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M J Dallman

Publications and source records attributed to M J Dallman.

At least 37 records · Page 2Linked to original sources
PubMed1997

The immune response following small bowel transplantation. II. A very early cytokine response in the gut-associated lymphoid tissue.

The small bowel has a unique amount of closely associated lymphoid tissue in the form of mesenteric lymph nodes (MLNs) and Peyer's patches (PPs). It is rather unclear how this may affect the immune response to transplants involving small bowel. It is clear, however, that host-derived leukocytes infiltrate this lymphoid tissue very rapidly after transplantation of small bowel, which suggests the possibility of an early immune response within this compartment. To investigate this possibility, we analyzed, using a semiquantitative reverse transcriptase-polymerase chain reaction, the level of cytokine transcripts within isolated MLNs and PPs for the first 7 days after small bowel transplantation. Heterotopic small bowel (n=32) transplants were performed using the following rat strain combinations: syngeneic Lewis (Lew)-->Lew (n=8), blood group D Agouti (DA)-->DA (n=8), allogeneic Lew-->DA (n=8), and allogeneic DA-->Lew (n=8). Two rats from each group were killed at 1, 3, 5, and 7 days after transplantation. RNA was prepared separately from PPs and MLNs before analysis of transcripts for interleukin (IL) 2, IL-4, IL-10, IL-6, IL-1alpha, and interferon (IFN) gamma. No increase in transcripts for IL-2 or IL-10 was observed in either PPs or MLNs of syngeneic grafts. A small rise in IL-6, IL-1alpha, and IFN-gamma transcripts was seen in MLNs and IFN-gamma transcripts in PPs of syngeneic grafts. In contrast, in allografts an extremely early increase in cytokine transcripts was observed; all cytokine transcripts tested were elevated within the first 24 hr after transplantation. Indeed, the peak response of both IL-2 and IL-10 occurred within 1 to 3 days after grafting. This early immune response in the lymphoid tissue may not be controlled by immunosuppression delivered only at the time of transplantation, and therefore may be responsible for the difficulty in achieving adequate immunosuppression in small bowel transplantation.

Animals↗
PubMed1997

Effect of one-HLA-haplotype-matched and HLA-mismatched blood transfusions on recipient T lymphocyte allorepertoires.

BACKGROUND: Pretransplant blood transfusion has a well-known beneficial effect on posttransplant graft survival. Recently, it has been proposed that the clinical benefit of transfusion is due to HLA-DR antigen sharing between the blood donor(s) and the recipient. Immunological studies have suggested that this might result from a functional deletion of donor-reactive cytotoxic T lymphocytes. METHODS: We investigated frequencies of alloreactive lymphocyte precursors with cytotoxic or interleukin-2-producing helper function by limiting dilution analysis in 10 renal dialysis patients before and after transfusion with fresh, allogeneic whole blood. Five patients received blood transfusions from donors matched for one HLA haplotype (or one HLA-B-DR antigen) and the other five patients received blood from fully HLA-mismatched donors. RESULTS: Contrary to some previous reports, frequency analysis of cytotoxic T lymphocyte precursors revealed no significant differences between the two treatment groups in terms of development of blood donor-specific hyporesponsiveness after transfusion. Split-well analysis of cytotoxic T lymphocyte precursors reactive with single-mismatched HLA antigens demonstrated that the effects of transfusion on alloreactive specificity are complex and may vary depending on the particular antigens mismatched between the recipient and blood donor. Analysis of donor-specific helper T lymphocyte precursor frequencies revealed a significant decrease of interleukin-2-producing cells 3 months after transfusion in the total patient population. This effect was most prominent in the recipients of HLA-mismatched blood, but it also exhibited some degree of nonspecificity, as frequencies of third-party reactive helper T lymphocyte precursors were also significantly reduced. CONCLUSIONS: Our overall results suggest that the degree of HLA matching between blood donor and recipient does not greatly influence the effect of blood transfusion on the T lymphocyte allorepertoire. The apparent induced down-regulation of helper T lymphocyte activity may play a role in the reported immunosuppressive effects of allogeneic blood transfusion.

Adult↗
PubMed1997

Patterns of graft infiltration and cytokine gene expression during the first 10 days of kidney transplantation.

Understanding of the events preceding acute cellular rejection of kidney transplants would be useful in the development of immunosuppressive strategies to prevent rejection. Information about these events in humans has been scarce, because of the lack of early, serial, biopsy samples. We took daily fine needle aspirates from kidney allografts for the first 10 days after transplant. Samples were analyzed by morphological cytology of graft-infiltrating cells, and reverse transcriptase-polymerase chain reaction for detection of interleukin (IL)-2, IL-4, IL-6, IL-10, and gamma-interferon gene expression. During the first 4 days, all of the grafts developed a low-grade monocyte-rich mononuclear cell infiltrate, accompanied by IL-10 gene expression. Thereafter, the infiltrates either remained stable or intensified. Of the 13 grafts with dense infiltrates, seven developed graft dysfunction. The remaining six did not, despite significant interstitial infiltrates. Both rejecting and nonrejecting dense infiltrates were associated with a biphasic pattern of IL-2 and gamma-interferon gene expression, preceding and accompanying lymphocytic graft infiltration. Grafts that did not develop dense infiltrates had no detectable IL-2 or gamma-interferon gene expression and did not suffer cellular rejection during the study period. The development of both rejecting and nonrejecting infiltrates was strongly associated with DR mismatches between donor and recipient. IL-2 and gamma-interferon gene expression are necessary, but not sufficient, for the development of acute cellular rejection in the first 10 days of kidney transplantation, and are more closely associated with the period leading up to rejection than with the period of graft dysfunction.

Acute Disease↗
PubMed1997

Semi-quantitative polymerase chain reaction analysis of cytokine and cytokine receptor gene expression during thymic ontogeny.

Semi-quantitative, polymerase chain reaction (PCR) is used to uncover the patterns of cytokine transcription in the mouse thymus from day 14 to day 20 of gestation, a time period which includes many of the important events in thymic ontogeny. Interleukin 4 (IL-4), IL-7 and interferon gamma (IFN-gamma) mRNA is abundant from fetal day (Fd) 14-16, corresponding with the period of rapid proliferation of immature thymocytes in vivo. As the level of mRNA for these cytokines diminishes, the induction and increased expression of IL-3 and IL-2 occurs. The transcription of these cytokines correlates temporally with the period of proliferation-dependent phenotypic differentiation between Fd 16 and 20. The thymic epithelium (TE)-derived cytokines including IL-1alpha, IL-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) begin to be transcribed between Fd 14-15 and show peak mRNA abundance from Fd 16-20. IL-5, tumour necrosis factor alpha (TNF-alpha) and LT (lymphotoxin or TNF-beta) constitute a fourth group of cytokines, along with the IL-4 receptor (IL-4R), which are transcribed at an even level throughout the fetal period. The IL-2 receptor beta chain (IL-2Rbeta) and IL-10 show abundant mRNA from Fd 14-20 and have a peak level of mRNA content on Fd 16. Taken together, these studies uncover complex, overlapping patterns of cytokine gene expression. The mRNA abundance and pattern of expression of each cytokine or cytokine receptor may indicate the relative contribution that it makes to different stages of fetal thymic ontogeny.

Animals↗
PubMed1997

Treatment with an HLA-peptide and cyclosporine A prolongs rat small bowel allograft survival.

BACKGROUND: The ultimate treatment for severe short bowel syndrome is small bowel transplantation (SBT). Current immunosuppression for SBT is relatively ineffective and toxic. Peptides derived from residues 75-84 of the HLA-B7 molecule are immunomodulatory in vitro, and in rodents, when combined with subtherapeutic doses of cyclosporine (CsA), prolong cardiac and skin allograft survival without altering the recipient's rejection kinetics to third party allografts. We investigated the effects of HLA-B7 peptide fragment in a rat model of SBT. METHODS: Heterotopic allogeneic SBT was performed in Dagouti (RT1a) to Lewis (RT1l) high-responder rat strain combination. B7.75-84 (40 mg/kg/d) and subtherapeutic CsA (10 mg/kg/d) were administered alone, or in combination, by gavage to allograft recipients on days 0 to 4 after SBT. Recipient pretreatment with B7.75-84 on days -14, -12, -10, and -7 followed by subtherapeutic CsA on days 0 to 4 after SBT was also carried out. Graft rejection was determined by the presence of a palpable abdominal mass on daily examination or by loss of more than 10% initial body weight. RESULTS: Without immunosuppression allografts rejected at a median time of 6 days (range, 5 to 7; n = 7). This was not significantly altered with either CsA therapy alone (median 6 days; range, 6 to 7; n = 6) or B7.75-84 alone (median, 5 days; range, 5 to 6; n = 6). Recipient combination therapy with B7.75-84 and CsA after allografting significantly prolonged allograft survival (median, 11 days; range, 9 to 13; n = 9), as did recipient B7.75-84 pretransplant therapy (median, 10 days; range, 9 to 12; n = 6), when administered over a 2-week period before allografting. CONCLUSION: Post-SBT recipient treatment with B7.75-84 produced statistically significant improvement in allograft survival only after combination with subtherapeutic CsA. Recipient pre-SBT treatment with B7.75-84 alone however, resulted in statistically significant improvement in allograft survival in combination with post-SBT subtherapeutic CsA. These synergistic effects may be valuable in achieving improved SBT survival clinically and warrant further exploration.

Animals↗
PubMed1997

Ex vivo adenovirus-mediated gene transfer and immunomodulatory protein production in human cornea.

One attractive strategy to prevent or control allograft rejection is to genetically modify the donor tissue before transplantation. In this study, we have examined the feasibility of gene transfer to human corneal endothelium, using a number of recombinant adenovirus constructs. Ex vivo infection of human corneas with adenoviral vectors containing lacZ, under transcriptional control of either cytomegalovirus (CMV) or Rous sarcoma virus (RSV) promoters, provided high-level gene expression, which was largely restricted to endothelium. Expression of the reporter gene persisted at relatively high levels for up to 7 days, followed by a decline to indetectable levels by 28 days. RT-PCR analysis of lacZ transcription showed a similar picture with a short period (3-7 days) of RNA transcription after infection. In contrast, adenoviral DNA persisted for at least 56 days. Subsequently, we examined the expression of a potential therapeutic gene, CTLA-4 Ig fusion protein. Following infection of human corneas with adenoviral vectors encoding CTLA-4 Ig protein, high levels of the fusion protein were detected in corneal culture supernatants for up to 28 days. This protein was functionally active, as determined by binding to B7.1 (CD80)-expressing transfectants. This study suggests that genetic alteration of donor cornea before transplantation is a feasible approach for preventing or controlling allograft rejection. Similar gene-based strategies might also be feasible to prevent rejection of other transplanted tissues or organs.

Abatacept↗
PubMed1996

The immune response following small bowel transplantation: I. An unusual pattern of cytokine expression.

Acute cell mediated graft rejection is frequently associated with an immune response dominated by cytokines like IL-2 and IFNgamma. While small bowel grafts are rejected acutely, there is little information on the type of immune response generated following transplantation and, in particular, whether the cytokine profile resembles that seen during the rejection of other solid organ grafts. In this paper we compare the expression of cytokines in isolated gut tissue following experimental small bowel transplantation with that in heart grafts. Heterotopic small bowel (n=32) and cardiac (n=32) transplants were performed using the following rat strain combinations: syngeneic Lewis (Lew) > Lew (n=8), blood group D Agouti (DA) > Lew (n=8) and allogeneic Lew > DA (n=8), DA > Lew (n=8). Two rats from each group were sacrificed at 1, 3, 5, or 7 days after transplantation. RNA was prepared separately from gut wall, after removing the Peyer's patches (PPs) and mesenteric lymph nodes (MLNs) and from heart. Cytokine (IL-1alpha, IL-2, IL-4, IL-6, IL-10 and IFNgamma) transcripts were analyzed using semiquantitative RT-PCR. Most notably, transcripts of only a single cytokine, IFNgamma, became progressively elevated with time in the rejecting small bowel grafts. This is in marked contrast to the findings presented here for rat cardiac grafts in which transcripts of all cytokines tested show an increase with rejection. This significant and steady increase in IFNgamma expression occurred before there was any clinical or histological evidence of rejection. These data demonstrate that the mechanisms of rejection in small bowel and other solid organ grafts are likely to be different. Further, the unique rise in IFNgamma expression in the gut wall may be a valuable and early indicator of graft rejection.

Animals↗
PubMed1996

Primary alloreactive cytotoxic T-lymphocytes are not commonly restricted by self-HLA class I antigens.

To assess the role of HLA Class I molecules in the indirect presentation of alloantigen, we have investigated the fine specificity and MHC restriction of in vitro primary alloreactive cytotoxic T-lymphocytes (CTL), using limiting dilution analysis of CTL precursor frequencies in HLA-mismatched responder-stimulator pairs. By employing split-well analysis of limiting dilution (LD) microcultures and third-party target cells bearing a stimulatory HLA Class I antigen alone or in combination with a single responder HLA antigen, we demonstrate that self-Class I restriction of HLA-A- or HLA-B-specific CTL precursors is not a common feature of the primary in vitro alloresponse. Higher frequencies of alloantigen-specific CTL precursors in the presence of self-HLA antigens were only detected in 5 of 31 limiting dilution assays established from seven different responder-stimulator pairs. In two cases, the higher precursor frequencies could be explained on the basis of Class II-restricted presentation of Class I-derived antigenic peptide and are supported by flow cytometric analysis of HLA antigen expression on target cells. The remaining 3 assays of this type were suggestive of Class I restriction but revealed only marginally higher frequency estimates. All other LD assays revealed lower CTL precursor frequency estimates in the presence of self-HLA Class I antigens. A higher antigen-specific CTLp frequency was not detected when targets shared three HLA Class I antigens with the responder, demonstrating that we had not biased the responses by selecting single HLA antigen-sharing targets in the other assays. Analysis of reactivity against PHA blast targets at the single cell per well level demonstrated that CTL reactive only with the original stimulator comprised the majority of lytic reactions. Heteroclitic CTL (i.e., CTL that recognize single HLA targets only and not the original stimulator) formed only a small fraction of total reactivity. Our results confirm the role of Class II antigens in the indirect presentation of alloantigen in vitro but suggest that HLA Class I antigens play a limited role in this phenomenon.

Autoantigens↗
PubMed1996

Enumeration of human alloreactive helper T lymphocyte precursor frequencies by limiting dilution analysis of interleukin-2 production.

We describe a limiting dilution assay for the enumeration of alloreactive interleukin-2 (IL-2) producing helper T lymphocyte precursors (HTLp). In place of the commonly used CTLL cell line, we have employed concanvalin A (ConA) stimulated rat thymocytes as IL-2 responsive indicator cells in a proliferation assay to detect IL-2 levels in limiting dilution microculture supernatants. The proliferation of ConA stimulated thymocytes induced by either recombinant IL-2 or culture supernatants could be blocked by co-incubation with a monoclonal antibody against the rat IL-2 receptor alpha chain, demonstrating the specificity of the response. Our investigations of alloantigen-induced IL-2 production show that (i) a minimum stimulator cell irradiation dose of 50-60 Gy is required to prevent backstimulation of microcultures; (ii) frequencies of alloreactive HTLp are significantly associated with HLA-DR antigen matching between responder and stimulator; (iii) HTLp frequencies detected in assays using B lymphoblastoid cell line stimulators are significantly higher than in assays employing peripheral blood lymphocyte stimulators but possibly reflect a degree of non-specific activation; and (iv) allosensitized responders exhibit altered kinetics of IL-2 production which may permit discrimination between sensitized and naive individuals. Our results both confirm and extend previous reports concerning such features of the alloresponse in humans and demonstrate that ConA stimulated thymocytes are a suitable alternative to CTLL as IL-2 responsive indicator cells in limiting dilution assays for HTLp analysis.

Animals↗
PubMed1996

Molecular biology of the allograft response.

Intensive basic research in immunology has provided us with a detailed understanding of the molecular events and interactions of the immune response that would have seemed incredible only a decade ago. An in-depth knowledge of the factors that initiate and control the immune response is, however, just the first step in learning how to manipulate this complex process for human benefit. One goal that seems quite attainable over the near term is a significant improvement in our ability to alleviate many of the problems resulting from allograft rejection--with newer drugs that not only are more effective, but are also much less hazardous or debilitating to transplant patients. Clearly the goal for the long term is the induction of allograft tolerance, rather than simply the modification of rejection. Genetic techniques will continue to serve as important tools in the research necessary to explore this new territory, and they are even now showing tremendous potential as sources of useful and effective immunotherapies.

Graft Rejection↗
PubMed1995

Cellular distribution and costimulatory function of rat CD28. Regulated expression during thymocyte maturation and induction of cyclosporin A sensitivity of costimulated T cell responses by phorbol ester.

CD28 has been identified in man and mouse as a potent costimulatory receptor on T cells. We have generated a mAb, called JJ319, to rat CD28 and show that it is expressed on virtually all peripheral rat alpha beta and on most gamma delta T cells, and on about half of NK cells. In contrast to the mouse but as in humans, most immature CD4+8+TCRlow thymocytes express little or no CD28, whereas CD28 expression is high on TCRintermediate and TCRhigh cells. mAb JJ319 very effectively costimulates T cell proliferation and IL-2 secretion by resting rat T cells. In contrast to results obtained in mice and humans, phorbol ester did not synergize in T cell activation with CD28-specific mAb but even induced sensitivity to cyclosporin A in T cell cultures that were optimally costimulated by mAbs to the TCR and to CD28. This result points to a novel effect of protein kinase activation by phorbol ester on signal transduction by TCR plus CD28 costimulation which only becomes apparent if, as in the rat, the TCR-mediated signal cannot be replaced by phorbol ester.

Animals↗
PubMed1995

Enhancement and inhibition of cellular responsiveness mediated by the anti-leucocyte monoclonal antibody NDS 71.

In this paper we describe the effects of a novel monoclonal antibody (mAb), which either inhibited or enhanced IL-2-stimulated proliferation of human peripheral blood leukocytes (PBL) at the same antibody concentration, under different culture conditions. An effect of this nature on IL-2-stimulated proliferation has not, to our knowledge, previously been described. The mAb NDS 71 bound to more than 98% of both freshly isolated and PHA-stimulated human PBL, with the level of binding markedly up-regulated on activation. An antigen of molecular weight 38-46 kDa was immunoprecipitated. When added to PBL at the beginning of in vitro culture, NDS 71 inhibited IL-2-stimulated proliferation. PHA-stimulated proliferation was enhanced by NDS 71 in identical conditions, demonstrating that the effects of NDS 71 on IL-2-stimulated proliferation were not merely due to non-specific inhibition of cellular proliferation. When NDS 71 was added to IL-2-stimulated PBL 4 h after the start of in vitro culture, proliferation was markedly enhanced, in contrast to the inhibition seen when the same concentration of antibody was added at the start of culture. In summary, NDS 71 markedly affected IL-2-stimulated proliferation, the same concentration of antibody causing either marked inhibition or enhancement, depending on the culture conditions.

Adjuvants, Immunologic↗
PubMed1995

Cytokines and transplantation: Th1/Th2 regulation of the immune response to solid organ transplants in the adult.

It has been very tempting to accept the suggestion that the route to rejection or tolerance of organ transplants is determined by T-helper type 1 and type 2 cells, respectively. Much of the data used to support this idea, however, is indirect and therefore cannot be used to imply a causal role for either population as suggested. Recent experiments have been aimed at further expanding knowledge in this area and conclude that the expansion of neither population alone inevitably results in graft damage or tolerance.

Animals↗
PubMed1995

Transplantation immunology.

The success of transplantation is such that it is now the treatment of choice for many of those requiring renal replacement therapy. The use of other solid organs, including liver, pancreas, heart and lung, continues to progress. This article reviews some recent advances in our understanding of the immunological response to alloantigen and xenoantigen.

Animals↗
PubMed1995

Spontaneous acceptance of rat liver allografts is associated with an early downregulation of intragraft interleukin-4 messenger RNA expression.

Liver allografts are not rejected in the fully incompatible Lewis-RT1(1) (LEW) to blood group D Agouti-RT1a (DA) rat strain combination despite an early infiltration by recipient mononucleated cells that initially display a phenotype, an ability to respond to interleukin-2 (IL-2) and donor-specific cytotoxicity indistinguishable from that observed in the rejected, DA to LEW combination. To further analyze the mechanism of this tolerance, we have compared in these two combinations, as well as in syngeneic grafts and in normal livers, the presence of intrahepatic cytokine transcripts (IL-1 alpha, IL-2, IL-4, IL-6, IL-10, tumor necrosis factor [TNF]-alpha, TNF-beta, and transforming growth factor [TGF]-beta) by a semiquantitative polymerase chain reaction (PCR) or by northern-blotting. In normal livers or syngeneic grafts, IL-1 alpha, TNF-beta, and TGF-beta were the only cytokines detected by these methods. The levels of all cytokine transcripts were increased in allogeneic grafts. Expression of cytokine transcripts was very similar in the two allogeneic strain combinations except IL-4, which was expressed at a much lower level in the nonrejected strain than in the rejected strain from day 2 onward. We conclude that selective downregulation of IL-4 gene expression is associated with, and a potential mediator of, the induction of tolerance in this model.

Acute-Phase Reaction↗
PubMed1994

Spontaneous acceptance of liver allografts in the rat. Analysis of the immune response.

In a model of arterialized rat liver transplantation, the biological indicators of liver dysfunction and the phenotype and in vitro function of graft-infiltrating cells have been compared in rejected (DA-to-Lew) and spontaneously accepted (Lew-to-DA) grafts, 2-8 days after grafting. Recipients of rejected and nonrejected allografts had, during this time, similar loss in body weight and plasma levels of transaminase. The markers of cholestasis, however, increased from days 3 and 4 onward in the recipients of rejected grafts, but remained low and similar in the recipients of nonrejected allografts and those of syngeneic grafts. From days 2 to 6 the phenotype, IL-2 responsiveness, and donor-specific cytotoxic potential of the leukocytes infiltrating rejected and nonrejected allografts were comparable. On days 7 and 8, although the proportion of T cell subpopulations was identical in both combinations, activated CD4+ graft-infiltrating cells were reduced in the nonrejected grafts. Also at this time, donor-specific cytotoxic cells were no longer detected in nonrejected grafts, whereas activity had reached a peak in the rejected grafts. These results suggest that liver grafts in both the LEW-->DA (grafts not rejected) and DA-->LEW (grafts rejected) strain combinations undergo tissue damage, but that the type of damage differs between the two combinations. Specifically, cholestasis was only observed in grafts that would subsequently be rejected. The difference in graft damage occurred at a very early time point (3 or 4 days after grafting), at which time neither the intensity nor phenotype of the graft infiltrate, its IL-2 responsiveness, or its cytotoxic potential varied between the two combinations. Thus, a lack of immune reactivity, as assessed by these parameters, does appear not to be responsible for spontaneous acceptance of liver transplants in the LEW-->DA strain combination.

Animals↗