with a mixture of B10.BR T-depleted bone marrow cells and spleen cells. Only in groups of mice treated additionally with i.v. cyclophosphamide (Cy; 150 mg/kg), 24 hr before transplantation, was long-term (> 60% at 50 days) survival seen. In mice receiving only irradiation all animals died by 30 days post-transplantation. Histological changes consistent with graft-versus-host disease (GVHD) were seen in the liver of reconstituted mice at 30 days, along with an organ-specific increase in V beta 3 T-cell receptor-positive (TCR+) cells. No such increase in V beta 3 TCR+ cells was seen in the spleen from the same mice. These data are consistent with a tissue antigen-driven expansion of V beta 3 TCR+ cells associated with GVHD in the liver in this model. When we analysed cytokine production in vitro from CD3+ cells restimulated with 'host' (AKR) antigen-presenting cells (APC), we found a transition in cytokine production from preferential synthesis of type-1 cytokines [interleukin-2 (IL-2) and interferon-gamma (IFN-gamma)] at early times (day 15) post-reconstitution to increased production of type-2 cytokines [IL-4, transforming growth factor-beta (TGF-beta) and IL-10] at later times (day 30) post-reconstitution in Cy-treated recipients. Animals not receiving Cy did not show this 'switch' in cytokine production at later time points. We have observed a similar polarization in cytokine production, along with increased graft survival, in recipients of vascularized and non-vascularized allografts after portal venous (p.v.), but not i.v., pretransplant donor-specific immunization. We next studied AKR mice receiving 800 rads and subsequently reconstituted with B10.BR stem cells via the p.v. route. Again these mice showed prolonged survival (> 50% at 50 days), with polarization to IL-4, IL-10 and TGF-beta on restimulation of CD3+ cells in vitro at 30 days post-transplant and increased V beta 3 TCR+ cells in the liver. Infusion of anti-IL-12 monoclonal antibodies into irradiated mice receiving i.v. cell reconstitution produced a similar pattern of changes to those seen after p.v. reconstitution, while a combination of anti-IL-10 and anti-TGF-beta monoclonal antibodies reversed the changes seen after p.v. reconstitution. These data are consistent with an important role for differential cytokine production in the regulation of GVHD following allogeneic bone marrow transplantation.

Animals↗

PubMed1998

Pretreatment of donor stimulator cells by 16,16 dimethyl prostaglandin E2 influences the recipient immune response.

BACKGROUND: Immunosuppressive strategies have largely ignored donor-derived stimulatory cells as a target. This study examined whether lipopolysaccharide (LPS) or 16,16 dimethyl prostaglandin E2 (dmPGE2) pretreatment of stimulator cells from B10.BR mice influences effector function of responder T lymphocytes from C3H/HeJ mice in vitro or in vivo. METHODS: B10.BR spleen cells were incubated in vitro in the presence or absence of dmPGE2 or or LPS before the cells were used as stimulators in a mixed lymphocyte culture (MLC) with T cells from C3H/HeJ mice. In parallel studies, B10.BR mice were treated in vivo with dmPGE2 or LPS; spleen cells from these animals were used as stimulators in an MLC and skin was harvested for skin grafts. Cells from untreated or pretreated mice were examined for expression of intercellular adhesion molecule-1 (ICAM-1), B7-1, and B7-2 by fluorescence-activated cell sorter analysis. ICAM-1 mRNA transcripts were determined by reverse transcriptase-polymerase chain reaction. RESULTS: Stimulation of B10.BR-derived spleen cells with LPS before their use as stimulator cells in a MLC resulted in an increase in responder T-cell proliferation compared with use of unstimulated spleen cells (P < 0.05). In contrast, pretreatment of stimulator spleen cells with dmPGE2 resulted in dose-dependent inhibition of the responder T-cell proliferation, with maximum effect seen using a concentration of dmPGE2 of 10(-5) mol/L. The decreased expression of interleukin-1, tumor necrosis factor, leukotriene B4 procoagulant activity, and ICAM-1 by the dmPGE2 pretreated spleen cells correlated with their inefficient in stimulating T-cell proliferation. Spleen cells harvested from B10.BR mice previously injected with dmPGE2 similarly were inefficient as stimulator cells. Skin graft survival was delayed, but not prevented, by in vivo pretreatment of donor mice with dmPGE2. CONCLUSIONS: These data demonstrate the effect of immunomodulation of allogeneic stimulator spleen cells on subsequent responder T-lymphocyte function and allograft survival.

16,16-Dimethylprostaglandin E2↗

PubMed1997

Cloning and characterization of the murine homologue of the rat/human MRC OX-2 gene.

A 350 bp amplicon, obtained by PCR-select subtractive hybridization from RNA derived from mesenteric lymph nodes (MLN) cells from mice pre-immunized with allogeneic lymphocytes 36hrs prior to receiving donor-specific skin grafts, and showing > 98% homology with a published sequence for the rat MRC OX-2 gene, was used as a hybridization probe to screen a cDNA library constructed from adult mouse MLN treated in the same fashion. Several clones were identified which, on DNA sequence analysis, predicted a 218 amino acid protein showing significant homology with the rat and human MRC OX-2 gene product.

Adoptive Transfer↗

PubMed1997

Specificity for in vivo graft prolongation in gamma delta T cell receptor+ hybridomas derived from mice given portal vein donor-specific preimmunization and skin allografts.

gamma delta TCR+ hybridoma cells prepared from mesenteric lymph node cells of animals receiving donor-specific immunization via the portal vein can adoptively transfer this increased graft survival to naive animals. Analysis of TCR gamma-chain junctional sequence diversity suggested that some 40 to 50% of the hybridomas expressed gamma-chain junctional sequence diversity and were stimulated to produce cytokines both by heat shock proteins and by minor histocompatibility Ag-specific irradiated peritoneal cells. The remaining gamma delta TCR+ hybridoma cells expressed TCR with a common gamma-chain junctional sequence and were stimulated to cytokine production by MHC-matched, but minor histocompatibility Ag-mismatched (as well as matched), peritoneal cells, but not by heat shock proteins. We have compared the effectiveness of representative hybridomas expressing unique gamma-chain junctional sequences or common gamma-chain junctional sequences for prolongation of donor-specific or third-party (MHC-matched or MHC-mismatched) skin grafts. Our data show a good correlation between the specificity for stimulation for cytokine production in vitro and efficacy in graft prolongation assays in vivo. Hybridoma cells expressing unique gamma-chain junctional sequences that showed Ag-specific stimulation of cytokine production in vitro and skin graft survival in vivo augmented survival of third-party skin grafts if simultaneously transplanted with both Ag-specific and third-party skin grafts. Graft prolongation in vivo using cells from either population of gamma delta TCR+ hybridomas was decreased by infusion of anti-IL-10 mAb and abolished when both anti-IL-10 and anti-TGF-beta Abs were used together.

Adoptive Transfer↗

PubMed1997

Conditioned immunosuppression of lipopolysaccharide-induced antibody response of orally immunized mice.

BALB/C mice previously conditioned in a paired regimen in which cycophosphamide was given in association with a novel taste in the drinking water (saccharin, SAC) subsequently received LPS (1mg/mouse) by oral gavage. Subgroups of mice were reexposed to SAC or water respectively, and anti-LPS antibody production measured 6 days later using an ELISA/ELISPOT assay. Our data show that the immune response to this T-independent antigen, administered orally, is amenable to conditioned immunosuppression in this model system.

Administration, Oral↗

PubMed1997

The effect of immunomodulation of stimulator antigen presenting cells on subsequent responder T-cell function.

In this study, we examined whether lipopolysaccharide (LPS)-, interferon-gamma (IFN-gamma) or 16,16-dimethyl prostaglandin E2 (dmPGE2)-pretreatment of stimulator spleen cells from C57BL6 (B6) mice affects effector function of responder T-lymphocyte from C3H/HeJ mice. Stimulation of B6-derived splenic mononuclear cells (SMNCs) with LPS (10 micrograms/ml) prior to their utilization as stimulator cells in a mixed lymphocyte culture (MLC) resulted in an increase in responder T-lymphocyte proliferation compared to utilization of unstimulated SMNC (P < 0.05). IFN-gamma demonstrated similar effects in a dose dependent fashion with maximal stimulatory effect seen at 1000 U/ml. In contrast, pretreatment of stimulator SMNC with dmPGE2 resulted in dose-dependent inhibition of the responder T-lymphocyte proliferation with maximum inhibitory effect seen using a concentration of dmPGE2 of 10(-5) M. The presence of indomethacin in the MLC did not reverse this effect. These data demonstrate the effect of immunomodulation of stimulator spleen cells on subsequent T-lymphocyte function.

16,16-Dimethylprostaglandin E2↗

PubMed1997

An antibody specific for interleukin-6 reverses age-associated changes in spontaneous and induced cytokine production in mice.

A number of quantitative and qualitative changes in the pattern of cytokine production have been reported to accompany the process of ageing in laboratory animals and in human populations, including an increase in serum levels of interleukin-1 (IL-1) and IL-6, as well as increased concanavalin A (ConA)-stimulated production of IL-4, IL-10 and transforming growth factor-beta (TGF-beta), and decreased production of IL-2 from cultured spleen cells. Increased IL-1 and IL-6 production is a feature of splenic adherent cells and peritoneal exudate cells taken from aged mice and stimulated with lipopolysaccharide in vitro. We have asked whether the altered production of lymphocyte-derived cytokines (IL-4, IL-2, TGF-beta) is itself a function of a primary alteration in IL-1/IL-6 production (from macrophage/monocytes) by infusing monoclonal antibodies to these cytokines prior to harvesting cells from aged mice and stimulating the cells in vitro. Anti-IL-6, but not anti-IL-1, reversed the age-associated alteration in lymphocyte cytokine production. The general pattern of cytokine production in aged mice is of a type-2 cytokine type, and thus these data are consistent with the idea that increased production of IL-6 in aged animals is causally implicated in this age-associated polarization to type-2 cytokine production.

Aging↗

PubMed1996

Interleukin-13, in combination with anti-interleukin-12, increases graft prolongation after portal venous immunization with cultured allogeneic bone marrow-derived dendritic cells.

Portal venous (pv) transfusion before transplant with large numbers (100 x 10(6)) of irradiated multiple minor histoincompatible spleen cells (B10.Br) augments allogeneic skin graft survival in C3H mice. We have shown in earlier studies that this is correlated with preferential activation for production of type 2 cytokines (interleukin [IL]-4 and IL-10) and decreased production of type 1 cytokines (IL-2 and interferon [IFN] gamma). We have also shown that recombinant (r)IL-12, in association with anti-IL-10 monoclonal antibody, can reverse in vivo the graft prolongation afforded by pv immunization and the altered cytokine production that follows. Adoptive transfer of inhibition of graft rejection is possible at early times after pv immunization, using plastic adherent cells obtained from the liver of treated mice. We show below that within 4 days of pv immunization, dendritic cells (NLDC-145+) isolated from the thymus, mesenteric lymph node (MLN), and.

Adoptive Transfer↗

PubMed1996

Multiple levels of regulation for self-tolerance in beef insulin transgenic mice.

To characterize the mechanism(s) of tolerance toward soluble self-antigens (Ags), beef insulin (BI) transgenic (Tg) mice were generated in which the transgene was expressed in pancreatic beta-cells. Our previous data showed that: (i) Ag-specific tolerance can be induced and/or maintained in peripheral T cells in thymectomized BI-Tg mice and (ii) CD4+ Th2 regulatory T cells are involved in maintaining peripheral tolerance (by anti-BI antibody response). In this paper, we have further characterized the relationship of low levels of BI expression (10(-10)-10(-11) M) in Th1/Th2 activation. In addition, we have explored intrathymic events associated with tolerance to self-Ags not expressed in the thymus and/or to circulating self-Ags. Limiting dilution analysis showed that there was a significantly higher frequency of BI-specific Th2 cells in Tg mice with a corresponding higher frequency of Th1 cells in non-Tg mice. While there was no transgene expression in the thymus (by RT-PCR), independent studies showed that BI can be processed and presented in the Tg thymus, which correlated with the Ag-specific hyporesponsiveness of mature thymocyes detected in vitro. High-dose rIL-2 (150 U/ml) was able to restore in vitro peripheral T cell response of Tg mice to levels comparable to those of the non-Tg control. Collectively, our data suggest that: (i) there is a differential activation of BI-specific Th1/Th2 cells in vivo in the presence of low Ag concentration; (ii) the thymus may play a role in self-tolerance to Ags whose expression in adults is restricted to the periphery; and (iii) multiple levels of regulation such as thymic selection, peripheral anergy, and active suppression may be involved in tolerance to BI in BI-Tg mice.

Animals↗

PubMed1996

Neuroimmune mechanisms in health and disease: 2. Disease.

In the second part of their article on the emerging field of neuroimmunology, the authors present an overview of the role of neuroimmune mechanisms in defence against infectious diseases and in immune disorders. During acute febrile illness, immune-derived cytokines initiate an acute phase response, which is characterized by fever, inactivity, fatigue, anorexia and catabolism. Profound neuroendocrine and metabolic changes take place: acute phase proteins are produced in the liver, bone marrow function and the metabolic activity of leukocytes are greatly increased, and specific immune reactivity is suppressed. Defects in regulatory processes, which are fundamental to immune disorders and inflammatory diseases, may lie in the immune system, the neuro endocrine system or both. Defects in the hypothalamus-pituitary-adrenal axis have been observed in autoimmune and rheumatic diseases, chronic inflammatory disease, chronic fatigue syndrome and fibromyalgia. Prolactin levels are often elevated in patients with systemic lupus erythematosus and other autoimmune diseases, whereas the bioactivity of prolactin is decreased in patients with rheumatoid arthritis. Levels of sex hormones and thyroid hormone are decreased during severe inflammatory disease. Defective neural regulation of inflammation likely plays a pathogenic role in allergy and asthma, in the symmetrical form of rheumatoid arthritis and in gastrointestinal inflammatory disease. A better understanding of neuroimmunoregulation holds the promise of new approaches to the treatment of immune and inflammatory diseases with the use of hormones, neurotransmitters, neuropeptides and drugs that modulate these newly recognized immune regulators.

Acquired Immunodeficiency Syndrome↗

PubMed1996

Neuroimmune mechanisms in health and disease: 1. Health.

A novel scientific discipline that examines the complex interdependence of the neural, endocrine and immune systems in health and disease has emerged in recent years. In health, the neuroimmunoregulatory network is fundamental to host defence and to the transfer of immunity to offspring; the network also plays important roles in intestinal physiology and in tissue regeneration, healing and reproduction. The proliferation of lymphocytes in primary lymphoid organs (bone marrow, bursa of Fabricius [in birds] and thymus) and in secondary lymphoid organs (spleen, lymph nodes and mucosal lymphoid tissue) depends on prolactin and growth hormone. These hormones allow immune cells to respond to antigen and to soluble mediators, called cytokines. Immune-derived cytokines are capable of inducing fever and of altering neuro-transmitter activity in the brain and hormone secretion by the pituitary gland. The activation of the hypothalamus-pituitary-adrenal axis by cytokines leads to immunosuppression. Lymphoid organs are innervated, and tissue mast cells respond to neurologic stimuli. In general, acetylcholine and substance P exert immunostimulatory and proinflammatory effects, whereas epinephrine and somatostatin are immunosuppressive and anti-inflammatory. In this article, the authors predict that novel approaches to immunomodulation will be possible by altering the level or efficacy of immunoregulatory hormones and neurotransmitters.

Humans↗

PubMed1996

Graft-infiltrating cells in rats receiving orthotopic semiallogeneic small intestine transplantation with portal or systemic venous drainage.

The effect of alterations in venous drainage, from either ivc to portal vein (pv), along with peritransplant systemic (ivc) or portal (pv) venous alloimmunization with irradiated semiallogenic cells, on cell subset recovery in lymphoid organs of Lewis rats receiving orthoptic small bowel allografts (from LewisXBrown Norway) F1, LBNF1) was examined. Combined portal, venous drainage and alloimmunization has been reported to increase graft/recipient survival in this model. FACS analysis using monoclonal antibodies specific for different lymphocyte subsets was performed on cell suspensions of peripheral (P) and mesenteric (M) lymph node (LN), small bowel intraepithelial lymphocytes (SBIEL), and Peyer's patch (PP) lymphocytes on days 2 and 8 posttransplantation. Donor cell contributions to these cellular analyses were estimated by comparison of FACS staining with polyclonal anti-Lewis or Lewis anti-LBNF1 antibodies. Control animals received syngeneic grafts. In both syngeneic and semi-allogenic transplants with pv or ivc drainage there was no consistent difference in cell subsets from in PLN compared with those of control nongrafted rats. Approximately 50% to 60% of these cells were alphabetaTcR+ with a CD4+/CD8+ ratio of 3-4:1 and a (CD4++CD8+)/alphabetaTcR+ ratio of 1:1. Some 5% to 12% ED3+ cells were also present. In IEL, MLN, and PP by contrast, there were significant differences in cells recovered from rats with ivc vs. pv drainage of grafts. The most striking changes reflected a decreased CD4+/CD8+ and alphabetaTcR+gammadeltaTcR+ cells in these tissues in rats predestined to show prolongation of allograft survival (ivc vs. pv injected IEL CD4/CD8+ ratios and alphabetaTcR+gammadeltaTcR+ ratios 1.0, 0.7 and 5.0, 1.0, respectively. These data are consistent with a proposed role for such gammadeltaTcR+ cells in the local regulation of graft rejection.

Animals↗

PubMed1996

A role for gamma(delta)TCR+ cells in regulation of rejection of small intestinal allografts in rats.

Lewis (LEW) rats received (Lewis x Brown Norway)F1 (LBNF1) small intestinal allografts (SIT) with graft venous drainage to either the portal vein (pv) or inferior vena cava (iv), along with immunization (pv or iv) with irradiated LBNF1 spleen cells. As reported earlier, in rats with pv drained grafts there was an increase in gammadeltaTCR+ cells infiltrating the Peyer's patches (PP) and mesenteric lymph node (MLN) compared with iv drained grafts. After restimulation in culture with irradiated LBNF1 spleen stimulator cells these PP and MLN cells from SIT rats with pv graft drainage were a prominent source of TGFbeta, IL-4, and IL-10. When subpopulations of cells from PP preparations were analyzed, an enriched (<2%betaTCR+) gammadeltaTCR+ population from SIT rats with pv graft drainage, but not iv drainage, was detected that suppressed in vitro type-1 cytokine production (IL-2, IFNgamma) from alphabetaTCR+ (<2%gammadeltaTCR+) cells derived from the MLN or peripheral lymph nodes (PLN) of these same animals. On adoptive transfer to naive LEW rats simultaneously receiving LBNF1 SIT, gammadeltaTCR+ enriched PP cells from these primary donors (pv immunized, SIT rats with pv graft drainage) produced prolonged graft/ animal survival compared with PP cells obtained from primary donors that had iv drained grafts. In addition, simultaneous infusion of anti-gammadeltaTCR monoclonal antibody into SIT rats with pv graft drainage blocked the graft enhancement normally seen in these animals. These data are consistent with an important role for type-2 cytokine producing gammadeltaTCR+ cells in the regulation of graft rejection in this model.

Animals↗

PubMed1996

Cloning and characterization of murine endothelial constitutive nitric oxide synthase.

Complementary DNA clones encoding mouse endothelial constitutive nitric oxide synthase (ecNOS) were isolated by plaque hybridization from a murine fetal cardiac lambda ZAP II cDNA expression library using a full-length human ecNOS cDNA as the hybridization probe. DNA sequence analysis indicates a 1202 amino acid protein showing significant sequence identity with human as well as bovine ecNOS.

Amino Acid Sequence↗

PubMed1996

Gamma delta TCR+ hybridomas derived from mice preimmunized via the portal vein adoptively transfer increased skin allograft survival in vivo.

C57BL/6 mice receiving pretransplant immunization with C3H.SW spleen cells via the portal vein, but not the vena cava, show Ag-specific delayed rejection of allogeneic C3H.SW skin grafts. This delayed rejection is not seen if preimmunization is performed in gamma delta TCR knockout (C57BL/6-Tcrdtm1Mom) mice. gamma delta TCR+ and alpha beta TCR+ hybridoma cells were prepared from Peyer's patch cells harvested from C57BL/6 mice 4 days following portal venous immunization with 100 x 10(6) irradiated C3H.SW spleen cells and skin grafting with C3H.SW tail skin. After recloning, these hybridoma cells were tested for cytokine production in vitro following restimulation with irradiated C3H.SW spleen cells and for their ability to delay rejection of C3H.SW skin grafts after adoptive transfer to C57BL/6 mice. Delayed graft rejection was a function of cells that showed preferential production of IL-10, not IFN-gamma, in vitro, independent of the source (vena cava or portal vein immunized mice) or the TCR phenotype of the hybridoma. Simultaneous infusion of anti-IL-10 mAb abolished this graft prolongation effect of transferred gamma delta TCR+ hybridomas. Hybridoma cells producing IL-10 on restimulation could polarize cytokine production from freshly stimulated mesenteric lymph node away from production of IL-2 and IFN-gamma, and toward IL-4, IL-10, and TGF-beta production. This immunoregulation by hybridoma cells in vivo and in vitro was observed even for third party Ag-stimulated mice/cells as long as the hybridoma cells themselves received stimulation with their specific Ag.

Adoptive Transfer↗

PubMed1996

Resistance of naive mice to murine hepatitis virus strain 3 requires development of a Th1, but not a Th2, response, whereas pre-existing antibody partially protects against primary infection.

Murine hepatitis virus strain 3 (MHV-3) produces a strain-dependent spectrum of disease. The development of liver necrosis has been shown to be related to production of a unique macrophage procoagulant activity (PCA), encoded by the gene fgl-2, in susceptible mice. These studies were designed to examine the influence of Th1/Th2 cells on resistance/susceptibility and production of macrophage PCA in resistant (A/J) and susceptible (BALB/cJ) strains of mice following infection with MHV-3. Immunization of A/J mice with MHV-3 induced a Th1 cellular immune response, and one Th1 cell line (3E9.1) protected susceptible mice and inhibited PCA production by macrophages both in vitro and in vivo. In contrast, immunization of BALB/cJ mice with an attenuated variant of MHV-3 derived from passaging MHV-3 in YAC-1 cells resulted in a Th2 response. Transfer of spleen cells and T cell lines from immunized BALB/cJ mice failed to protect naive susceptible syngeneic mice from infection with MHV-3 and augmented macrophage PCA production to MHV-3 in vitro. However, serum from immunized BALB/cJ mice contained high titrated neutralizing Ab that protected naive BALB/cJ animals from lethal primary MHV-3 infection. These results demonstrate that susceptible BALB/cJ mice generate a Th2 response following MHV-3 infection and that these Th2 cells neither inhibit MHV-3-induced macrophage PCA production nor protect naive mice from MHV-3 infection. The results suggest that Ab protects against primary infection but cannot eradicate ongoing infection. Thus, these data define the differential role of Th1/Th2 lymphocytes in primary and secondary MHV-3 infection and emphasize the importance of PCA in the pathogenesis of MHV-3 infection.

Animals↗