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Biomedical subjects

Y Mullen

Publications and source records attributed to Y Mullen.

At least 91 records · Page 5Linked to original sources
PubMed1990

Fetal pancreas transplantation in miniature swine. IV. Suppression of DTH and MLR responses by treatment with ultraviolet light-irradiated peripheral blood lymphocytes.

Irradiation of peripheral blood lymphocytes of miniature swine with ultraviolet light prevented them from initiating proliferative responses in allogeneic mixed lymphocyte reactions and also reduced IL-2 production in these MLRs. When pigs were injected in a series of 4-5 weekly transfusions with UV-irradiated allogeneic PBL differing at the MHC, PBL of recipient pigs progressively responded less strongly to donor PBL in MLRs over the treatment period. These pigs also gave negligible delayed-type hypersensitivity responses to donor PBL at the end of the treatment period. Of the seven UV-irradiated PBL-treated pigs, four produced no antidonor PBL antibody and three produced antibody. Serum from the three antibody-producing pigs also suppressed MLRs of unrelated PBL. By contrast, pigs that received a series of injections of untreated allogeneic PBL gave strong DTH responses to donor PBL and heightened proliferation in MLRs with donor PBL, and all produced antidonor PBL antibody.

Animals↗
PubMed1989

Islet transplantation for the cure of diabetes.

This symposium format was similar to that used in the Kroc Foundation Meetings that Josiah Brown organized previously. As it had been at the Kroc Meetings, this memorial symposium dedicated to Dr. Brown was very successful in terms of the quality of the presentations, updating the most critical findings in islet transplantation and fostering the free exchange of provocative opinions. All participants well recognized that there has been tremendous advancement in islet isolation techniques, understanding of the immunological mechanisms underlying islet rejection, and the development of methods of immunomodulation to overcome rejection-related problems. Progress in these areas is one more step toward the final goal of providing new treatment for diabetics.

Animals↗
PubMed1989

DR antigen expression on vascular endothelium and duct epithelium in fresh or cultured human fetal pancreata in the presence of gamma-interferon.

The cells expressing MHC class II antigens play an important role in allograft rejection. We examined the expression of HLA DR antigens in human fetal pancreata ranging in gestational ages from 8 to 24 weeks. DR antigens were detected by immunohistochemical techniques using H4 and 40D MoAbs with immunoperoxidase staining and the ABC procedure. Vascular endothelium was identified by goat antibodies directed to human factor VIII-related antigens. DR expression on endothelium was further confirmed by double staining of tissue sections with H4 and anti-FVIII antibodies. In pancreata younger than 18 weeks of gestation, DR antigens were found on single cells that were randomly distributed throughout the pancreatic parenchyma. These cells resembled dendritic cells in morphology, as reported previously. Some vascular endothelial cells located in the intralobular connective tissues expressed DR antigens, but those in the pancreatic parenchyma were DR-negative. In 18-22 weeks, some endothelia of small vessels in the parenchyma became DR-positive. By 24 weeks, DR antigens were also expressed on medium-sized blood vessels, whereas intraislet capillaries and duct epithelia were DR-negative. The number of DR-positive cells increased rapidly from 11.2 cells/field (x400) in 12-14 week pancreata to 42.8 cells/field in 18-22 week pancreata. Most DR-positive cells in pancreata earlier than 18 weeks were dendritic-like cells, while those in older pancreata were endothelial cells. When pancreatic tissue fragments were cultured for two days in the presence of rIFN-gamma, vascular endothelium and duct epithelium, both of which were otherwise DR-negative, had become DR-positive. Even with this increase, DR-positive cell numbers were fewer in pancreata younger than 18 weeks of gestation as compared with those in older pancreata. We also detected clusters of epithelial-like cells that were clearly stained for DR antigens. These cells were located in the parenchyma where insulin positive cells were generally found. Their DR-antigens had not changed during the 3-day rIFN-gamma culture.

Dendritic Cells↗
PubMed1989

Fetal pancreas transplantation in miniature swine. II. Survival of fetal pig pancreas allografts cultured at room temperature.

Islet-allograft survival has been shown to be markedly prolonged in rodents when donor tissue has been precultured at 24 degrees C. In this study, the feasibility of this approach was tested in NIH minipigs transplanted with fetal pancreases. Collagenase-digested fetal pig pancreatic tissues survived in culture at 24 degrees C for 6-7 days and continued to grow in vitro at 37 degrees C after being transferred. These tissues no longer stimulated allogeneic lymphocytes in vitro, although some tissues cultured at 37 degrees C did. This allogeneic stimulation did not correlate to the number of major histocompatibility complex (MHC) class II-positive cells in stimulator pancreatic cultures. When transplanted into an omentum pouch of normal, nonimmunosuppressed minipigs, fresh fetal pancreatic tissues were rejected within 14 days. Tissues cultured at 24 degrees C grew, and beta-cells proliferated in minipigs treated daily with cyclosporin A (CsA) and azathioprine. Twelve normal minipigs were transplanted with 24 degrees C-cultured fetal pancreases: 8 pigs received no treatment, 2 received 14 mg.kg-1.day-1 CsA for 14 days, and 2 received 6-7 intravenous injections of platelets prepared from pooled farm-pig blood before grafting. Strong lymphocytic infiltration was detected in all grafts removed between 30 and 90 days posttransplantation. However, beta-cells were found on day 45 in one of five minipig pancreas grafts incompatible at the MHC loci and on days 60-90 in all three grafts compatible at MHC but incompatible at minor histocompatibility loci. Short-term CsA treatment did not prolong survival of allografts from farm pigs into minipigs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Expression of genetically determined diabetes and insulitis in the nonobese diabetic (NOD) mouse at the level of bone marrow-derived cells. Transfer of diabetes and insulitis to nondiabetic (NOD X B10) F1 mice with bone marrow cells from NOD mice.

The development of autoimmune diabetes in the nonobese diabetic (NOD) mouse is controlled by at least three recessive loci, including one linked to the MHC. To determine whether any of these genetic loci exert their effects via the immune system, radiation bone marrow chimeras were constructed in which (NOD X B10)F1-irradiated recipients were reconstituted with NOD bone marrow cells. Unmanipulated (NOD X B10)F1 mice, or irradiated F1 mice reconstituted with F1 or B10 bone marrow, did not display insulitis or diabetes. In contrast, insulitis was observed in a majority of the NOD----F1 chimeras and diabetes developed in 21% of the mice. These data demonstrate that expression of the diabetic phenotype in the NOD mouse is dependent on NOD-derived hematopoietic stem cells. Diabetogenic genes in the NOD mouse do not appear to function at the level of the insulin-producing beta cells since NOD----F1 chimeras not only developed insulitis and diabetes but also rejected beta cells within pancreas transplants from newborn B10 mice. These data suggest that the beta cells of the NOD mouse do not express a unique antigenic determinant that is the target of the autoimmune response.

Animals↗
PubMed1988

The effect of H-2 compatibility on pancreatic beta cell survival in the nonobese diabetic mouse.

The effect of major histocompatibility (MHC) antigens on the survival of newborn pancreatic iso and allografts was assessed in the nonobese diabetic (NOD) mouse, which develops a spontaneous autoimmune diabetes. The NOD mouse (H-2Kd,Db) rejects skin allografts from CBA (H-2k) within a 12-day period, indicating normal immune function toward alloantigens. A pancreatic allograft into the NOD mouse represents a presumed first-set allogeneic response, as well as a possible second-set immune response to islets. To assess the effect of donor H-2 antigen and the influence of autoimmune disease on pancreatic graft survival, newborn pancreata from various strains of mice were transplanted into diabetic NOD mice treated with 40 mg/kg/day cyclosporine (CsA) that prevented skin allograft rejection. The grafts were then harvested at day 10 to histologically assess the graft viability. CBA pancreatic grafts, incompatible at all MHC loci, showed the least lymphocytic infiltration, and good donor beta cell survival. Furthermore, CBA newborn pancreata under appropriate conditions were able to cure or improve the diabetic condition in 3/6 NOD mice. In the graft sharing class I MHC antigens, lymphocytic infiltration was significantly increased, while the donor beta cell number clearly decreased. The intensity of the graft destruction was intermediate in C57BL/6 allografts sharing H-2Db antigen, and strongest in BALB/c allografts sharing H-2Kd and in NOD isografts. The results indicate that in diabetic NOD mice the CsA dose controlling allograft rejection is incapable of controlling antiislet immunity. This antiislet immunity appears to exert its effect in an H-2-restricted manner. These findings may have important implications for the transplantation of pancreatic tissue in treating type I diabetes in humans.

Animals↗
PubMed1988

Influence of B-cell impairment on pancreatic acini in NOD mice and streptozotocin-induced diabetic rats.

Exocrine pancreatic function insufficiency, even of short duration, has been reported in juvenile-onset insulin dependent diabetic patients. To evaluate the status of pancreatic acini under decreased B-cell function, tissue insulin, amylase, chymotrypsinogen and trypsinogen in the pancreas were measured in streptozotocin-induced diabetic rats and non-obese diabetic mice in various conditions. In streptozotocin diabetic rats, a dissociation of three enzyme contents was demonstrated in the condition with discontinuation of insulin injection, i.e., a marked decrease in amylase, a significant increase in chymotrypsinogen, but no significant change in trypsinogen. This dissociation was markedly improved in the insulin-treated condition. In non-obese diabetic mice, these enzyme contents were not significantly changed although severe insulitis together with the marked decrease in insulin content was observed. These data show that the cessation of B-cell function alone does not cause insufficiency of exocrine pancreas.

Amylases↗
PubMed1987

Genetic control of diabetes and insulitis in the nonobese diabetic (NOD) mouse.

Genetic analysis of the development of diabetes and insulitis has been performed in the nonobese diabetic (NOD) mouse strain, a model of insulin-dependent (type I) diabetes mellitus. (NOD X C57BL/10)F1, F2, and (F1 X NOD) first-, second-, and third-backcross generations were studied. The data obtained were consistent with the hypothesis that diabetes is controlled by at least three functionally recessive diabetogenic genes, or gene complexes, one of which is linked to the MHC of the NOD. In contrast, pancreatic inflammation leading to insulitis was found to be controlled by a single incompletely dominant gene. One of the two diabetogenic loci that is not linked to the MHC appears to be essential for the development of severe insulitis. This diabetogenic gene may be identical to the gene that controls the initiation of the autoimmune response that progresses to insulitis. Although this gene appears to be functionally recessive in its control of diabetes, it is incompletely dominant in its control of insulitis. The MHC-linked diabetogenic gene, although not required for the development of insulitis, apparently influences the progression of the autoimmune response since NOD MHC homozygotes in the backcross generations displayed the highest incidence and most severe cases of insulitis. Interestingly, we have found two MHC heterozygous backcross females that have become diabetic, suggesting that either the MHC-linked diabetogenic gene is not strictly recessive or that a recombination event has occurred between the diabetogenic gene and the K or I-A regions of the MHC. The third diabetogenic locus appears to influence the progression of severe insulitis to overt diabetes. In animals homozygous at this locus, diabetes may result from a decreased ability to develop a protective suppressor response to the autoimmune process.

Animals↗
PubMed1987

An improved cryopreservation procedure for human fetal pancreas tissues.

Improved viability and function of insulin-producing beta (B) cells of frozen-stored human fetal pancreatic tissue was obtained by a two-step method utilizing high concentrations of dimethyl sulfoxide (DMSO). Human fetal pancreata (14-23-week gestation) obtained from pathologic abortions were teased and cultured overnight. Prior to freezing the tissues were immersed in 0.9% saline containing 0.5 M DMSO for 30 min (room temperature) and then placed in 2.1 M DMSO on ice for 5 min. The tissues were frozen by the method previously developed in our laboratory and stored at -196 degrees C. The frozen-stored tissues were subsequently thawed at 24 degrees C and cultured overnight before viability testing. Viability and function of the B cells were assessed by several specific assay methods; glucose plus theophylline-induced insulin release during static incubation and perifusion, 3H-leucine incorporation into insulin, and insulin content of the tissue grown in athymic mice for 7 days. The response to glucose plus theophylline stimulation, measured on the frozen-thawed tissue one day after thawing, was 80% of the level measured in control tissue maintained in organ culture. Frozen-thawed tissues maintained in organ culture for 1 week responded comparably in the in vitro assay systems. The insulin content of frozen-thawed pancreatic tissue removed from athymic mice 1 week after transplantation was approximately 60% of the amount measured in the control grafts. These results demonstrate the utility of our procedure in the maintenance of the viability and function of frozen-stored human B cells both in culture and after transplantation.

Animals↗