Biomedical subjects
Publications and source records attributed to R M Gorczynski.
LPS blasts inhibited graft rejection in multiple minor incompatible strain combinations but not in presensitized animals or in major histoincompatible combinations.
Endothelial monolayers were prepared from neonatal heart or liver tissue of Lewis (Le) rats. Cells in their first passage of culture were used to investigate the short-term (1 hr at 37 degrees) binding of 51Cr-labelled Le rat lymphocytes prepared from the mesenteric lymph node (MLN), peripheral lymph node (PLN) or Peyer's patches (PP) to those endothelia, or the activation by concanavalin A (Con A) or irradiated (Lewis x Brown Norway)F1 (LBNF1), of Le cells on the monolayers after 84 hr in culture. MLN and PP showed preferential binding to, and activation on, liver endothelium compared with heart endothelium (approximately twofold difference), while the converse was seen with PLN. No inhibition of binding was seen with antibodies to intracellular adhesion molecule-1 (ICAM-1) or lymphocyte function-associated antigen-1 (LFA-1). Preincubation of endothelial cells with plasma isolated from the portal or hepatic vein of normal adult mice (5% plasma, 37 degrees for 14 hr) caused a 1.5-2-fold stimulation of binding of MLN/PP to heart endothelium, which was inhibited (> or = 75%) by anti-ICAM-1 or anti-LFA-1, and a fourfold stimulation of binding to liver endothelium, which was not inhibited by these monoclonal antibodies (< or = 25% inhibition). In contrast, antibodies to tumour necrosis factor-alpha (TNF-alpha) or interleukin-6 (IL-6) caused inhibition of activation of liver endothelium (> or = 75%), while producing little affect on activation of heart endothelium. Similar results were seen when lymphocyte activation on endothelial cells rather than adhesion cells was investigated. Our data suggest a heterogeneity in lymphocyte-endothelial interactions, which is further regulated, under physiological conditions, by the liver.
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C3H/HEJ mice injected with irradiated multiple minor incompatible B10.BR lymphoid cells via the portal vein showed delayed rejection of subsequent B10.BR skin grafts. Similar delayed rejection was produced by lateral tail vein injection of B10.BR hepatic mononuclear cells or H-2k cells pulsed in vivo with B10 minor histocompatibility antigens. Inhibition of C3H anti-B10.BR immunity in vivo (assessed by delayed graft rejection) and in vitro (assessed by B10.BR-induced lymphokine production) can be transferred by radioresistant, plastic-adherent F4/80+33D1-CD4-CD8-alpha beta TcR-gamma delta TcR- mononuclear hepatic cells from (C3H/HEJ x C3H.SW)F1 mice injected 36 hr earlier with 100 x 10(6) irradiated spleen cells. By 10 days post-injection, cells transferring delayed rejection are radiosensitive, plastic non-adherent, F4/80-33D1-CD4-CD8- alpha beta Tc+- gamma delta TcR+ cells. Injection of interleukin-2 (IL-2) in vivo into mice receiving pretreatment with B10.BR cells via the portal vein, or adoptive transfer into such mice of immune anti-B10.BR lymphoid cells, abolished delayed rejection on subsequent skin grafting. Delayed rejection or modulation of lymphokine production was associated in all cases with suppression of IL-2 production and preferential retention of IL-4 production from cells stimulated in vitro.
Adult Lewis rats received syngeneic accessory small bowel transplants (SBT) with venous drainage to the portal vein (PV) or the inferior vena cava (IVC)/hetero-portal (HP) or hetero-systemic (HS) grafts, respectively. At varying times thereafter (5-300 days post-transplantation) animals were killed and cells from different lymphoid organs were tested in vitro for their generation of lymphoproliferative and cytotoxic T-cell responses, as well as their ability to produce a variety of lymphokines after alloantigen or mitogen stimulation. Despite marked decreases in cell recovery in Peyer's Patches of HS rats, no significant loss of cell function (on a per cell basis) was noted in any animal group. Total recovered activity per organ was decreased in small intestinal tissue (host and graft) of HS recipients regardless of the assay under study.
Adult rats received syngeneic accessory small bowel grafts with venous drainage to either the portal vein (hetero-portal) or the inferior vena cava (hetero-systemic). Lymphoid cell recovery in different lymphoid organs (spleen, pooled peripheral lymph nodes, mesenteric nodes, Peyer's Patches) was evaluated at varying times (days 0-300) post-grafting. While minimal changes were observed for cell recovery in other organ tissues, lymphocyte recovery in Peyer's Patches of both host and graft small intestine of hetero-systemic animals was decreased from 10- to 100-fold with respect to hetero-portal recipients or non-operated controls. These changes were seen throughout the time course of the study. In additional experiments, lymphoid cells from different organs/donors were labelled in vitro with 111In and injected intravenously into normal/transplanted recipients. Recovery of 111In in various organs was assessed at 1 and 6 h postinjection. The major change seen was in the decreased ability of mononuclear cells derived from Peyer's Patches to migrate to small intestinal tissue (host and graft) in hetero-systemic recipients. In addition, Peyer's Patch cells from these animals 'homed' poorly to small intestine in non-operated animals by comparison with cells from normal rats (or hetero-portal donors).
We examined the hypothesis that FK 506 would induce graft acceptance after lung transplantation. Left lung allotransplantation was performed in size-matched mongrel dogs allocated to control (no immunosuppression, n = 3) and FK 506 (n = 5) groups. FK 506 (1.2 mg/kg intramuscularly every day) was given on posttransplantation days 0, 1, and 2. No other immunosuppressive agents were administered to either group. Chest x-ray and transplant lung physiologic assessments were performed on the fifth day and weekly thereafter. On day 29 an open lung biopsy and a third-party skin graft were performed. Lymphocytes were harvested and frozen from the recipient peripheral blood before transplantation and on days 8 and 29 afterwards for assessment in mixed lymphocyte reaction. Dogs were killed when their chest x-ray films showed allograft opacification or when the skin graft was rejected. Control lungs were all rejected after a median of 5 days. In the FK 506 group, one of five dogs aspirated during the fifteenth-day assessment and was killed, on the twenty-ninth day, because of severe rejection. At day 29, in the other four dogs, the transplanted lung yielded an arterial oxygen tension of 613 +/- 25 mm Hg (mean +/- standard deviation) and lung biopsy specimens showed no abnormalities histologically. These four dogs rejected third-party skin grafts after a median of 10 days. In two FK 506 dogs, mixed lymphocyte reaction at day 8 showed suppression of proliferation responses against donor and third-party lymphocytes. By day 29 responses against third-party lymphocytes had returned to almost preoperative levels, whereas antidonor responses were still suppressed. After skin graft rejection and killing, one of four dogs showed no sign of rejection, and the other three showed minimal to mild lung rejection at the time they were killed. We conclude that a 3-day course of 1.2 mg/kg of FK 506 induced prolonged graft acceptance after lung transplantation in dogs.
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This study presents evidence for the conservation of Drosophila per gene homologs in mammalian DNA and for their expression in a number of tissues which are involved in various aspects of circadian timekeeping. Distinct 5 kb sequences, which hybridized to a non repetitive fragment of the Drosophila per gene under stringent conditions, were detected by Southern blotting. Sequences homologous to per gene of Drosophila were also amplified from rat and mouse brain cDNA libraries and from a mouse anterior hypothalamus and human hypothalamus libraries. Degenerate PCR primer design was based on conserved segments of the per protein. The per homologs were shown directly (by RT-PCR) to be expressed in hamster and mouse SCN, in hamster heart and in Aplysia and Bulla eyes.
Immunization of naive or specifically primed C3H/HEJ with irradiated B10.BR spleen cells via the hepatic portal vein leads to an antigen specific decrease in the proliferative and cytotoxic response to B10.BR antigen assayed in vitro (and to increased graft survival of B10.BR grafts in vivo). This effect seems to be mediated in the main by a decrease in IL-2 production from CD4+ T lymphocytes of mice given antigen by the portal route, which is in turn caused by a decreased precursor frequency of IL-2-producing cells. No clear decrease in IL-4 production was seen. Hepatic APC isolated from mice receiving antigen via the portal vein were unable to induce IL-2 production from a C3H/HEJ anti-B10.BR cell line in vitro, in contrast to splenic APC derived from the same mice. Even when antigen was given by conventional (systemic) intravenous routes (in this case via the lateral tail vein) hepatic APC isolated from those mice were unable to stimulate IL-2 production from this cell line. Furthermore, 24 h exposure of a cell line to antigen pulsed hepatic APC left those cells refractory to a subsequent restimulation with antigen presented by splenic APC. Spleen lymphoid cells from primed mice challenged in vivo with B10.BR liver cells (i.v.) were similarly unable to produce IL-2 on rechallenge in vitro with irradiated B10.BR spleen cells, though no defect was seen if in vivo challenge was with B10.BR spleen cells. These data imply that presentation of multiple minor cell surface antigens by hepatic APC leads to specific anergization of IL-2 producing T cells, in a fashion which seems to be distinct from that previously reported as due to 'veto-like' activity.
Pretransplant transfusion of multiple minor histoincompatible spleen cells to naive recipient mice by the portal vein suppresses the ability of those animals to reject skin grafts from mice syngeneic with those used for transfusion, and decreases in vitro immunity on rechallenge with the same antigens, by comparison with mice receiving transfusion by the lateral tail vein. We have shown elsewhere that this is correlated with a diminished activation of Th1 cells for IL-2 production, without apparently affecting activation of Th2 cells for IL-4 production. Similar data are obtained by merely infusing hepatic (vs. splenic) antigen-presenting cells (APC) into normal mice, or by challenging immune cells in vitro with antigen-pulsed hepatic (vs. splenic) APC. However, when antigen-pulsed splenic APC are incubated with immune T cells in the presence of anti-LFA-1 monoclonal antibody (Mab), selective activation of Th2 cells (as is seen with hepatic APC) again occurs at the expense of activation of Th1 cells. Anti-LFA-1 Mab causes little perturbation in lymphokine production from T cells stimulated with hepatic APC. Using cDNA probes for IL-2 and IL-4 we show that T-cell activation in the presence of anti-LFA-1 Mab leads to selective inhibition of transcription of IL-2 mRNA.
C3H/HEJ mice repeatedly exposed to rotational stress (45 rpm for 30 min) show a decreased immune response after challenge with sheep erythrocytes (SRBC) or multiple minor histoincompatible skin grafts (B10.BR) in association with the cues present when initially exposed to the physical stress. This conditioned immunosuppression is associated with diminished antibody production and enhanced skin graft survival. When previously conditioned mice were mated with normal (nonconditioned) males and reexposed to conditioning cues during pregnancy (days 13, 16, and 19 of gestation), the offspring of these mice were also found to produce a decreased antibody response (and decreased skin graft rejection response) when tested at 7 weeks of age. This altered immune response in the offspring occurred in the absence of any deliberate direct manipulation of these mice. In a further study a reciprocal crossover design was used to investigate whether altered immunity in the offspring was dependent upon the treatment of the biological mother and/or the fostering mother. The most marked immunosuppression was seen in offspring born to, and fostered on, conditioned mothers reexposed to cues previously associated with physical stress. However, even the offspring of of normal C3H/HEJ matings developed a reduced immune response if they were fostered on conditioned mothers reexposed to stress-associated cues. These data imply that the developing immune system is regulated by factors in the colostrum/feto-placental unit which are modified by conditioning phenomena.
With the accompanying paper these studies provide further evidence that functionally important lymphoid cell trafficking takes place in mesenteric tissue under the apparent control of factor(s) filtered by hepatic tissue. The role that these regulatory mechanisms have to play on survival of vascular allografts with venous drainage into either the portal or systemic circulation remains to be elucidated.
These data indicate that in rat heterotopic syngeneic SBT, the venous drainage system of the graft has profound effects on cell recovery in the gut-associated lymphoid tissue of both graft and host. Depending on the functional status of the same lymphoid tissue (Gorezynski, personal communication, 1992) one could thus anticipate significant perturbation of host-antigraft (and GVH) reactivity in allogeneic situations according to the venous drainage used. The mechanism(s) responsible for these effects have not been investigated. However, one testable hypothesis is that (a) factor(s) from the gut can control lymphoid recirculation within the mesenteric lymphoid tissue, and that this (these) factor(s) are absorbed under normal circumstances by hepatic tissue.
Naive CBA mice injected with AKR spleen cells via the portal vein (p.v.) subsequently showed decreased stimulation in vitro in a primary MLR or cell-mediated lympholysis assay with irradiated AKR stimulator cells. No inhibition of stimulation by B10.BR cells is seen. These mice also show specific prolongation of survival of AKR skin grafts in vivo and diminished capacity for in vivo priming for (secondary) anti-AKR responses in vitro. These effects are not seen if initial challenge is with AKR cells injected subcutaneously (s.c.) or via the lateral tail vein (i.v.). Moreover, if immune CBA anti-AKR mice are similarly challenged with AKR cells via the portal vein, no suppression of anti-AKR immunity is elicited, as determined by subsequent in vitro assays or in vivo graft rejection. However, spleen cells from CBA anti-AKR immune mice can be used to induce, in further naive CBA mice, a specific suppression of subsequent anti-AKR graft reactivity (assayed in vitro or in vivo). Active T cell-mediated suppression can be documented using both these protocols though the additional involvement of specific serum-mediated suppression cannot be eliminated.
Mice of different ages were subjected to repeated exposure to cyclophosphamide:saccharin (conditioned), or cyclophosphamide:saccharin followed by saccharin only (extinguished). Only young animals in the former group showed a decreased IgG antibody-forming-cell (AFC) response after challenge with sheep erythrocytes (SRBC) in the presence of saccharin. When irradiated conditioned young animals were used as recipients of antigen-challenged spleen cells from nonconditioned mice they were found to support a greater immune response than nonconditioned recipients. Similarly, cells from conditioned young mice gave a greater immune response in naive recipients than did cells from nonconditioned mice. Only when cells from conditioned young mice rather than conditioned "aged" mice were studied in irradiated conditioned young recipients was immunosuppression observed. These data are most consistent with a specific host cell:environment interaction being responsible for the conditioned immunosuppression observed in young mice. A deficit in both cells/environment apparently occurs during aging. At least one of these deficiencies seems related to loss of a suppressor T-cell population with age in conditioned mice.