Biomedical subjects

W S Lapp

Publications and source records attributed to W S Lapp.

At least 37 records · Page 2Linked to original sources

PubMed1991

Induction, specificity and elimination of asialo-GM1+ graft-versus-host effector cells of donor origin.

In previous studies we demonstrated that an induced asialo-GM1 positive (ASGM1+) cell of donor origin that exerts natural killer cell-like activity (NK activity+) plays a crucial role in the development of graft-versus-host (GVH)-associated tissue damage and severe immunosuppression. This study examined whether the ASGM1+ (NK activity+) GVH effector cells were activated by non-specific signals or whether these cells were triggered by specific alloantigens and displayed antigenic specificity. C57B1/6 (B6) donor mice were treated with either B6 x AF1 (B6AF1) lymphoid cells and anti-asialo GM1 antibodies (anti-ASGM1) to induce and eliminate specifically activated B6-anti-B6AF1 ASGM1+ (NK activity+) cells or with polyinosinic: polycytidylic acid (poly I:C), and anti-ASGM1 to eliminate non-specifically activated ASGM1+ (NK activity+) cells. Donor spleen and lymph node cells depleted of the specific allo-induced ASGM1+ NK reactive cells showed near normal numbers of L3T4+ and Lyt-2+ cells and retained T- and B-cell functions as measured by mitogen responses (to PHA, Con A and LPS), mixed lymphocyte responses (MLR) (to B6AF1) and the generation of cytotoxic T cells (CTL) (to B6AF1 blasts). Anti-ASGM1 treatment almost completely abrogated NK activity in all donor inocula. GVH reactions were induced by injecting treated donor cells into B6AF1, B6 x C3HejF1 (B6C3HF1) and B6 x SJLF1 (B6SJLF1) hybrids and monitored by splenomegaly, suppression of T-cell mitogen responses and the development of histopathological lesions in the thymus, liver and pancreas. Cells from donors depleted of non-specifically (poly I:C) induced ASGM1+ cells induced severe histological lesions, marked immunosuppression and splenomegaly in all three F1 hybrid combinations. When the donor cells were depleted of specifically induced (B6-anti-B6AF1) ASGM1+ cells and injected into the three F1 combinations they induced splenomegaly in all three but caused severe tissue injury and intense immunosuppression only in B6C3HF1 and B6SJLF1 mice and not in B6AF1 mice. Genetic analysis suggests that the H-2D (or a closely related) region of the H-2 complex plays an important role in the activation of the specific GVH effector cells. These results suggest that the cell(s) responsible for splenomegaly are different from the ones that cause severe GVH-associated tissue damage and immunosuppression although there may be cells and/or lymphokines common to both processes.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1991

Expression of tumor necrosis factor alpha in the developing nervous system.

We present evidence that tumor necrosis factor alpha (TNF-alpha) is transiently expressed at specific times during embryogenesis in precisely defined areas of the nervous system in two different classes of vertebrates. In murine embryos, TNF-alpha was detected in the brain, neural tube and peripheral mixed spinal nerves. In the chick embryo, TNF-alpha was observed in the brain neuroepithelium and in the developing Purkinje neurons of the cerebellum. Western immunoblot analysis revealed that brain tissue from both mouse and chick embryos contained a 50 kDa protein showing immunoreactivity with anti-TNF-alpha antibody. These results suggest that TNF-alpha participates in the normal development of the vertebrate brain and spinal cord.

Animals↗

PubMed1990

Lipopolysaccharide-induced fetal resorption in mice is associated with the intrauterine production of tumour necrosis factor-alpha.

Certain strains of mice display an increased frequency of fetal resorption, but little is known about the effector mechanisms involved. We have examined the events associated with lipopolysaccharide (LPS)-induced fetal resorption in mice. Administration of 25 micrograms LPS on Day 12 of gestation resulted in the appearance of tumour necrosis factor-alpha (TNF-alpha) in the amniotic fluid and fetal resorption. Levels of LPS-induced TNF-alpha were reduced by 90% after pretreatment with the TNF-alpha-suppressing drug pentoxifylline (PXF). Treatment of pregnant mice during early gestation with 0.1 micrograms LPS resulted in fetoplacental resorption which was maximal when the LPS was given on Day 8. Resorption induced by 0.1 micrograms LPS on Day 8 of gestation was significantly reduced by pretreatment with PXF. Infiltration of asialo-GM1-positive cells was observed in the decidual-ectoplacental cone area of embryonic units from LPS-treated mice. In addition, treatment with anti-AGM1 antiserum prevented the LPS-induced resorption. Our results suggest that TNF-alpha and asialo-GM1-positive cells are involved in LPS-induced fetal resorption.

Amniotic Fluid↗

PubMed1989

The effects of polyinosinic:polycytidylic acid on the graft-versus-host reaction. III: Increased severity of the reaction with delayed pI:C treatment.

We have been investigating the effects of polyinosinic:polycytidylic acid (pI:C), an interferon inducer, on the graft-versus-host reaction. We have previously shown that pI:C treatment of C57BL/6xAF1 (B6AF1) recipient mice immediately before injection of C57BL/6 (B6) parental lymphocytes inhibited the immuno-suppression and pathological changes normally caused by the GVH reaction, by a mechanism apparently identical to that seen in F1 hybrid resistance (HR) to hematopoietic grafts. We now demonstrate that delaying pI:C treatment by as little as 48 hr produces the opposite effect. Treatment of recipient B6AF1 mice at different days after transfer of parental lymphocytes induced a marked increase in the severity of the GVH reaction, as measured by a decreased plaque-forming cell response to sheep erythrocytes; decreased proliferative response to the T and B cell mitogens PHA, Con A, and LPS; increased pathological changes in both lymphoid and nonlymphoid tissues; and increased GVH-associated mortality. This effect is unrelated to HR, as pI:C was able to augment the severity of the GVH reaction when A strain cells were injected into AxCBAF1 recipients, which do not manifest HR. Early pI:C treatment (1 and 2 days after parental cell transfer) increased the severity of the GVH reaction much more than later pI:C treatment (7 and 8 days after parental cell transfer). This observation, along with the demonstration of altered pathology in GVH mice treated with pI:C, suggests that the effect of pI:C is not mediated through a direct suppressive effect of IF on the cells responding in either the PFC or mitogen assays, but rather by the ability of IF to activate or suppress mechanisms involved in the development of GVH-induced alterations.

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PubMed1988

Association between the degree of thymic dysplasia and the kinetics of thymic NK cell activity during the graft-versus-host reaction.

In this study, by employing different cell doses and parent into F1 hybrid combinations, we have investigated the relationship between the severity of thymic medullary dysplasia and the kinetics of thymic natural killer (NK) cell activity after the induction of graft-versus-host (GVH) reactions. GVH reactions were induced by injecting different doses (30, 20, 10 X 10(6] of C57BL/6 (B6) of A parental lymphoid cells (PLC) into non-X-irradiated adult B6xAF1 (B6AF1) mice. On different days after the induction of GVH reactions, the thymuses were examined histologically and thymocyte NK cell activity was tested by using YAC targets. Our results show that, depending upon the genotype and dose of PLC injected, various degrees of thymic medullary dysplasia (mild, moderate, or severe) can be induced. Furthermore, severe to moderate thymic medullary dysplasia is observed only in those groups of GVH-reactive mice in which thymic NK cell activity occurs early and increases rapidly. In contrast, when mild thymic medullary dysplasia or no thymic alterations was observed, thymic NK cell activity peaked later and was of lower intensity than that of the groups with moderate to severe lesions. These results suggest an association between the degree of thymic medullary dysplasia and the kinetics of NK cell activity in the thymus. Furthermore, the different degrees of thymic medullary dysplasia as described here may serve as a powerful tool to study the role of thymic medullary dysplasia in determining the duration of T-cell immunodeficiency associated with the GVH reactions.

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PubMed1988

Prevention of murine graft-versus-host disease by inducing and eliminating ASGM1+ cells of donor origin.

In this study anti-asialo GM1 antibodies (anti-ASGM1) were used to further characterize the effector cells responsible for graft-versus-host (GVH)-induced histopathological lesions: Two different types of ASGM1+ cells were identified: an endogenous ASGM1+ population and an induced ASGM1+ population. Both of the ASGM1+ cell populations exhibited natural killer (NK) cell activity, as assessed by their ability to lyse YAC tumor targets in vitro. Donor C57BL/6 (B6) mice were treated in vivo with anti-ASGM1 to eliminate endogenous ASGM1+ cells. ASGM1+ cells were induced in B6 donor mice by treating the animals with 15 x 10(6) B6 x AF1 (B6AF1) lymphoid cells for 44-48 hr. The induced ASGM1+ cells were eliminated by in vivo treatment with anti-ASGM1. GVH reactions were induced by injecting B6 lymphoid cells into B6AF1 mice. Prior to GVH induction the B6 donor cells were tested for NK cell activity against YAC tumor target cells in vitro and for T and B cell functions by mitogen responses in vitro. GVH reactions were determined by splenomegaly, suppression of the plaque-forming cell (PFC) response to sheep red blood cells (SRBC), suppression of the T and B cell mitogen responses, and the development of GVH-associated histopathological alterations in the thymus, liver, and pancreas. Donor lymphoid cells depleted of endogenous ASGM1+ cells were effective at inducing splenomegaly, severe suppression of immune functions, and histopathological lesions. Donor lymphoid cells depleted of both the endogenous and induced ASGM1+ cells displayed normal T cell mitogen responses and were capable of inducing splenomegaly and partial suppression of the PFC response to SRBC when injected into B6AF1 recipients, however, these lymphoid cells failed to induce both GVH-associated histopathological lesions and severe suppression of T and B cell mitogen responses. These results suggest that semiallogeneic stimulation induces an ASGM1+ population in the donor inoculum that displays NK cell-like function (YAC killing) and that plays a crucial role in inducing GVH-mediated histopathological lesions and severe immunosuppression of both T and B cell responses.

Animals↗

PubMed1988

Genetics of resistance to infection with Candida albicans in mice.

To determine differences in susceptibility, 234 naive mice including xid and beige mutants were infected intravenously with Candida albicans and monitored with survival analysis and quantitative culture of the kidneys. By using survival time as the criterion, animals of seven inbred strains were separated into three groups. C3H/HeJ and Dw/+ were most susceptible; C57BR/cdJ, BRVR and CBA/N (xid) were intermediate in susceptibility; C57BL/KsJ and C57BL/6J were least susceptible. Mean survival times (MST) were markedly influenced by the number of Candida cells injected while the ranking of mouse strains by survival alone was unchanged. There was a dissimilar behaviour of the strains to produce organ weight changes in response to infection when compared with uninfected mice which were matched for age and genetic lineage. Black mice had lower colony forming units (CFU) per mg of tissue at the time of death than animals of other genetic lineage. Nevertheless, the finding that MST and CFU studies were loosely correlated in a few strains of mice indicated that the proliferation of the fungus in the kidneys was not always the major cause of death. The beige mutation was found to determine an increased susceptibility to systemic Candida infection. The differences in survival for beige and nonbeige mice were influenced by the genetic lineage of the host, being much greater in the C57BL/6 strain (36.7 days) than in the C3H/He strain (5 days). C57BL/6 beige-J had significantly higher CFU per organ and per unit of weight than C57BL/6 +/+ mice. These data evinced an important contribution of host genetic factors to resistance to systemic candidiasis. It is suggested that innate resistance genes regulate the differentiation in the bone marrow and the function of cells of granulocyte-macrophage lineage.

Animals↗

PubMed1988

Complete sequential regeneration of graft-vs.-host-induced severely dysplastic thymuses. Implications for the pathogenesis of chronic graft-vs.-host disease.

This study presents the sequential morphologic regeneration of graft-vs.-host (GVH)-induced dysplastic thymuses in long-term survivors of GVH reactions. GVH reactions were induced in adult C57BL/6xAF1 (B6AF1) hybrids by injecting 20 x 10(6) A strain parental lymphoid cells (PLC). Starting on day 30 after GVH induction, five to ten animals were randomly selected from a pool of GVH-reactive mice and killed at various times. Each animal was tested for thymic histology and T cell functions. Thymuses taken on day 30 after GVH induction displayed severe dysplasia as characterized by lymphocytic depletion, complete effacement of cortico-medullary demarcation, and reduction and total loss of medullary epithelial cells or both. Starting by days 60-70 after GVH induction, at least four stages of thymic regeneration were identified. Day 60-70 thymuses displayed cortical regeneration and the reappearance of cortico-medullary demarcation. The medulla of these thymuses, although containing dark individual epithelial cells and numerous lymphocytes, was devoid of pale epithelial cells (stage 1). The medulla of thymuses on day 100 after GVH induction displayed a few sparcely distributed pale epithelial cells and numerous lymphocytes as well as dark epithelial cells (stage 2). The medulla of thymuses examined 130 days after GVH induction displayed numerous pale individual epithelial cells and a few pale epithelial cell clusters. Such thymuses also showed a reduction in the number of medullary lymphocytes (stage 3). Finally, the medulla of thymuses 150-160 days after GVH induction displayed numerous pale epithelial cell clusters and Hassall's bodies. These thymuses were indistinguishable from normal adult thymuses (stage 4). All of the animals tested up to day 130 after GVH induction showed no significant T cell function. Animals displaying stage 4 of thymic regeneration showed significant proliferative responses to T cell mitogen, concanavalin A (conA), and six of ten animals also displayed a few plaque forming cells (PFC) to sheep red blood cells (SRBC) in their spleens. Furthermore, all animals (10 of 10) killed on day 180 after GVH induction displayed significant T cell functions.(ABSTRACT TRUNCATED AT 400 WORDS)

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PubMed1987

Suppression of B lymphocyte genesis in the bone marrow by systemic graft-versus-host reactions.

The effects of systemic graft-versus-host (GVH) reactions on B lymphocyte production in the bone marrow of mice were examined by quantitating populations of pre-B cells and B lymphocytes. Acute and chronic GVH reactions were induced by injecting A strain lymphoid cells into either (C57BL/6 X A) F1 or (CBA X A) F1 mice, respectively. Control groups of F1 hybrid mice were given syngeneic lymphoid cells. By double immunofluorescence labeling for cytoplasmic mu heavy chains of IgM (c mu) and for surface mu (s mu) the absolute numbers of pre-B cells (c mu + s mu-) and B lymphocytes (s mu +) in the bone marrow and spleen were determined. During acute GVH reactions, the pre-B cells and B lymphocytes in the bone marrow fell rapidly in numbers and were almost absent from 16 days until the end of the 30-day experimental period. In the spleen, the number of B lymphocytes remained normal for 8 days, then fell to less than 2% of control values from 16 days onward. A similar initial decline in pre-B cells and B lymphocytes occurred during chronic GVH reactions. In long-term survivors of GVH reactions, pre-B cells and B lymphocytes began to reappear after 40 days and maintained normal numbers from 100 to 150 days. The antibody response of spleen cells to sheep red blood cells was lost during GVH reactions. However, this occurred even before B lymphocytes were eliminated and the response remained subnormal after B lymphocyte numbers had recovered. The results demonstrate that systemic GVH reactions markedly depress the normally active genesis of primary B lymphocytes in the bone marrow of the host, accounting in part for the associated state of humoral immunodeficiency.

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PubMed1987

Graft-versus-host reactions in the beige mouse. An investigation of the role of host and donor natural killer cells in the pathogenesis of graft-versus-host disease.

To investigate the role of natural killer (NK) cells in the induction and pathogenesis of graft-versus-host (GVH) disease, +/beige (+/bg; normal NK cell activity) and beige/beige (bg/bg; deficient NK cell activity) parental C57BL/6 (B6) lymphoid cells were used to induce GVH reactions in either B6 X C3H/Hej +/bg (+/bgF1) or B6 X C3H/HeJF1 bg/bg (bg/bg F1) hybrid mice. When B6 bg/bg parental lymphoid cells (PLC) were injected into bg/bg F1 mice, early splenomegaly, early severe suppression of the plaque-forming cell (PFC) response to sheep red blood cell (SRBC), and only partial suppression of T cell mitogen responses to concanavalin A (Con A) and phytohemagglutin (PHA) were observed on day 12 after GVH induction. In the same GVH combination, slightly augmented NK cytotoxic activity was induced and no GVH-induced moderate-to-severe pathological alterations in the liver and pancreas were observed. When bg/bg PLC were injected into +/bg F1 mice, early splenomegaly and pronounced immunosuppression of the PFC response to SRBC and partial suppression of Con A and PHA responses were observed on day 12 after GVH induction. In this combination (bg/bg----+/bg F1), significant NK cell activity was induced, but no moderate-to-severe histopathological alterations were observed. In contrast, when B6 +/bg PLC were injected into either +/bg F1 or bg/bg F1 hybrids, early splenomegaly, and severe immunosuppression of both the PFC response to SRBC and the T cell mitogen responses to Con A and PHA were observed by day 12--which persisted until day 30 after GVH induction. Furthermore, high NK cell activity was recorded and moderate-to-severe histopathological alterations appeared in both +/bg F1 and bg/bg F1 recipients. These results show that the bg/bg PLC can induce GVH-associated early splenomegaly and immunosuppression of the PFC response to SRBC in both the bg/bg F1 and +/bg F1 hybrids, but that it failed to induce moderate-to-severe histopathological alterations, even though NK cell activity of host origin was activated during GVH reactions. Conversely, when +/bg donor cells were used to induce the GVH reaction, splenomegaly and immunosuppression, as well as moderate-to-severe histopathological lesions were induced. These results suggest that donor NK cells rather than host NK cells play an active role in GVH-associated tissue damage, which in turn contributes to the long-term suppression.

Animals↗

PubMed1987

Kinetics of natural killer cell cytotoxicity during the graft-versus-host reaction. Relationship between natural killer cell activity, T and B cell activity, and development of histopathological alterations.

The relationships between splenic natural killer (NK) cell cytotoxicity, T and B cell function, and the development of histopathological lesions in the liver and pancreas have been studied during the course of graft-versus-host (GVH) reactions. GVH reactions were induced in (C57BL/6 X A)F1 (B6AF1) hybrids by different doses, (10,20 and 30 X 10(6)) of either parental strain C57/BL6 (B6) or A lymphoid cells. Splenic NK cell cytotoxicity was studied by employing YAC-1, an NK-cell-sensitive target. Splenic T and B cell function were assessed by mitogen responsiveness to concanavalin A, phytohemagglutinin, and Escherichia coli lipopolysaccharide, and by the in vitro plaque-forming cell response to sheep red blood cells. Histopathological lesions characteristic of GVH reactions were recognized at a time (day 8 after GVH induction) when both T and B cell functions were totally suppressed and NK cell activity was greatest. The severity of histopathological alterations later (day 16 after GVH induction) correlated with an early peak in NK cell cytotoxicity rather than with the overall NK cell activity. When low doses (10,20 X 10(6)) of B6 cells were employed to induce GVH reactions, a significant increase in NK cell activity was observed, yet neither histopathological alterations nor suppression of T and B cell functions were observed. The killing of YAC-1 targets by splenocytes obtained from the different GVH combinations could not be abrogated by pretreatment with anti-Thy 1.2 serum plus complement, suggesting that T lymphocytes were not central to this cytolytic process. These experiments demonstrated that: (1) an inverse relationship between T and B cell function and NK cell activity was observed early after GVH induction, (2) the severity of histopathological lesions and immunosuppression, as well as the degree of overall augmented NK cell activity, was determined by the dose and genotype of donor cells injected to induce GVH reactions, and (3) GVH-associated moderate-severe lesions occurred only in groups in which NK cell activity peaked early--whereas when NK cell activity peaked later, either mild or no lesions were observed, suggesting that the early rapid increase of NK cell activity may be useful for predicting the severity of GVH pathogenesis.

Animals↗

PubMed1986

The effects of polyinosinic:polycytidylic acid (pI:C) on the graft-vs-host (GVH) reaction. II. Increased NK-mediated rejection on C57BL/6 lymphocytes by (C57BL/6 X A)F1 mice.

We previously demonstrated that treatment of (C57BL/6 X A)F1 (F1) recipient mice with polyinosinic:polycytidylic acid (pI:C) before injection with 30 X 10(6) C57BL/6 (B6) lymphocytes prevents both the immunosuppression and pathologic lesions typical of graft-vs-host (GVH) reactions. We now report the further characterization of this phenomenon. Donor spleen and lymph node cells were labeled with fluorescein in vitro and injected into pI:C-treated or untreated mice. Two days later, recipient splenocytes were analyzed for the presence of fluorescein-labeled donor cells by flow microfluorometry. Treatment of F1 mice with pI:C resulted in a sharp reduction in the recovery of labeled B6 but not A strain parental cells. Treatment with pI:C had no effect when syngeneic recipients were used, or when F1 cells were injected into A, B6, or F1 recipients. These results suggest that pI:C treatment induces rejection of B6 but not A or F1 lymphocytes by F1 hybrid mice at least as early as 2 days after donor cell transfer. As F1 cells are not rejected by either parent, rejection does not seem to be directed against classical alloantigens. These observations are compatible with the previously described model of hybrid resistance (HR) against bone marrow grafts. The rapidity of rejection strongly suggested that natural cytotoxic mechanisms were involved, thus, natural killer (NK) cell and macrophage (M phi) cytotoxic activities were tested throughout the time when the parental cell graft was being rejected. Over this period, pI:C treatment increased cytotoxic activity against the NK-sensitive target cell line YAC-1 but had no effect on spontaneous M phi tumoricidal activity against the L5178Y and MDAY-D2 cell lines. The results suggest that NK cells, but not M phi, may be involved in the elimination of B6 parental cells by the pI:C-treated F1 mice. NK cells have been demonstrated to be radioresistant; thus, as a test of our hypothesis, we examined the effects of irradiation on the capacity of pI:C treated F1 mice to reject B6 lymphocytes. The results show that this capacity was not blocked by 750 cGy, a dose of radiation that abrogates most T and B cell functions. Furthermore, rejection of parental cells could be prevented by treatment of recipient F1 mice with antibodies to asialo GM1, a treatment that suppresses NK activity. These data demonstrate that pI:C-mediated protection from GVH-induced changes is due to increased rejection of grafted B6 parental cells by F1 NK cells, a phenomenon very similar, if not identical, to HR to bone marrow grafts.

Animals↗

PubMed1986

The effects of polyinosinic: polycytidylic acid (pI:C) on the GVH reaction: immunopathological observations.

The effects of polyinosinic: polycytidylic acid (pI:C) on the graft-versus-host (GVH) reaction were studied. The drug pI:C rapidly and markedly induces interferon and augments natural killer (NK) cell activity. GVH reactions were induced by injecting parental lymphoid cells intravenously into F1 hybrid mice. The development of a GVH reaction was monitored by measuring the plaque-forming cell (PFC) response to sheep red blood cells (SRBC) and by histological examination. When 30 X 10(6) B6 lymphoid cells were injected into B6AF1 mice, the recipients developed profound immunosuppression by 10-12 days post-GVH induction. In addition, pathological changes indicative of GVH reactions were seen in the spleen, lymph nodes, thymus, liver, lung, pancreas, and salivary gland of these mice. However, the treatment of B6AF1 recipients with pI:C prior to parental cell transfer markedly reduced the degree of suppression of the immune response, as measured by the PFC response to SRBC. Also, such mice failed to demonstrate the histological lesions of GVH disease. Treatment of donor mice with pI:C had no effect in preventing either GVH-induced immunosuppression or pathological changes. This study suggests that a pI:C-induced mechanism, possibly involving NK cells, is capable of regulating the GVH reaction.

Animals↗

PubMed1986

The graft-versus-host reaction and immune function. III. Functional pre-T cells in the bone marrow of graft-versus-host-reactive mice displaying T cell immunodeficiency.

Studies were performed to determine whether pre-T cells develop normally in the bone marrow of mice displaying thymic dysplasia and T cell immunodeficiency as a consequence of a graft-versus-host (GVH) reaction. GVH reactions were induced in CBAxAF1 mice by the injection of A strain lymphoid cells. To test for the presence of pre-T cells in GVH-reactive mice, bone marrow from GVH-reactive mice (GVHBM) was injected into irradiated syngeneic F1 mice and 30-40 days later thymic morphology and function were studied. Morphology studies showed nearly normal thymic architectural restoration; moreover, such glands contained normal numbers of Thy-1-positive cells. Functional pre-T cells were evaluated by transferring thymocytes from the irradiated GVHBM-reconstituted mice into T-cell-deprived mice. These thymocytes reconstituted allograft reactivity, T helper cell function and Con A and PHA mitogen responses of T-cell-deprived mice. These results suggest that the pre-T cell population in the bone marrow is not affected by the GVH reaction. Therefore, the T cell immunodeficiency associated with the GVH reaction is not due to a deficiency of pre-T cells in the bone marrow but is more likely associated with GVH-induced thymic dysplasia.

Animals↗

PubMed1986

The graft-versus-host reaction and immune function. IV. B cell functional defect associated with a depletion of splenic colony-forming units in marrow of graft-versus-host-reactive mice.

Studies were conducted to determine whether a functional B cell defect occurred in the bone marrow of mice experiencing a GVH reaction (GVHBM). GVH reactions were induced in AxCBA F1 adult mice by an injection of A strain lymphoid cells. The GVH reaction was confirmed by immunosuppression and thymus histology. At various intervals after GVH induction, GVHBM was tested for its ability to restore B cell function in adult thymectomized irradiated mice reconstituted with normal thymocytes. GVHBM cells obtained seven days after GVH induction restored but slightly the plaque forming cell (PFC) response to sheep erythrocytes and the mitogen response to lipopolysaccharide (LPS). GVHBM cells obtained 10 days or later failed to reconstitute the PFC or LPS responses. GVHBM cells suppressed neither the T or B cell function of normal spleen cells nor the LPS mitogen response of normal bone marrow cells. In addition, the splenic colony-forming units (CFU-s) in GVHBM were slightly decreased by day 10 after GVH induction and markedly depressed by day 22 after GVH induction. These results suggest that the GVH reaction may affect two different events in B cell differentiation. The early decrease in functional B cells that occurs before there is any change in the CFU-s population suggests a direct effect on B cell production, whereas the later absence of functional B cells could be due to the marked decline in stem cell production (CFU-s).

Animals↗

PubMed1985

Stimulation of the murine mixed leukocyte reaction: optimal proliferation of responders requires activation of stimulators.

In characterizing the functional properties of the stimulators in a mixed leukocyte reaction (MLR) we have established that the optimally effective stimulators must themselves be capable of undergoing the early stages of proliferative activation. We treated the stimulators with sublytic doses of a variety of agents (ouabain, ultra-violet radiation, 5-fluorouracil and colchicine), which have been reported to inhibit their stimulating capacity, and then in parallel experiments, we examined the treated cells for their capacity to stimulate allogeneic cells and their ability to respond to Con A. A direct correlation was found between the capacity of mitomycin C-treated splenocytes to stimulate allogeneic cells in an MLR and their ability to undergo changes characteristic of pre-S phase activation, namely, increases in K+ influx and in cell size, in response to Con A. This correlation also existed in the case of metabolically active but immunosuppressed splenocytes from mice undergoing late graft-versus-host (GVH) reaction: these cells did not show increase in potassium influx in response to Con A and their stimulating ability was inhibited. Using fluorescein labelled anti-Thy 1.2 as a marker for T cells in the stimulating population, it was shown that a considerable fraction (up to one third) of the blasts in a mixed leukocyte culture (MLC) originated in the mitomycin C-treated stimulating population. These data are consistent with the hypothesis that induction of maximal proliferation, in MLR, requires that certain of the cells (mainly T cells) of the stimulating population undergo the early stages of activation.

Animals↗

PubMed1985

Interleukin-1 and interleukin-2 defects associated with murine graft-versus-host-induced immunodeficiency.

In this study we investigated the mechanism, or mechanisms, involved in graft-versus-host (GVH)-induced T cell immunodeficiency. Chronic GVH reactions were induced in normal CBA X A F1 (BAF1) hybrid mice by the injection of parental A strain lymphoid cells. At various times (43-91 days) after GVH induction, the functional status of GVH T cells was assessed using interleukin-1 (IL-1) and interleukin-2 (IL-2) as probes. The response of GVH thymocytes to IL-1 was depressed when compared with normal thymocytes. Although GVH peanut-agglutinin-negative (PNA-) thymocytes did respond to IL-2 alone or IL-2 plus phytohemagglutinin (PHA), this response was significantly lower than the response of PNA- thymocytes from normal mice. In addition, GVH spleen cells failed to produce significant amounts of IL-2 when stimulated with concanavalin A. These results suggest that the long term immunosuppression associated with murine chronic GVH disease is due, at least in part, to a decrease in the responsiveness to IL-1 and IL-2, and to a marked deficiency in IL-2 production.

Animals↗

PubMed1985

The functional and histological basis for graft-versus-host-induced immunosuppression.

The GvH reaction resulting from the injection of parental strain cells into adult F1 hybrids suppresses both cell-mediated and humoral immune responses and is dependent on the donor-host combination and the number of parental cells used to induce the GvH reaction. The early suppression is due, at least in part, to the increased number of macrophages and the activation of suppressor macrophages which act directly on the T-helper cell and perhaps the B-cell as well. The macrophage suppression is associated with an increase in PGE production. The long-term T-cell immunodeficiency is mediated by GvH-induced thymic dysplasia resulting in a block or an arrest in T-cell differentiation and deficient IL-2 production. The B-cell immunodeficiency is associated with both a decrease in B-cell production from lymphoid progenitors and a decrease in CFU-s production. The GvH reaction induces 2 types of thymic lesion, a stress-related effect causing atrophy of the thymic cortex and a cytolytic process causing severe-to-moderate lesions in the thymic medulla as a consequence of injury to medullary epithelial cells and a loss of Hassall's corpuscles (thymic dysplasia). By employing the NK-cell-deficient beige mutation, it was shown that the severe-to-moderate thymic medullary lesions occurred in F1 mice only in those transplant situations in which the donor inoculum was of the +/bg genotype, regardless of the genotype of the recipient. It is proposed that activation of parental T cells may contribute to the early immunosuppressive events; however, the relatively permanent immunosuppression appears to be associated with NK-like effector cells which are capable of causing injury to lymphoid and epithelial tissue, especially epithelium of the thymic medulla. These studies raise the possibility that the GvH reaction may contribute to some T- and B-cell immunodeficiencies observed in the SCID and AIDS syndromes, as well as in patients following bone marrow transplantation.

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