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Results for “Histocompatibility Testing”

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At least 163 records · Page 9Linked to original sources
PubMed1971

Marrow grafts between canine siblings matched by serotyping and mixed leukocyte culture.

Marrow grafts were carried out between 16 canine sibling donor-recipient pairs. The pairs were matched by serological histocompatibility testing and were nonreactive in a one-way mixed leukocyte culture. Recipients were prepared for transplantation by 1500-1580 R of total body irradiation. Donor marrow was infused within 4 hr of irradiation. Recipients were not given immunosuppressive drug therapy after grafting. All 16 recipients showed evidence of prompt and sustained allogeneic marrow engraftment. Six died between 30 and 128 days after grafting with graft-versus-host disease, and three died between days 72 and 230 with pneumonia but no evidence of graft-versus-host disease with the exception of lymphoid atrophy. Seven recipients survived without graft-versus-host disease and are in excellent health between 200 and 684 after grafting. In summary, fatal graft-versus-host disease was observed in a number of canine recipients despite matching with their sibling donor by serological histocompatibility testing and by mixed leukocyte culture in a manner similar to that employed to define human HL-A matched sibling pairs. The graft-versus-host disease in these matched siblings developed more slowly than that observed in mismatched dogs, but the ultimate death of approximately half of the matched recipients emphasizes the need for posttransplantation immunosuppression even in this "compatible" situation.

Animals↗
PubMed1979

A rapid method for preimmunisation and grafting of mice with ear skin.

A schedule for a histocompatibility test among mice involving preimmunisation with ear skin is described and has been tested with the weak transplantation antigen H-Y in C57BL responder and C3H nonresponder strains. The transplantation technique was tested by autografting, and more than 99% of the operations were successful. The priming method reduces the survival of ear skin grafts in every combination tested.

Animals↗
PubMed2000

[Comparison of the results of HLA typing using serologic and molecular genetics methods].

A comparison of the HLA class I typing in 50 unrelated individuals by means of serological and molecular genetic (PCR-SSP) methods was carried out. DNA-typing is more fast and reliable method in comparison with serology. It is necessary to introduce molecular genetic methods for the detection of HLA class I alleles. On the other hand there are alleles, which are not expressed on cell surface. In our laboratory both methods are established and the results of both were compared. It may be useful for determining the selection strategy of HLA-identical donor-recipient pair suitable for bone marrow transplantation. The results demonstrated 9% misassignments of HLA-A antigens by serology, 11% of HLA-B and 39% of HLA-C. The serological discrepancies found were of three categories: false negatives, false positives, and an incomplete typing. The vast majority of the discrepancies were due to a combination of relatively low expression of HLA antigens, lack of serological reagents and misclassification of antigens within cross-reactive groups. These results indicate that nowadays the serological typing is insufficient for clinical histocompatibility testing. (Tab. 3, Ref. 16.)

Alleles↗
PubMed1992

Results of allogeneic bone marrow transplantation with unrelated or mismatched donors.

As most patients are not fortunate enough to have an HLA-matched sibling to use as a bone marrow donor, attention has focused on the use of either HLA-matched but unrelated donors or HLA-mismatched family members. With the maturation of the field of histocompatibility testing, it is now possible to quantitate with relative precision the degree of disparity between patient and donor. In general, it appears that with respect to histocompatibility differences between donors other than HLA-matched siblings, there is an increased incidence of acute graft-versus-host disease, with the risk correlated with the degree of histoincompatibility. However, the overall disease-free survival is not always adversely affected, as a graft-versus-leukemia effect may counterbalance the increased death rate from graft-versus-host disease. To find donors for most patients, efforts are under way to recruit a large number of unrelated volunteers into the National Marrow Donor Program.

Bone Marrow Transplantation↗
PubMed1975

Two techniques to make the lymphocytotoxic crossmatch more sensitive: prolonged incubation and the antiglobulin test.

Two techniques capable of increasing the sensitivity of the lymphocytotoxic crossmatch test were compared for sensitivity and practically using the fluorochromasia modification of the cytotoxicity test. Prolonging the incubation period after addition of complement in the standard test resulted in further positive reactions. Modification of the test by addition of antiglobulin reagent increased the reactivity more than prolonged incubation of the standard test. The fluorochromasia adaptation of the antiglobulin test had sensitivity equivalent to the original antiglobulin method. The antiglobulin technique had a number of disadvantages, and neither the prolonged incubation nor the antiglobulin procedure meet the requirements of a busy histocompatibility testing service in view of the time consumed for the first and the complexity of the second.

Coombs Test↗
PubMed1975

[SLA-testing and MCL-reaction in swine and their reaction to rejection of skin and liver transplants].

After porcine skin allografts there is a regular rejection within 6 - 12 days with a characteristic pattern of humoral antibodies corresponding to the histocompatibility testing. The rejection of skin allografts may be due to weak histocompatibility antigens which are not measured by the SL-A and MLC typing. A porcine liver allograft induces an immunological reaction of moderate degree. This state of hyporeactivity i. e. partial tolerance could be induced and maintained by the release of tolerogenic antigens from the porcine liver.

Animals↗
PubMed2000

The relevance of proficiency testing for laboratories involved in cadaveric organ transplantation and its consequences for graft survival.

Organ exchange organizations such as Eurotransplant allocate organs on the basis of histocompatibility testing results. For this reason it is essential that all data reported by the affiliated laboratories are accurate and reliable. The Eurotransplant Reference Laboratory (ETRL) organizes proficiency testing schemes for the tissue-typing centers of the respective renal transplantation units participating in Eurotransplant. Each year, the ETRL sends out 8 peripheral blood samples of healthy blood donors for serological typing and crossmatching, 16 sera to screen for the presence and definition of HLA alloantibodies and 20 DNA samples for molecular typing to the 49 participating centers. The results are collected centrally and reported back to the participants in an open way. These exercises show that the quality of HLA typing, screening and crossmatching improved significantly over the years. In particular, the introduction of molecular typing for HLA-DR resulted in an increase of reliability. The clinical relevance of a reliable HLA typing was demonstrated in a selected group of transplants, the zero HLA-A,-B,-DR- mismatched group. After retyping the donors, 146 of the 3,458 matched transplants appeared to have a mismatch and those transplants had a significantly lower graft survival rate. A continuing problem, however, is the result of screening for panel reactive antibodies (PRA), where the percentage PRA reported for each serum varies significantly from center to center. The results indicate that the use of a PRA value for classification of patients and allocation of organs should be revisited.

Cadaver↗
PubMed1995

Evaluation of HLA antibodies with the PRA-STAT test. An ELISA test using soluble HLA class I molecules.

HLA-specific antibody, present before or after transplantation, may adversely effect graft outcome. Antibody testing by cytotoxicity (CYT) is laborious, requires viable lymphocytes, does not differentiate non-HLA cytotoxic antibody, and cannot be used readily on specimens from patients being treated with cytotoxic antibodies. We have evaluated PRA-STAT, an antibody screening kit that uses an ELISA test with soluble HLA class I molecules as targets. We performed 219 tests on a variety of serum specimens, 128 of which were also tested by CYT. There was a highly significant correlation (r = 0.78, P < 0.001) between PRA-STAT (PS) and CYT for the detection of IgG antibodies. Of 66 sera reactive in both assays, 18% had identical specificities defined in both, 27% were more reactive in PS than in CYT, 8% were more reactive in CYT, and 47% had different specificities in the 2 assays, with overlap in slightly more than half the cases. Of 13 sera reactive only in PS, 2 were from non-transfused, nontransplanted males with no evidence of lymphocyte-reactive antibody by antiglobulin tests. PS uses an IgG-specific conjugate, therefore IgM class I-specific antibodies cannot be identified--however, their presence does affect test outcome. This, as well as the panel composition and interlot reproducibility, are areas we believe need to be addressed. The PRA-STAT system is rapid, does not require viable cells or complement, and can be automated in part. Resolution of the problems identified here and availability of an IgM-specific conjugate should make this test system a valuable tool in histocompatibility testing.

Antibodies↗
PubMed1980

Recognition of a positive MLC reaction after four hours.

EMT indicator cells were incubated for 1 h in the supernatant of a 3 h MLC. Their electrophoretic mobility was then measured by an analytical, carrier-free electrophoresis system. The alteration of their mobility against a medium-standard was calculated in per cent and correlated with the conventional measured MLR-cpm. According to our results the correlation of the two quantities is statistically highly significant. This method could enable recognition of a positive or negative MLC after only 4 h. This fact could be of great importance for histocompatibility testing and transplantation.

Cell Movement↗
PubMed1978

Vertebral ankylosing hyperostosis (Forestier's disease) and HLA antigens in Pima Indians.

Vertebral ankylosing hyperostosis is a common skeletal disorder among Pima Indians. Prevalence rates of approximately 50% have been reported in Pima Indian males aged 44 or older. The HLA-B27 antigen is also found more commonly in Pima Indians than in Caucasians (18% versus 6%). Because of the increased prevalence of both vertebral ankylosing hyperostosis and HLA-B27 in Pima Indians, a possible association between the two was thought to exist. Therefore, histocompatibility testing was done on 44 adult male Pima Indians age 55 or older with classic vertebral ankylosing hyperostosis and 33 age-matched controls. No significant association between any histocompatibility antigen phenotype or haplotype and vertebral ankylosing hyperostosis was found. This disease, therefore, connot be classified with the HLA-B27 associated spondylarthropathies.

Aged↗
PubMed1985

Recognition of a positive MLR within 4 hours using a carrier free electrophoresis system.

Indicator cells--tanned, surface stabilized sheep erythrocytes--were incubated for 1 h in supernatants of 3 h MLCs. Their electrophoretic mobility was measured by an analytical, carrier free electrophoresis system. The change in their mobility compared with an appropriate control was calculated in per cent and correlated with the conventional measured MLR-cpm. The correlation of the two quantities is statistically highly significant (p less than 0.01). Furthermore, the difference of the electrophoretic mobility values of the group of HLA-D-identical and the groups of HLA-D-haploidentical or -different donors is significant beyond the 1% level (p less than 0.0005). Our method enables, therefore recognition of a positive or negative MLC after only 4 h. Typing for HLA-D-determinants seems to be possible. This could be of great importance for histocompatibility testing and organ transplantation.

Carrier Proteins↗
PubMed1986

[Mixed lymphocyte culture as a compatibility test for bone marrow transplantation].

The reactions in mixed lymphocyte culture (MLC) between family members from 35 healthy families and 80 families of potential bone marrow recipients are shown. From the low MLC reactions seen in 128 pairs of HLA identical siblings, the limit of "Negative Reaction in Mixed Lymphocyte Culture" valid for our own laboratory can now be defined. The importance of mixed lymphocyte culture as a histocompatibility test for bone marrow transplantation is discussed.

Bone Marrow Transplantation↗
PubMed1997

Strategies for determining HLA compatibility in related donor bone marrow transplantation.

BACKGROUND: Although HLA identity between donor and recipient is no longer an absolute requirement for bone marrow transplantation, knowledge of the degree of HLA compatibility is necessary for determining the induction and immunosuppression regimen to be used. In cases of related donor transplantation, HLA compatibility may be assessed by defining the HLA phenotypes at the allele level using high-resolution, DNA-based typing methods or by determining the genotypes of the patient and potential donor from the HLA phenotypes, ascertained by low-resolution typing, of their family members. METHODS: We developed an algorithm that can be used to assess the relative costs of these two approaches. We applied population frequencies for HLA-DR alleles to this algorithm to determine at what cost per test ratio for high-resolution:low-resolution testing the costs of the two approaches are equal. RESULTS: In transplants involving a sibling pair who have the same HLA-A, -B, and -DR antigens, these values are 1.16-1.83 for African-Americans and 1.23-1.97 for Caucasians, depending on the relatives available for testing. With a slight increase in the resolution level achieved with DR antigen testing, the range of values becomes 1.10-1.74. We also estimated that the probability that two antigenically identical siblings have identical HLA-DRB1 alleles is >99% for both African-Americans and Caucasians. A review of 615 cases from our transplant program showed that all of 192 pairs of antigenically identical patients and sibling donors were genotypically or allelically identical, indicating that this estimate is valid. CONCLUSIONS: Transplant programs can apply these algorithms to determine the most cost-effective scheme for histocompatibility testing.

Algorithms↗
PubMed1996

Analysis of HLA-DRB1 alleles using PCR-RFLP and PCR-MPH.

In this study we compare the results of HLA-DRB1 genotyping by PCR-RFLP and PCR-MPH. HLA-DR specificities were also performed by LCT. Samples were obtained from 20 Thai patients who were on the waiting list for kidney transplant. DNA was extracted by phenol-chloroform extraction. It was found that the results gave complete agreement with two methods of DNA typing, however, there were 3 discrepancies in assigning serologic DR specificities and DNA subtypes (p = 0.0001) which were due to the cross reactive antibodies and the lack of potent antisera to define proper HLA-DR subtypes by LCT. These PCR techniques can be applied to identify other alleles such as HLA-DPB1 and HLA-DQB1 which will improve the standard histocompatibility testing in the future.

Alleles↗