Chronic Graft-versus-host Reaction Research Articles

CD28 knockout mice as a useful clue to examine the pathogenesis of chronic graft-versus-host reaction

CD28 knockout mice as a useful clue to examine the pathogenesis of chronic graft-versus-host reaction.BackgroundInjection of BALB/c or DBA/2 spleen cells into F1 C57BL/6 (B6) hybrids induces a graft-versus-host reaction (GVHR) of a chronic stimulatory type that results in clinical and pathologic manifestations that resemble the human systemic lupus erythematosus (SLE). The aim of the present study was to examine the role of a major T-cell costimulatory signal receptor, CD28, in the production of autoantibody and the development of an immune complex glomerulonephritis, which are common in SLE pathology.MethodsFor this purpose, CD28-deficient (CD28KO) mice were used for the source of donor lymphocytes. Chronic GVHR was induced by an injection of BALB/c or BALB. CD28KO donor cells into normal BCF1 mice. Serum titers of anti-dsDNA antibodies were assessed by enzyme-linked immunosorbent assay (ELISA) and major histocompatibility complex (MHC) class II antigen expression on B cells were tested by flow cytometry. In addition, depositions of immunoglobulin (Ig) were examined by direct immunofluorescence staining on frozen kidney sections.ResultsWhen (BALB/c × B6)F1 mice were injected with parental BALB/c lymphocytes, serum anti-dsDNA titer was significantly increased in association with nonspecific B-cell activation and IgG deposition in the glomerular basement membrane. In sharp contrast, none of these signs were observed in F1 mice, which were injected with CD28KO spleen cells.ConclusionThe CD28-mediated T-cell costimulatory pathway plays a pivotal role in the development of polyclonal B-cell activation, autoantibody production, and an immune complex glomerulonephritis. We propose that CD28KO mice are useful clues in examining the pathogenesis of experimental lupus nephritis. CD28 knockout mice as a useful clue to examine the pathogenesis of chronic graft-versus-host reaction. Injection of BALB/c or DBA/2 spleen cells into F1 C57BL/6 (B6) hybrids induces a graft-versus-host reaction (GVHR) of a chronic stimulatory type that results in clinical and pathologic manifestations that resemble the human systemic lupus erythematosus (SLE). The aim of the present study was to examine the role of a major T-cell costimulatory signal receptor, CD28, in the production of autoantibody and the development of an immune complex glomerulonephritis, which are common in SLE pathology. For this purpose, CD28-deficient (CD28KO) mice were used for the source of donor lymphocytes. Chronic GVHR was induced by an injection of BALB/c or BALB. CD28KO donor cells into normal BCF1 mice. Serum titers of anti-dsDNA antibodies were assessed by enzyme-linked immunosorbent assay (ELISA) and major histocompatibility complex (MHC) class II antigen expression on B cells were tested by flow cytometry. In addition, depositions of immunoglobulin (Ig) were examined by direct immunofluorescence staining on frozen kidney sections. When (BALB/c × B6)F1 mice were injected with parental BALB/c lymphocytes, serum anti-dsDNA titer was significantly increased in association with nonspecific B-cell activation and IgG deposition in the glomerular basement membrane. In sharp contrast, none of these signs were observed in F1 mice, which were injected with CD28KO spleen cells. The CD28-mediated T-cell costimulatory pathway plays a pivotal role in the development of polyclonal B-cell activation, autoantibody production, and an immune complex glomerulonephritis. We propose that CD28KO mice are useful clues in examining the pathogenesis of experimental lupus nephritis.

CD28 knockout mice as a useful clue to examine the pathogenesis of chronic graft-versus-host reaction

Injection of BALB/c or DBA/2 spleen cells into F1 C57BL/6 (B6) hybrids induces a graft-versus-host reaction (GVHR) of a chronic stimulatory type that results in clinical and pathologic manifestations that resemble the human systemic lupus erythematosus (SLE). The aim of the present study was to examine the role of a major T-cell costimulatory signal receptor, CD28, in the production of autoantibody and the development of an immune complex glomerulonephritis, which are common in SLE pathology. For this purpose, CD28-deficient (CD28KO) mice were used for the source of donor lymphocytes. Chronic GVHR was induced by an injection of BALB/c or BALB. CD28KO donor cells into normal BCF1 mice. Serum titers of anti-dsDNA antibodies were assessed by enzyme-linked immunosorbent assay (ELISA) and major histocompatibility complex (MHC) class II antigen expression on B cells were tested by flow cytometry. In addition, depositions of immunoglobulin (Ig) were examined by direct immunofluorescence staining on frozen kidney sections. When (BALB/c x B6)F1 mice were injected with parental BALB/c lymphocytes, serum anti-dsDNA titer was significantly increased in association with nonspecific B-cell activation and IgG deposition in the glomerular basement membrane. In sharp contrast, none of these signs were observed in F1 mice, which were injected with CD28KO spleen cells. The CD28-mediated T-cell costimulatory pathway plays a pivotal role in the development of polyclonal B-cell activation, autoantibody production, and an immune complex glomerulonephritis. We propose that CD28KO mice are useful clues in examining the pathogenesis of experimental lupus nephritis.

Modulation of the immunoglobulin dysregulation in GvH- and SLE-like diseases by the murine IL-4 receptor (IL-4-R)

As has been reported previously, models of chronic graft-versus-host (GvH) and systemic lupus erythematosus (SLE)-like diseases are characterized by high IgE and IgG1 immunoglobulin (Ig) levels in the serum. An IL-4 induced pathological expansion of Th2 helper cells has been described for both disease models. Due to the immunopharmacological profile of soluble recombinant interleukin-4 receptor (IL-4-R) to bind specifically the corresponding ligand IL-4 and thereby to modulate biological activity upon exogenous administration in various autoimmune disease models, we investigated the immunoregulatory activity of IL-4-R and anti-IL-4 monoclonal antibody (MAb) 11B11 on the development of SLE-like disease in MRL/lpr autoimmune mice and on chronic GvH reaction in BDF1 hybrid mice. Sensitized GvH-BDF1 hybrid mice and SLE in MRL/lpr autoimmune mice were treated in vivo with the IL-4 antagonists to alter the pattern of serum Ig production and to modulate the disease process. These animals were followed for proteinuria, autoantibody production (anti-dsDNA), serum IgE, IgG1 and IgG2a levels, and the survival was monitored. Treatment of these diseased animals resulted in an improved survival rate, lowered the percentage of animals with lymphadenopathy and hepatosplenomegaly, reduced the levels of autoantibodies and inhibited proteinuria of the developing glomerulonephritis in both mouse strains, even in the established diseases. In both models the increase in total IgE and IgG1 levels in serum was strongly inhibited by the IL-4 antagonists, even under therapeutic conditions. But there was no inhibitory activity observed on the IgG2a serum levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Suppression of Local and Systemic GVHR by Supernatants from Human Choriocarcinoma Cell Lines

Graft versus host reaction (GVHR) and host versus graft reactivity during pregnancy share at least some cellular pathways. Immunosuppression mediated by placental suppressor factors may actively contribute to the survival of the conceptus in the immunocompetent mother. We have previously reported that supernatants from human choriocarcinoma cell lines inhibit mitogen-activated and antigen-activated proliferation of lymphoid cells. From this perspective, we investigated the capacity of supernatants from human choriocarcinoma cell lines (HCS) to suppress in vivo graft versus host reactions in mice. HCS was tested for its ability to block both local as well as systemic GVHR induced by the injection of parental cells into F1 mice. HCS mediates profound suppression of acute and chronic GVHR. These results confirm that human choriocarcinoma cell lines secrete a potent immunosuppressive factor(s) which can efficiently modulate immune responses in vivo.

Polyclonal B Cell Activation and Antigen-Specific Stimulation in Mice with Chronic Graft-versus-Host Reaction

Primary chronic graft-versus-host reaction (GVHR) was induced in (C57BL/10 × DBA/2)F 1 (BDF 1) mice via the transfer of parental DBA/2 derived lymphocytes. A significant increase in the absolute number of IgM- and IgG-secreting cells was found in recipient BDF1 mice at 3 weeks post-transfer. The increase included B cells reactive with both self- and non-self-antigens, without being skewed toward the production of autoantibodies. Later in the disease process (approximately 6-8 weeks post-transfer), we detected an increased number of B cells that secreted antibodies reactive with a select subset of autoantigens in about 30% of BDF 1 recipients (3 of 9 mice). These findings suggest that not only antigen-specific stimulation, which was detected in a late stage, but also initial polyclonal activation may account for chronic GVHR.

Thymus: a direct target tissue in graft-versus-host reaction after allogeneic bone marrow transplantation that results in abrogation of induction of self-tolerance.

Graft-versus-host reaction (GVHR) following allogeneic bone marrow (BM) transplantation was investigated by analyzing expression of antigen receptors on T cells specific for recipient antigens. GVHR chimeras were prepared by transplanting mixtures of splenic T cells and T-cell-depleted BM cells from B10 (I-E-, Mls-1b) or B10.AQR (I-E+, Mls-1b) mice into lethally irradiated AKR/J (I-E+, Mls-1a) recipients. Increased proportions of V beta 6+ T cells reactive to recipient antigens (I-E and Mls-1a) were observed in thymuses from such chimeras 1 or 5 wk after BM transplantation. V beta 6+ T cells observed 1 wk after BM transplantation were derived from mature T cells that had been inoculated into recipients. These cells responded to recipient antigens expressed in the thymus. After 5 wk, thymocytes brightly positive for V beta 6+ were shown not to descend from mature T cells but to differentiate from precursor cells present in the BM inocula. Since V beta 6+ T cells were eliminated in thymuses from non-GVHR chimeras 5 wk after BM transplantation using T-cell-depleted BM cells alone, it appears that GVHR occurring in the thymus at an early stage abrogates thymic stromal functions essential to induction of self-tolerance in the T-cell repertoire. These findings propose a mechanism (autoimmunity) to explain in part the pathogenesis of chronic GVHR.

Cutaneous Graft-Versus-Host Reaction Lacks Evidence of Cutaneous Cytomegalovirus by the Immunoperoxidase Technique

Sixty skin biopsy specimens from 21 bone-marrow transplant patients were evaluated for the presence of cytomegalovirus (CMV) using two monoclonal antibodies to early and late antigens. Each patient had at least one biopsy showing an acute graft-versus-host reaction (GVHR), grade 2, and one positive culture for CMV from blood, bone marrow or urine. In no case could CMV antigens be identified in biopsies showing an acute or chronic cutaneous GVHR or in any other of the skin biopsies obtained from these patients. While CMV may play a role in immunologic events culminating in graft-versus-host disease (GVHD), this immunoperoxidase study did not reveal evidence of viral antigens in tissue displaying features of cutaneous GVHR.

Graft-Versus-Host Reactions in F 1 Hybrid Mice: MHC-Restriction-Independent Generalized Depression of Virus-Specific Cytotoxic T Cell Response

A graft-versus-host reaction (GVHR) was induced in F 1 hybrid mice by intravenous injection of parental spleen cells. A possible effect generated by recipient F 1 mice of a GVHR on the restriction specificities of anti-viral cytotoxic T cells was investigated. (A/J × C57BL/6) F 1 recipients were injected with 1 × 10 8 or 2 × 10 7 spleen cells from either parent. Zero to two weeks later during an acute GVHR or 12 to 26 weeks later during a chronic GVHR, spleen cells from these F1 recipients were assayed for their capacity to generate vaccinia (or lymphocytic choriomeningitis) virus-specific cytotoxic T cells. No effect was seen when parental cells were injected with virus on the same day. During the acute phase of the GVHR, recipients of spleen cells of both doses or from either parent injected 7 or 14 days previously generated markedly fewer cytotoxic T cells with respect to both parental restriction specificities. In the more chronic situation only, recipients of 1 × 10 8 parental spleen cells showed depressed virus-specific cytotoxic responses, again both restriction specificities were affected comparably. Therefore, a GVHR depresses generation of virus-specific cytotoxic T cells in a dose- andtime-dependent way, but does not measurably disturb the balance of parental 1 versus parental 2 restriction specificity in an F 1 recipient.

The graft-versus-host reaction and immune function. I. T helper cell immunodeficiency associated with graft-versus-host-induced thymic epithelial cell damage.

The injection of parental A strain lymphoid cells into adrenalectomized CBAxA F1 (BAF1) mice induced a chronic graft-versus-host (GVH) reaction resulting in T cell and B cell immunosuppression as well as thymic epithelial cell injury, but not stress-related thymic involution. Thymocytes from BAF1 mice undergoing a GVH reaction were studied for their ability to reconstitute T helper cell (TH) function and phytohemagglutinin (PHA) and concanavalin A (Con A) mitogen responses in thymectomized, irradiated, BAF1 mice reconstituted with normal syngeneic bone marrow (ATxBM). Thymocytes from BAF1 mice early after the induction of a GVH reaction (days 10-12) were as effective as normal thymocytes in reconstituting TH and mitogen responses. Thymocytes from BAF1 mice 40 or more days after the induction of a GVH reaction did not reconstitute either the TH function or PHA and Con A responses in ATxBM mice. The inability to reconstitute ATxBM mice was not due to the presence of suppressor cells contained in the thymocyte inoculum. It is proposed that GVH-induced thymic epithelial cell injury blocks or arrests normal T cell differentiation, resulting in a population of thymocytes that lack the potential to become competent T helper cells or mitogen-responsive cells when transferred into ATxBM mice. This thymic functional defect results in a permanent TH immunodeficiency in mice experiencing a chronic GVH reaction.

Lichen planus and chronic graft-versus-host reaction. In situ identification of immunocompetent cell phenotypes.

The purpose of the present study was to examine the phenotype of the cutaneous immunocompetent cells in lichen planus and chronic graft versus host (GVH) reaction infiltrates, by the use of monoclonal antibodies directed against T cell populations and Langerhans cells. Our results in lichen planus suggest an immunological reaction similar to the delayed hypersensitivity reaction, including all the immunocompetent cell subpopulations, with a first stage of antigenic information by Langerhans cells (OKT6+, BL6+, HLA-DR+) and helper cells, and a second stage mediated by suppressor/cytotoxic cells. The results from the study of GVH reaction also suggest an effect mediated by suppressor/cytotoxic cells (OKT3+, OKT4-, OKT8+, HLA-DR+). Our results favour the existence of a lymphocytotoxic process in lichen planus and chronic GVH reaction.

703 PROGRESSIVE SYSTEMIC SCLEROSIS (PSS)-LIKE SYNDROME FOLLOWING BONE MARROW TRANSPLANTATION

A 15 year old boy developed skin changes compatible with chronic graft versus host reaction (CGVHR) following bone marrow transplantation. His skin was thin, dry, inelastic, and telan-giectatic. Biopsy revealed basal vacuolization, loss of rete pegs, atrophy of sweat glands, perivascular lymphocyte cuffing, and dense dermal collagenization. Because of the dermatologic similarity to PSS, we compared this patient to 38 PSS patients for evidence of systemic involvement. He had significant restrictive lung disease (VC and TLC 50% of predicted), cardiac conduction abnormalities (first degree heart block, right bundle branch block), and a pericardial effusion. Each of these abnormalities was found in 30-40% of the PSS group. Arteriolar intimal proliferation was seen in the heart and kidneys of our patient at autopsy. This finding has been noted in approximately 25% of PSS patients. Our patient had Raynaud's phenomenon as did 95% of the PSS group. He had a striking polyclonal elevation in serum IgG (4 gms/dl), an uncommon finding in PSS. 84% of the PSS patients showed esophageal dysmotility, but our patient did not have this abnormality. The clinical and pathologic similarities between this patient and PSS patients suggest that CGVHR and PSS may have a common pathogenesis. The use of animal models of graft versus host reaction may lead to an understanding of mechanisms responsible for both disease processes.

The induction of murine neoplasia

<h2>Summary</h2> The hypotheses suggesting that a chronic form of the graft-versus-host reaction (GvHR) or minor histocompatibility differences in lymphoid cells are sufficient to produce murine neoplasia, have been tested in mice with a low natural incidence of murine lymphoma/leukaemia. There was no evidence of an increased incidence of neoplasia in the mice tested. The development of the natural levels of leukaemia was not brought forward to an earlier age as a result of the initiation of a chronic GvHR. Neither was the incidence of leukaemia increased nor brought forward in a system where minor histocompatibility differences existed between host and donor lymphoid cells. These results are considered in relation to the views that chronic immunological injury, with or without virus, may give rise to neoplasia in the mouse.