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Results for “Histocompatibility Testing”

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At least 379 records · Page 21Linked to original sources
PubMed1986

Prediction of crossmatch outcome in highly sensitized dialysis patients based on the identification of serum HLA antibodies.

High levels of allosensitization (greater than 50%), which often occur in dialysis patients awaiting renal transplant, make donor selection difficult. Such patients may be included in elaborate protocols in which they are crossmatched with all available ABO compatible donors, or crossmatching may be deferred until a very-well-matched donor becomes available. The former approach of random crossmatching is costly and inefficient, while the latter approach may overlook crossmatch-compatible donors. We believe that the identification of antibodies present in highly reactive sera and the use of this information in donor selection would increase the frequency of crossmatch-negative donors for these patients. In this study eleven sera, reactive with 70% to 100% of a random cell panel, were obtained from multiply transfused dialysis patients. Sera were analyzed by standard (CDC) and antiglobulin augmented (AHG-CDC) lymphocytotoxicity, and by differential absorption with HLA-typed platelets. All sera contained only one or two antibodies directed against the high frequency public HLA epitopes, accounting for 85% to 100% of each serum's total reactivity. These characterized sera were crossmatched with 114 random normal donors. The frequency of negative crossmatches was 20.5%. However, if the serum antibody data had been used to preselect donors for crossmatch--that is, to exclude donors that were likely to be positive--the negative crossmatch frequency would have increased to 86.4%. The use of the serum analysis data in donor selection would have reduced the total number of required crossmatches by 78%. Serum analysis correctly predicted the outcome of 95.6% of crossmatches performed with an average of 3% false positives and 1.3% false negatives. This approach to donor selection reduces unnecessary crossmatching and increases the likelihood of finding crossmatch-compatible donors for highly reactive patients.

ABO Blood-Group System↗
PubMed1990

The incidence of antimonocyte alloantibodies. Evidence that prospective crossmatching is unnecessary.

Monocyte cytotoxic crossmatches were performed on the sera of all 164 patients on the Victorian transplant waiting list in order to determine the incidence of antimonocyte antibodies and to assess if prospective screening for these antibodies would be justified. Initially all current sera were screened without absorption by the cytotoxicity assay for the presence of antilymphocyte and antimonocyte reactivity against a panel of ten donors. Subsequently, a further 45 "peak"-reactive sera were screened against the same panel. Forty sera with monocyte specific reactivity were selected. MHC class I and class II antibodies were absorbed with pooled platelets and B lymphoblastoid lines, respectively. The absorbed sera were then screened against T cells, B cells, and monocytes of 38 donors for monocyte-specific antibodies. Seven sera from six recipients showed monocyte reactivity without T cell or B cell reactivity, with panel reactivity ranging from 3-29%. This gives an overall patient incidence of 3.6%, and when expressed as a percentage of consecutive crossmatches has a calculated incidence of 0.45%. This incidence is much lower than previously reported and would indicate that prospective crossmatching for antimonocyte antibodies would not be of benefit in this group of patients.

B-Lymphocytes↗
PubMed1997

HLA antibody screening: comparison of a solid phase enzyme-linked immunoassay with antiglobulin-augmented lymphocytotoxicity.

BACKGROUND: IgG antibodies to HLA class I antigens can cause hyperacute rejection of renal allografts. Screening of sera from such transplant candidates is laborious, time-consuming, and expensive when performed by sensitive antihuman globulin-augmented lymphocytotoxicity (AHG-CDC). METHODS: Because 60-70% of our transplant screens are negative, we evaluated a solid phase enzyme-linked method (EIA) as a potential prescreen by parallel testing 215 sera by AHG-CDC and by EIA. This EIA method is designed to detect only IgG antibodies, and all positive AHG-CDC sera were retested after dithiothreitol treatment. RESULTS: There was 96.2% concordance between the tests for IgG antibodies. Seven sera (3.25%) were positive by EIA alone, and one (0.46%) was negative by EIA alone. The EIA method was also less costly ($15.00 versus $105.00) and less time consuming (hours versus days) than AHG-CDC panel testing for large numbers of sera. CONCLUSIONS: We conclude that this EIA method is simple, sensitive, objective, and cost effective as a prescreen for HLA class I antibodies.

Cytotoxicity Tests, Immunologic↗
PubMed2000

The relationship between acute rejection and chronic rejection is highly dependent on specific MHC matching: a multi-strain rat heterotopic heart transplant study.

BACKGROUND: The impact of acute rejection, immunosuppression, and infection, specifically cytomegalovirus infection, on the development of chronic rejection in the cardiac allograft, has been the subject of a large number of investigations. One of the difficulties in finding associations has been the marked immunologic heterogeneity of the patient population coupled with the lack of the ability to HLA match. Furthermore, the ideal animal model, which duplicates as well as controls for this immunologic heterogeneity, is lacking. METHODS: To try to simulate differences in HLA matching, immunosuppression regiments and cytomegalovirus infection, heterotopic heart transplantation was performed in two separate, complete MHC mismatch, rat strain combinations (WF-LEW, BN-LEW) requiring chronic immunosuppression and employing four separate immunosuppression/infection protocols. Animals were followed for 6 months, killed, and rejection and vascular changes were scored blinded to the group. RESULTS: The mean vascular and acute rejection scores were not significantly different between treatment regiments for either specific strain combination. There was a trend for the subtherapeutic groups to have higher vascular scores. Overall, there were no significant differences in vascular scores between the WF-LEW and BN-LEW groups (1.25+/-0.18 vs. 1.13+/-0.20, P=NS). Similar numbers of WF-LEW and BN-LEW exhibited cellular infiltration and necrosis of the allograft, but the intensity of the response (rejection score) was more severe in the WF-LEW combination (4.54+/-0.22 vs. 3.92+/-0.21, P=0.052) when limiting the analysis to those with myocyte necrosis. There was no significant correlation between acute rejection and vascular lesion severity in the WF-LEW combination (r=0.22, P=NS) but a high correlation between these parameters in the BN-LEW combination (r=0.74, P<0.0001). CONCLUSIONS: These data suggest that, although acute rejection and chronic rejection are related, MHC matching may influence their interdependence. These data also may explain why the clinical association between acute and chronic rejection is difficult to demonstrate.

Acute Disease↗
PubMed1988

Characterization of class I bovine lymphocyte antigens (BoLA) by one-dimensional isoelectrofocusing.

BoLA class I antigens were characterized in a group of British and Dutch Friesian cattle by one-dimensional isoelectric focusing (1D-IEF) and the results compared with serology using alloantisera and microcytotoxicity. For IEF analysis, non-stimulated peripheral blood mononuclear cells (PBM) were metabolically labelled with 35S methionine, detergent lysates were prepared and MHC molecules precipitated with the monoclonal antibodies (mAbs) W6/32 or B1.1G6. Staphylococcus protein A precipitated antigens were separated on a vertical slab gel under denaturing conditions. The banding patterns seen for the W6/32 precipitated molecules obtained by 1D-IEF were compared with the serological specificities. Characteristic banding patterns were observed for most serological specificities as well as workshop undefined haplotypes. These patterns were seen both in families and the outbred population. In families IEF haplotypes segregated with serotypes. Additional MHC class I products were suggested by variable banding patterns for different w10 haplotypes and when using the different mAbs. A pulse chase experiment with a w12 animal also suggested more than one expressed product. The w2 and w5 specificities were not precipitated by either W6/32 or B1.1G6 and w6.2 and w6.4 were precipitated by W6/32 but not by B1.1G6. These results show that 1D-IEF is useful for BoLA typing. For the characterization of class I antigens, however, much depends on the mAbs used.

Animals↗
PubMed1991

Expression of MHC class I and II molecules by cadaver retinal pigment epithelium cells: optimization of post-mortem HLA typing.

The objective of this study was to investigate the expression of MHC antigens by retinal pigment epithelium cells (RPE) after stimulation with interferon-gamma (IFN-gamma) and to improve the currently practised technique of cadaver HLA typing. A concentration of 100 U/ml IFN-gamma induced expression of class I molecules up to greater than 90% 3 days after stimulation, whereas 50 U/ml were required for the expression of HLA-DR to greater than 90%. A concentration of 750 U/ml induced 35-45% expression of HLA-DP and less than 25% HLA-DQ after 3 days. Cells were serologically typed using the standard lymphocytotoxicity assay 3 days after stimulation with 250 U/ml IFN-gamma. Typing of class I specificities was complemented by one-dimensional isoelectric focusing (1D-IEF). We observed high concordance between the results of the RPE typing and the lymphocytotoxicity test on the same donors. Our results show complete typing of class I and II antigens post-mortem, which, in particular, enables graft matching and improvement of graft survival in recipients of organs removed many hours after death such as the cornea.

Adolescent↗
PubMed1975

A new alloantigenic system on human lymphocytes.

Certain otherwise well-definedHL-Atyping sera contain extra antibodies which do not appear to be closely related to known HL-A specificites and which react in the cytotoxic test with chronic lymphaticleukemia lymphocytes but not with normal lymphocytes. Despite nonreactivity in thecytotoxic test, absorption studies show that the antigenic factors are well represented onnormal lymphocytes. Based on cross-absorption experiments using both normal andleukemic cells, followed by back-testing with the cytotoxic test against a leukemiccell panel, evidence is presented suggesting the existence of a complex, probablymultiple allelic system on human lymphoid cells which appears distinct from the, HL-Asystem, and is definable by serologic techniques.

Cross Reactions↗