摘要:

The area under the blood concentration vs. time curves of cyclosporine (24 hr) were determined nonspecifically by the polyclonal RIA, specifically by a monoclonal RIA and by HPLC after an oral and an i.v. dose of CsA in 10 renal transplant patients. The mean blood concentrations determined by monoclonal RIA were 10–20% higher than those measured by HPLC, whereas the concentrations assessed by polyclonal RIA were >100% higher than those determined by HPLC. As a corollary, the pharmacokinetic parameters (clearance, volume of distribution, and systemic availability) differed when the results from the 3 methods were compared. The RIA/ HPLC concentration ratio of CsA was higher after oral than after i.v. dosing when RIA measurements were performed by the polyclonal but not by the monoclonal RIA. These ratios changed continuously during the first 12 hr after the administration when the polyclonal but not when the monoclonal RIA was used. In conclusion, blood concentrations assessed by the 3 methods are not identical, and when compared with the polyclonal RIA the monoclonal RIA exhibits 3 advantages: (1) much less crossreactivity with metabolites; (2) a constant RIA/ HPLC concentration ratio after the third hr after administration of CsA; and (3) a RIA/HPLC concentration ratio that is independent of the route of administration.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

In order to assess the effect of body weight on cyclo-sporine disposition, 45 adult uremic candidates for renal transplantation underwent detailed nutritional assessment and pharmacokinetic analysis. There were 10 obese and 35 nonobese patients defined as actual body weight (ABW) >125 Per cent of ideal body weight (IBW), and arm fat area > 90th percentile. There was no significant difference in demographic variables such as age, sex, number of diabetics, IBW, serum lipids, or liver function tests between the 2 groups. Although there was a significant difference in ABW, pharmacokinetic analyses failed to demonstrate significant differences in bioavailability, elimination half-life, clearance, or apparent steady state volume of distribution when these calculations were normalized by IBW, body surface area, or as absolute values. Multiple stepwise linear regression failed to demonstrate a significant correlation between serum lipids or body size measurements and these parameters. When dosed according to ABW, obese recipients of renal allografts had a mean serum RIA trough level of 227 ng/ml as compared to 121 ng/ ml in nonobese recipients on day 7. Therefore in order to achieve comparable drug concentrations in the early transplant period, CsA should be given to obese patients based on their IBW.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

Forty cyclosporine-treated renal allograft biopsies were stained with anti-Tac, a monoclonal antibody (MoAb) against interleukin 2 receptor (CD25), anti-transferrin receptor MoAb, and Ki67 MoAb (a proliferating cell marker), using a method of gold enhancement of the diaminobenzidine reaction product, in order to study the utility of these markers to differentiate rejection from other causes of graft dysfunction. Biopsies were selected on the basis of availability of sufficient frozen material and optical microscopy diagnosis, and only biopsies that showed acute cellular rejection and biopsies that did not show any sign of rejection (cyclosporine toxicity or a stable graft) were studied. In addition biopsies were stained with a panel of monoclonal antibodies reacting with CD45, CD3, CD4, CD8, EBM11 (a monocyte-macrophage marker), HLA-DR, DP, and DQ, using the usual indirect immunoperoxidase method. The clinical diagnosis (rejection versus no rejection) was made in ignorance of the biopsy findings.In 17 of 18 (94%) episodes of rejection, anti-Tac and anti—transferrin receptor—positive cells, and in 15 of 18 rejection episodes (83%), proliferating cells, were found in the interstitium. Anti-Tac—positive cells were never found in the 11 biopsies diagnosed as no rejection, and only in two a few anti—transferrin receptor—positive cells were present. The proportion of activated and proliferating cells in rejection episodes, expressed as the percentage of CD45-positive cells were: anti-Tac MoAb 4.1±2.9%, anti-transferrin receptor MoAb 6.9±3.9%, and Ki-67 MoAb 6.3±4.8%.There was a positive correlation between the total number of infiltrating cells and the number of cells stained with anti-Tac (r=0.75,P<0.005), anti-transferrin receptor MoAb (r=0.84,P<0.0001), and Ki-67 (r=0.50,P<0.05). The episodes of rejection that took place when cyclosporine levels were high had fewer 1L-2R-bearing cells in the interstitium than those that took place when cyclosporine levels were low (r=0.47,P<0.05).We conclude that during rejection the presence of activated and proliferating cells is a consistent finding, and that these markers could be useful in differentiating between rejection and other causes of graft dysfunction as well as in grading the severity of rejection.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

In human organ transplantation, the correct diagnosis of rejection may be difficult using clinical and/or histopathologic criteria, and immunologic assays should therefore be considered. We have applied monoclonal antibodies in 2-color flow cytometric (FACS) analysis to study phenotypic patterns of kidney-infiltrating activated and functional T and NK cell subsets in 132 post-transplantation biopsies. Viable intragraft lymphocytes and kidney tubular cells were obtained by the use of medium-sized ultra-sound guided needle biopsy. In uni-variate analyses, highly significant differences were observed between rejecting and stable grafts. During rejection, the proportions of HLA-DR+kidney tubular cells, total lymphocytes, T suppressor/cytotoxic cells, and natural killer (NK)-like cells increased; the fractions of phenotypically activated, HLA-DR+cells within the latter 2 cell populations were elevated. Further, during rejection phenotypic T suppressor inducer and T suppressor effector cells were virtually absent in kidney tissue but reappeared rapidly upon successful antirejec-tion therapy. Multivariate analyses revealed that combinations of antibodies in FACS analysis defining functional and activated subsubsets of T and NK cells allow a precise and rapid immunologic diagnosis of kidney rejection.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

In an analysis of over 4000 cyclosporine-treated recipients of second kidney transplants we observed a strong effect of HLA matching in living-related and cadaver transplants. In contrast to the results obtained in first cadaver transplants, second cadaver transplants benefited substantially from matching for HLA-A locus antigens. The strongest effect of matching was found when HLA-A, HLA-B, and HLA-DR antigens were analyzed together: 214 second grafts with no mismatch had a survival rate of 82 ± 3% at two years in contrast to a 49 ± 4% rate in 149 grafts with 6 mismatches (P<0.0001, log rank). Patients whose first graft functioned for more than 1 year had a significantly higher second graft survival rate than patients with shorter first graft duration. Because the effect of HLA matching is particularly strong in patients with <1 year first graft duration, it is suggested that HLA well-matched kidneys should be allocated to them with priority.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

Blood samples were obtained, at the time of organ donation, from 31 consecutive brain-stem-dead (BSD) donors referred to one transplant coordinator during a 9-month period. Twenty-four cases (77%) had clinical diabetes insipidus (DI), which was poorly controlled with marked dehydration in a majority of cases (serum osmolality range 268–357; median 302 mOSM/kg). Serum triiodothyronine (T3) was subnormal in 25 (81%); all had normal or high serum reverse T3; and the serum free thyroxine (T4) index was subnormal in 9 (29%), and TSH was subnormal in 7 (23%). In no case were T4and TSH both subnormal and results were typical of the sick euthyroid syndrome rather than TSH deficiency. Of 21 cases not receiving corticosteroids, 5 (24%) had a serum cortisol above 550 nmol/L (20 μg/dl), excluding ACTH deficiency, and only 1 had undetectable cortisol levels. Those with severe hypotension did not have significantly lower serum cortisol (mean 354 vs. 416;P>0.5). Levels of prolactin, growth hormone, gonadotrophins, and gonadal steroids were variable, but only a minority were frankly deficient in these hormones. BSD donors frequently have DI, which is often managed poorly by nonspecialists and requires appropriate replacement therapy. In contrast most patients are not totally deficient in anterior pituitary hormones. Routine hormonal therapy with cortisol and T3cannot, therefore, be justified on endocrinological grounds. Widespread introduction of such treatment should only follow controlled trials that clearly demonstrate clinically significant improvement in the transplanted organ function, without detriment to the donor.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

HIV-1 antibody determination by ELISA screening takes 4 hr to complete and is not reliable. Regular Western blot can take up to 24 hr. This makes organ transplantation both difficult and risky with regard to HIV-1 transmission. We developed a modification of the Western blot technique that takes under 50 min, is transportable, is definitive on site, and does not delay organ retrieval. This test has been called the Quick Western Blot. We examined 459 serum specimens from referrals; from the AIDS Testing Proficiency Panel, Walter Reed Army Institute of Research; and from 36 organ donors. All specimens were tested by ELISA HIV-1 Ab screening, the regular Western blot, and by the Quick Western Blot. The organ donors were initially tested on-site during organ procurement by the Quick Western Blot and later had complete testing by the reference methods. Compared with regular Western blot, the ELISA showed a specificity of 78.4% and a positive predictive value of 65.5%, whereas the Quick Western Blot was as reliable and specific as the regular Western blot, but much quicker. Because of the rapidity and specificity of the test, this test has particular utility in the screening of organ donors, as was shown in a case of multiple organ donation where the ELISA was negative, but the Quick Western Blot was found positive and thereby prevented the donation of HIV-1 infected organs.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

T cell clones (n=456) were derived from 9 patients following allogeneic bone marrow transplantation (BMT) with or without acute graft versus host disease (aGVHD) and from 4 healthy donors. The cloning efficiency was 63.2% in controls, 13.2% and 12.1% in patients with or without aGVHD. Once established, T cell clones were typed for surface markers (CD3, CD4, CD8) and tested for production of IL-2 and expression of cytolytic activities in a lectin-dependent cellular cyto-toxicity assay (LDCC) and against the K562 target cell line to detect natural killer activity. We found the expected imbalance of CD4/CD8 clones in BMT patients, as compared to controls. A higher proportion of IL-2-producing clones was observed in patients with aGVHD (83.5%;P<0.02) as compared to patients without aGVHD (64.8%) and controls (68.5%). No major differences were found in terms of LDCC, whereas an increased percentage of clones with NK-like activity was found in patients with aGVHD (34.7%,P<0.05) as compared to patients without aGVHD (29.5%) and controls (21.3%).The clones were also tested for inhibition of IL-2 production mediated by cyclosporine. Such inhibition could be obtained in virtually all clones both in patients with or without aGVHD, suggesting that the latter is probably not due to the emergence of CsA-resistant clones.In conclusion, this study demonstrates a low cloning efficiency in BMT patients associated with the well-known CD4/CD8 imbalance. A higher production of IL-2, an increased NK activity, but not the presence of CsA-resistant clones appear to differentiate patients with from patients without aGVHD.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

To study the mechanism of induction of specific un-responsiveness to allografts in animals treated with antilymphocyte serum and donor bone marrow cells, we examined the effect of donor BMC on second-set graft rejection responses caused by antidonor-sensitized spleen cells (SSC) in mice bearing donor ear skin grafts. Anti-C3H SSC were obtained from skin-grafted, ALS-treated B6AF1mice after rejection of their grafts. The second-set rejections of C3H skin grafts were assessed in B6AF1mice following adoptive i.v. transfer of 30×106SSC one day after grafting. Median survival time (MST) of C3H skin grafts in the group injected with SSC was 7 days, which is significantly lower than an MST of 11 days observed in the control group, which exhibited a first-set graft rejection response. Addition of 50×106C3H BMC to 30×106SSC abrogated the second-set rejection of C3H skin grafts (MST =11 days). This abrogation effect of BMC is strain-specific, since BMC of a third-party strain (DBA/2) failed to abrogate the second-set rejection responses caused by anti-C3H SSC. Of the different C3H lymphoid cells tested for abrogation of the second-set graft rejection, BMC were the most effective. Splenocytes were more effective than thymocytes, which showed a partial effect. Lymph node cells had no effect. Our data suggest that unresponsiveness to allografts in animals treated with the ALS/bone marrow protocol may result from the inactivation of graft-reactive cells by donor BMC.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID

摘要:

In this study we examined the effect of cyclosporine on three distinct subsets of T suppressor (Ts) cells identified in a rat renal allograft model. Ts inducer (Ts1) cells having the CD4 marker are found in the spleens of DA rats undergoing acute rejection of LEW kidneys. Transducer (Ts2) and effector (Ts3) cells both carry the CD8 marker and are found in the spleens of long-term surviving DA rats bearing LEW kidney allografts made tolerant by donor-specific blood transfusions or by cyclosporine (in most cases). These latter cells are distinguished by their susceptibility to cyclophosphamide (CY), Ts2 cells being resistant while Ts3 cells are sensitive to CY. When Ts cells from DA rats undergoing acute graft rejection of LEW kidneys or bearing long-term-surviving LEW kidneys that had been treated with cyclosporine (10mg/kg/day) for 2 or 10 days, respectively, were adoptively transferred into lightly irradiated DA recipients, these cells were still able to specifically induce long-term survival of LEW kidneys.

ISSN:0041-1337    

出版商:OVID    

年代:1989

数据来源: OVID