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T Oberley

Publications and source records attributed to T Oberley.

11 recordsLinked to original sources
PubMed2000

Clonotype analysis of human alloreactive T cells: a novel approach to studying peripheral tolerance in a transplant recipient.

The recognition of allo-MHC and associated peptides on the surface of graft-derived APC by host T cells (direct pathway allorecognition) plays an important role in acute rejection after organ transplantation. However, the status of the direct pathway T cells in stable long term transplants remains unclear. To detect alloreactive T cell clones in PBL and the allograft during the transplant tolerance, we utilized RT-PCR instead of functional assays, which tend to underestimate their in vivo frequencies. We established alloreactive CD4+ and CD8+ T cell clones from peripheral blood sampled during the stable tolerance phase of a patient whose graft maintained good function for 9 years, 7 without immunosuppression. We analyzed the sequence of TCR Vbeta and Valpha genes and made clonotype-specific probes that allowed us to detect each clone in peripheral blood or biopsy specimens obtained during a 1-year period before and after the rapid onset of chronic rejection. We found an unexpectedly high level of donor HLA-specific T cell clonotype mRNA in peripheral blood during the late tolerance phase. Strong signals for two CD4+ clonotypes were detected in association with focal T cell infiltrates in the biopsy. Chronic rejection was associated with a reduction in direct pathway T cell clonotype mRNA in peripheral blood and the graft. Our data are inconsistent with the hypothesis that direct pathway T cells are involved only in early acute rejection events and suggest the possibility that some such T cells may contribute to the maintenance of peripheral tolerance to an allograft.

Amino Acid Sequence↗
PubMed1998

Analysis of primate renal allografts after T-cell depletion with anti-CD3-CRM9.

BACKGROUND: FN18-CRM9 is a CD3-specific immunotoxin that is capable of depleting CD3+ T cells. Pretreatment of rhesus monkeys with this agent before transplantation can induce donor-specific tolerance and "split tolerance" to renal allografts. METHODS: Heterotopic renal transplants were performed on monkeys that received posttransplant FN18-CRM9. Histological and immunohistological staining, as well as analysis of the intragraft cytokine profile by reverse transcriptase polymerase chain reaction, was performed on percutaneous allograft biopsies. RESULTS: Experimental monkeys had significant prolongation of allograft survival. Although an interstitial, mononuclear cell infiltrate was seen in all of the renal transplants, there was minimal evidence of acute cellular rejection. Histological evidence of alloantibody-mediated damage was detected 3 to 5 months after transplantation in the monkeys treated with FN18-CRM9. Immunohistology demonstrated the reappearance of CD3+ and CD4+ T cells, as well as CD20+ B cells, in the grafts. Cytokine analysis demonstrated expression of interferon-gamma. An intact anti-donor IgG response was seen. CONCLUSION: Treatment of monkeys with FN18-CRM9 immediately after transplantation significantly prolongs renal allograft survival. Allograft biopsies demonstrate a lack of acute cellular rejection; however, alloantibody-mediated graft damage and rejection occur, with an intact anti-donor IgG response. The intragraft expression of the interferon-gamma may reflect this ongoing humoral rejection. These data suggest that even a brief period of T-cell allosensitization may lead to humorally mediated allograft damage. Efforts to achieve tolerance with posttransplant FN18-CRM9 will require modification of the protocol to deplete T cells before allosensitization exposure or to supplement the posttransplant immunomodification strategy.

Animals↗
PubMed1998

Manganese superoxide dismutase overexpression attenuates MPTP toxicity.

There is substantial evidence implicating mitochondrial dysfunction and free radical generation in the neurotoxicity of MPTP. Manganese superoxide dismutase (MnSOD) is the primary antioxidant enzyme protecting against superoxide radicals produced within mitochondria. Overexpression of human MnSOD in transgenic mice resulted in increased MnSOD localized to mitochondria in neurons and a 50% increase in total MnSOD activity in brain homogenates. We found that MPTP toxicity was significantly attenuated in the MnSOD transgenic mice which overexpress the human manganese superoxide dismutase gene, with these mice showing threefold greater dopamine levels than controls following MPTP. There were no alterations in MPP+ levels, suggesting that the effects were not due to altered metabolism of MPTP. A significant increase in 3-nitrotyrosine levels was seen in littermate controls but not in transgenic mice overexpressing human MnSOD. These results provide further evidence implicating mitochondrial dysfunction and oxidative damage in the pathogenesis of MPTP neurotoxicity.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed1998

Primate renal transplants using immunotoxin.

BACKGROUND: T-lymphocyte depletion 7 days before transplantation with immunotoxin FN 18-CRM9 has resulted in tolerance to subsequent renal allografts. We tested the effect of giving immunotoxin on the day of the transplantation and evaluated its effect on rhesus monkey and allograft survival, on antibody production, and on T-cell recovery. METHODS: Major histocompatibility complex mismatched renal allografts were performed in rhesus monkeys. Immunotoxin was given starting on the day of transplantation, with and without prednisone and mycophenolate mofetil for 3 days. T-cell subsets and alloantibody levels were measured by flow cytometry. The ability of treated monkeys to develop antibody to tetanus, diphtheria, and xenoantibody was measured. Histology of renal transplants was read in a blinded manner. RESULTS: Immunotoxin started on the day of transplantation resulted in prolonged allograft survival in all treatment groups. Graft loss between days 50 and 135 was most often due to interstitial nephritis. Later graft loss was due to chronic rejection. Monkeys had intact antibody responses to alloantigen, tetanus, diphtheria, and xenoantibody. Their CD4 cells recovered gradually over 6 months. CONCLUSIONS: Immunotoxin reliably prolongs renal allograft survival when started on the day of transplantation, but interstitial nephritis and chronic rejection limit the development of long-term tolerance. T-cell-dependent B-cell responses remain intact after treatment.

Animals↗
PubMed1997

Extended culturing of androgen-responsive human primary epithelial prostate cell isolates by continuous treatment with interstitial collagenase.

Continuous culturing of two distinct human prostate specimens in the presence of interstitial collagenase added directly to conventional medium resulted in the isolation and extended growth of primary epithelial prostate cell (PEPC) cultures from each. Both continued to proliferate substantially beyond the average time determined for analogous untreated epithelial prostate isolates. Both repeatedly stain positive for keratin and are characteristically epithelial in morphological appearance and growth model. Both express androgen receptor mRNA and stain positive for androgen receptors. PEPC-2 displays an androgen dose-dependent stimulation of cell proliferation, as well as specifically binding 3H-R1881. PEPC-1 exhibits a hypotetraploid karyotype with loss of the Y chromosome. PEPC-2 conserves a normal human ploidy, including the Y chromosome, although there is extensive random chromosome loss. Elimination of the collagenase from the medium resulted in decreased cellular proliferation and accumulation of stainable collagen in both PEPC cultures. Neither PEPC isolate produced tumors in male nude mice, whether injected alone, mixed with matrigel, or combined with prostate or bone fibroblastic cells.

Aged↗
PubMed1992

Sonography of renal transplants in dogs: the effect of acute tubular necrosis, cyclosporine nephrotoxicity, and acute rejection on resistive index and renal length.

Results of studies on the accuracy of the resistive index as a predictor of acute renal transplant rejection have varied widely. Clinical evaluations are limited by the inability to control the numerous coincidental factors that affect vascular resistance. We performed a controlled study in dogs to isolate the effects of acute tubular necrosis, cyclosporine toxicity, and acute rejection on the resistive index, and to compare them with a population of normal control subjects. By doing so, we hoped to identify the patterns of change in the resistive index over time and possibly explain the wide spectrum of resistive index data reported in the literature. Resistive index, a parameter calculated from relative systolic and diastolic velocity, indicates parenchymal resistance to perfusion. Since an increase in renal length also has been reported useful in predicting rejection, we studied changes in length in each of the isolated conditions. The normal control group (four dogs) had heterotopic autotransplantation with minimal cold ischemic time. The acute tubular necrosis group (six dogs) had heterotopic autotransplantation with 1 hr of warm ischemic time. The cyclosporine toxicity group (four dogs) was allowed approximately 3 months to heal from heterotopic autotransplantation. Very high (toxic) doses of cyclosporine were then administered. The acute rejection group (five dogs) had heterotopic allografting with minimal cold ischemic time. No medications were administered. In all groups, the abnormalities induced were confirmed by biopsy. Creatinine levels were also used to monitor cyclosporine toxicity. In the normal control and acute tubular necrosis groups, resistive index increased immediately after surgery, returning to baseline within 10 days. Renal length increased slightly in both groups, but the duration of increase was longer in the acute tubular necrosis group. No significant change in resistive index or renal length was seen in the cyclosporine toxicity group. In the acute rejection group, an initial decrease in resistive index during the mild to moderate phase was followed by a rapidly progressive increase with worsening rejection. Renal length increased progressively beginning immediately after surgery. Our study determined the patterns of change in resistance and renal length over time as caused by the isolated pathologic states. Our finding that vascular resistance decreased in mild to moderate acute rejection was unexpected, since almost all the literature reports resistive index elevation. This may explain some of the conflicting results obtained in Doppler investigations of rejection. Our results on renal length reinforce the positive clinical reports of its predictive value in rejection.

Animals↗
PubMed1991

Disruption of cellular energy balance by suramin in intact human prostatic carcinoma cells, a likely antiproliferative mechanism.

The antiparasitic drug, suramin, has antiproliferative effects in human carcinoma cells. It has been suggested that this occurs through blockade of growth factor-receptor interactions. Three types of evidence that suramin rapidly inhibits cellular respiration or disrupts cellular energy balance in intact cells of the human prostate carcinoma cell line, DU145, are presented. Beginning at approximately 10(-4) M, suramin rapidly causes dose-dependent inhibition of tetrazolium conversion by mitochondrial dehydrogenases in intact cells, demonstrating an inhibition of respiration. This effect is reversed by exchange with suramin-free media but not by pretreatment with serum, epidermal growth factor, insulin-like growth factor I, acidic and basic fibroblast growth factors, or calcium. Rhodamine 123 (10 micrograms/ml) uptake by mitochondria in intact DU145 cells is inhibited in the presence of 10(-3) M suramin. Treatment with 10(-4)-10(-3) M suramin causes the loss of rhodamine 123 from cells with mitochondria prestained with rhodamine 123, indicating that suramin is acting as an ionophore or respiratory poison. Also shown by electron microscopy are progressive toxic changes in mitochondria of DU145 cells within 1 h after treatment with 10(-4) M suramin. These data indicate that in intact DU145 cells 10(-4) M suramin rapidly disrupts cellular energy balance or respiration as seen by three studies of mitochondrial state. Disruption of energy balance or respiration represents a likely antiproliferative mechanism, as is thought to be a primary mechanism for the action of suramin in parasitic diseases. This proposed mechanism of action for suramin can explain the most prominent observed clinical toxicities of nephrotoxicity, adrenal toxicity, coagulopathy, and demyelinating neuropathy.

Biological Transport↗
PubMed1986

Alterations in rat renal glucose transport following in vivo use of Freund's adjuvant.

Alterations in rat renal glucose transport following in vivo use of Freund's adjuvant were examined. Lewis-Brown Norway rats were placed in four separate injection groups: tubular basement membrane plus adjuvant [complete Freund's adjuvant (CFA) plus pertussis]; adjuvant (CFA plus pertussis); CFA only; or pertussis only. Renal handling of glucose was assessed 14 days after a single injection. No in vivo changes were noted. No histologic differences among groups were noted. However, brush border membrane vesicles prepared from animals in groups 1, 2, and 3 showed a marked decrease in glucose uptake. Further, Michaelis-Menten kinetics demonstrated a decrease in apparent Km and Vmax for glucose in groups 1, 2, and 3. CFA alone can cause a change in brush border membrane vesicle uptake of glucose. The pathogenic mechanism behind CFA-induced transport changes remains unclear. However, studies employing CFA cannot dismiss Freund's adjuvant as "inert" and must take into account functional changes created by CFA alone.

Animals↗