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S V Fuggle

Publications and source records attributed to S V Fuggle.

At least 37 records · Page 2Linked to original sources
PubMed1998

Apoptosis in ischemia/reperfusion injury of human renal allografts.

BACKGROUND: Ischemia/reperfusion injury of human renal allografts has a number of clinically significant consequences. A number of mechanisms of ischemia/ reperfusion injury have been elucidated, and there is evidence that apoptosis may be a contributing factor. METHODS: To examine immediate posttransplant events, fixed tissue sections from paraffin-embedded wedge biopsy specimens taken before and after reperfusion of human renal allografts were stained using terminal deoxytransferase-mediated dUTP nick-end labeling to detect the DNA fragmentation characteristic of apoptosis. Thirty-six pairs of pre- and postreperfusion biopsy specimens were examined, 11 from living-related donor renal transplants and 25 from cadaveric donor transplants. RESULTS: Quantitation of the terminal deoxytransferase-mediated dUTP nick-end labeling signal showed that significantly more apoptosis occurred in postreperfusion compared with prereperfusion biopsy specimens from cadaveric donor transplants, but a similar difference was not observed in living-related donor renal transplants. Furthermore, significantly more apoptosis was observed in postreperfusion biopsy specimens from cadaveric compared with living-related renal transplants. Postreperfusion biopsy specimens from kidneys that were cold preserved longer than 30 hr had a higher mean apoptosis score than those stored for less than 24 hr, but the result was not statistically significant. CONCLUSIONS: Thus, apoptosis occurs predominantly as a result of reperfusion after cold preservation of cadaveric donor renal allografts and provides additional information regarding the extent of ischemia/ reperfusion injury in an organ. The clinical value of this information remains to be determined.

Adolescent↗
PubMed1998

Detection of microchimerism after allogeneic blood transfusion using nested polymerase chain reaction amplification with sequence-specific primers (PCR-SSP): a cautionary tale.

In bone marrow transplantation, the detection of chimerism is an important adjunct to the repertoire of tests available for determining acceptance of the graft. In solid organ transplantation, there is currently intense interest in the role that chimerism plays in both short- and long-term host reactivity to the graft. Allogeneic blood transfusion has been associated with a subtle immunosuppressive effect in renal transplantation and chimerism is implicated as a possible mechanism for this effect. To assess the survival of allogeneic cells after blood transfusion or transplantation, we have developed a technique based on molecular typing for HLA class II alleles, which enables the detection of donor-derived cells in patients receiving blood transfusions. While developing this technology, we investigated why we and others observe false amplification. Sequencing of false products has shown that they arise from amplification of both pseudogenes and non-pseudogenes present in the DNA under test. Elucidation of this phenomenon allows the amplification of these false products to be predicted in any given combination and hence avoided by the judicious selection of primers. Validation has been achieved by following donor alleles after transfusion of blood containing defined numbers of leukocytes expressing selected mismatched antigens.

Blood Donors↗
PubMed1998

Cell adhesion molecules in clinical renal transplantation.

Leukocyte adhesion molecules are critically involved at a number of stages in immune and inflammatory responses, and their importance in the response to a renal allograft has been recognized for some years. They are involved in antigen presentation, in the cascade of events leading to extravasation of leukocytes into the allograft, in the subsequent migration of leukocytes through the extracellular matrix, and in the interactions between effector and target cells. Thus the adhesion molecules are highly attractive targets for therapeutic intervention in organ transplantation. Strategies have been explored to exploit the involvement of adhesion molecules in ischemia/reperfusion injury, allograft rejection, and the induction of immunological tolerance. Furthermore, the expression of a number of adhesion molecules is regulated by cytokines, and elevated levels may be detected both in transplant biopsies and as soluble forms measured in serum and urine. It has been proposed that these changes in levels might provide useful information in the diagnosis of allograft rejection and differentiation from other causes of graft dysfunction.

Animals↗
PubMed1998

Ischemia/reperfusion injury in human kidney transplantation: an immunohistochemical analysis of changes after reperfusion.

Organs used for transplantation undergo varying degrees of cold ischemia and reperfusion injury after transplantation. In renal transplantation, prolonged cold ischemia is strongly associated with delayed graft function, an event that contributes to inferior graft survival. At present, the pathophysiological changes associated with ischemia/reperfusion injury in clinical renal transplantation are poorly understood. We have performed an immunohistochemical analysis of pre- and postreperfusion biopsies obtained from cadaver (n = 55) and living/related donor (LRD) (n = 11) renal allografts using antibodies to adhesion molecules and leukocyte markers to investigate the intragraft changes after cold preservation and reperfusion. Neutrophil infiltration and P-selectin expression were detected after reperfusion in 29 of 55 (53%) and 24 of 55 (44%) cadaver renal allografts, respectively. In marked contrast, neutrophil infiltration was not observed in LRD allografts, and only 1 of 11 (9%) had an increased level of P-selectin after reperfusion. Immunofluorescent double-staining demonstrated that P-selectin expression resulted from platelet deposition and not from endothelial activation. No statistically significant association was observed between neutrophil infiltration and P-selectin expression in the glomeruli or intertubular capillaries despite the large number of cadaver renal allografts with postreperfusion changes. Neutrophil infiltration into the glomeruli was significantly associated with long cold ischemia times and delayed graft function. Elevated serum creatinine levels at 3 and 6 months after transplantation were also associated with the presence of neutrophils and platelets after reperfusion. Our results suggest that graft function may be influenced by early inflammatory events after reperfusion, which can be targeted for future therapeutic intervention.

Adult↗
PubMed1998

Identification of an HLA-C variant allele, Cw*0805, by sequencing based typing.

We have identified a variant HLA-C allele, Cw*0805, in DNA isolated from a blood sample used in the UK National External Quality Assessment Scheme for Histocompatibility and Immunogenetics. The DNA was initially typed by PCR-SSP and PCR-SBT as Cw*0802, 1602 with a single mismatch in exon 2 of one of the alleles. The two HLA-C alleles were separated by locus- and allele-specific PCRs and sequencing analysis showed that the Cw*0802 allele was variant in this sample, with a single mismatch in exon 2 at position 289.

Alleles↗
PubMed1998

A new national allocation scheme for adult kidneys in the United Kingdom. United Kingdom Transplant Support Service Authority (UKTSSA) Users' Kidney Advisory Group and its Task Forces.

A new allocation scheme for kidneys from adult cadaver donors was introduced in the UK on July 1st, 1998. The new scheme is based on data from an analysis of factors influencing transplant survival instigated by the Kidney Advisory Group (KAG) of the UKTSSA. A cohort of 6,363 first cadaver allografts performed in the UK between 1986-1993 was used for the analysis with 99.6% one-year follow-up and 97.8% 5-year follow-up. HLA matching was one of a number of factors that were found to influence transplant survival, thus supporting the policy of exchange of kidneys based on matching. The new allocation scheme is based on 3 tiers in which cadaver organs are offered first in Tier 1 to patients with zero HLA antigen mismatches (000 matchgrade), in Tier 2 to favorably matched patients, (matched for HLA-DR and mismatched for a maximum of one HLA-A and one-B locus antigen -100, 010, 110 matchgrades) and remaining kidneys in Tier 3 to non-favorably matched patients. A points score devised by a subgroup of the KAG to reflect natural justice and common sense is used as a discriminator between equally matched patients. The points are based on recipient age, donor-recipient age difference, waiting time, matchability for HLA antigens, sensitization to HLA antigens and the transplant unit balance of organ exchange. The performance of the scheme will be closely monitored, but computer simulations predict that there will be an overall improvement in transplant survival as a result of an increase in well matched transplants.

Adolescent↗
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↗
PubMed1996

Late reflush in clinical renal transplantation. Protection against delayed graft function not observed.

Mechanical flushing of cadaveric kidneys with organ preservation fluid immediately before transplantation has been reported to be associated with improved early graft function. We report here the results of a prospective randomized controlled study of cadaveric renal transplantation after late reflush with organ preservation fluid in which no benefit with respect to delayed graft function was observed and, indeed, the protocol may have been harmful. The study was terminated after recruitment of only 18 patients (9 to each arm) because postreperfusion biopsies of reflushed kidneys contained unusual features, including abnormal cellular debris within the tubules or eosinophilic proteinaceous material within Bowman's capsule. These features were not present in the control kidneys. Acute tubular necrosis and biopsy-proven acute rejection episodes were more frequently seen in the reflushed kidneys, but at 1 year there was no significant difference in the function of the surviving grafts.

Adult↗
PubMed1996

Expression of granzyme A and perforin in mouse heart transplants immunosuppressed with donor-specific transfusion and anti-CD4 monoclonal antibody.

Granzyme A and perforin are produced by activated cytotoxic T lymphocytes and their expression correlates with the appearance of cytotoxicity. Using in situ hybridization and immunohistochemistry, we examined the phenotype of cellular infiltration and the appearance of granzyme A+ and perforin+ cells in a mouse cardiac transplant model where the recipients were pretreated with donor-specific transfusion, anti-CD4 monoclonal antibody, or both. While the profiles of cellular infiltration failed to correlate with graft survival, tolerized grafts, as compared with untreated allografts, showed a decreased frequency of granzyme A and perforin expression. These functional markers of cytotoxic T lymphocytes can differentiate between rejecting and indefinitely surviving grafts and may be of value in dissecting the immunological events involved in tolerance induction.

Acute Disease↗
PubMed1993

Variation in expression of endothelial adhesion molecules in pretransplant and transplanted kidneys--correlation with intragraft events.

Endothelial adhesion molecules are directly involved in the localization and migration of leukocytes from the circulation into tissues at sites of inflammation. We have compared the expression of PECAM-1 (CD31), ELAM-1, ICAM-1 (CD54), and VCAM-1 in pretransplant (n = 20) and needle-core biopsies from renal transplants obtained during different clinical circumstances (n = 42). PECAM-1 was consistently expressed on all endothelium in both pretransplant and transplant biopsies. In contrast, there was variation in endothelial expression of ELAM-1 and in proximal tubular expression of ICAM-1 and VCAM-1 between pretransplant biopsies. After transplantation induced expression of endothelial ELAM-1 and VCAM-1 and tubular induction of ICAM-1 and VCAM-1 was detected. Induced adhesion molecule expression was frequently associated with focal leukocyte infiltration, and there was a significantly higher level of CD45 and CD25 positive cell infiltration in biopsies with induced adhesion molecule expression. The induction of adhesion molecule expression is evidence of endothelial activation in these transplant biopsies. Comparison of adhesion molecule expression and HLA-class II antigen expression revealed that induced tubular class II antigens may be detected in the absence of induced adhesion molecule expression.

Antigens, Differentiation, Myelomonocytic↗
PubMed1993

In situ cDNA polymerase chain reaction. A novel technique for detecting mRNA expression.

We report a novel method for detecting intracellular messenger RNA by combining the techniques of in situ hybridization and polymerase chain reaction (PCR) (in situ cDNA PCR). The technique could detect low abundancy signals and distinguish different levels of gene expression. We examined the expression of the functional markers of activated cytotoxic T lymphocytes, granzyme A, and perforin in human lymphocytes from in vitro cultures. The amplification products were found in the cells and the supernatants, with the distribution critically affected by the protease digestion conditions. The specificity of amplification was confirmed by electrophoretic analysis and Southern blotting. We conclude that the in situ cDNA PCR technique offers a sensitive method of measuring the frequency of signal-expressing cells and has significant research and clinical applications.

Biopsy↗
PubMed1991

Immunophenotypic analysis of leukocyte infiltration in the renal transplant.

Immunohistological analyses of transplant biopsy material can provide valuable insight into the intragraft events following transplantation. The magnitude, composition and phenotype of the leukocyte infiltration and up-regulation of parenchymal antigen expression within the renal allograft have been major areas of interest. Immunohistological analyses have demonstrated the presence of leukocyte infiltration not only during clinical rejection episodes, but also during periods of stable graft function. Nevertheless the infiltration is significantly increased during clinical rejection. Indeed, quantitation of the leukocyte infiltration using a morphometric point counting technique or image analysis can be used prospectively to diagnose rejection in the renal transplant recipient. Macrophages and T lymphocytes (both of the CD4 and CD8 phenotype) are the major infiltrating cells, B lymphocytes and NK cells being present, but in the minority. Activated and proliferating cells have been demonstrated during allograft rejection and may provide a means of identifying a functionally active infiltrate.

Antibodies, Monoclonal↗
PubMed1989

Monoclonal antibody definition of the DRB3 allele, HLA-Dw25.

Monoclonal antibodies are powerful tools for analyzing HLA antigen polymorphism. We have investigated the serological and biochemical nature of the DRw52-related antigen defined by the monoclonal antibody NDS10. A detailed analysis of the population distribution of NDS10 reactivity revealed that the epitope was present on a subpopulation of DRw52 positive cells. A distinct pattern of reactivity was found within DR3 individuals: all of the B18,DR3 cells were NDS10 positive, whereas the A1,B8,DR3 cells were negative. All of the DR5(w11) cells and two of three DRw12 cells reacted with NDS10. NDS10 reactivity with DRw6 was not restricted to either of the serologically defined subtypes; three of 17 DRw13 and nine of 10 DRw14 cells were NDS10 positive. NDS10 was unreactive with all of the DRw8 cells tested. Two-dimensional gel analyses revealed that the NDS10 molecule precipitated from DR3, DR5(w11) and DRw6(w14) cell lines had an identical beta chain profile. These data indicate that NDS10 recognises the Dw25 allele of the DRw52 complex.

Antibodies, Monoclonal↗
PubMed1989

Immune responses to the circumsporozoite protein of Plasmodium falciparum in relation to HLA-DR type.

Associations between HLA-DR/DQ phenotypes and immune responses to the circumsporozoite (CS) protein of the human malaria parasite, Plasmodium falciparum, were investigated in Thai adults. The frequency of DR and DQ types did not differ between 15 high antibody responders and 16 antibody non-responders. Initial results which suggested that lymphocyte responses to a CS-derived peptide (R32tet32) might be HLA-DR associated were not confirmed by subsequent extension of the study. Thus, MHC restriction appears not to be a major factor in the frequently observed unresponsiveness of the immune system to the P. falciparum sporozoite. Based on these results, it is expected that MHC restriction will not be a major obstacle to the application of a malaria vaccine in outbred human populations.

Adult↗
PubMed1989

Biochemically detected HLA-DQ polymorphism in DR-matched donors and recipients of renal allografts.

We performed a retrospective biochemical analysis of HLA-D-region antigens of serologically DR-compatible donors and recipients of renal allografts. No incompatible D-region antigens were detected in grafts with a stable clinical course--i.e., there were no rejection episodes--whereas incompatibility for one or more D-region antigens was found in all 13 grafts with rejection. Thus, mismatched D-region antigens may provide a stimulus for early rejection in these grafts.

Cell Line↗