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PubMed1976

Lymphocytotoxic antibody detection and crossmatching for renal transplantation.

Sera of 134 recipients awaiting first renal transplants have been tested for cytotoxic antibody by three methods in order to assess the different techniques, and various factors in the techniques have been investigated. The use of only one technique would have failed to detect some of the antibodies found in the sera of 35 patients and it was necessary to use both a 37 C test and the NIH 20 C technique in order to demonstrate them. Results on 402 serial samples from 108 patients indicate the importance, because of fluctuation in antibody levels. of using more than one sample, i.e., previous sample in addition to a fresh specimen from any patient when crossmatching prior to renal transplantation. Lymphocyte suspensions from 44 of 118 cadaver donors were found to be unsatisfactory for accurate cytotoxic crossmatching due to a high background of dead cells. DNase renders such suspensions suitable for use and should be used in such cases. It is felt that if more than one technique were generally used, improved detection of cytotoxic activity would result and would help to resolve present conflicting opinions about the effects of cytotoxic antibody on graft survival. Results are given of 19 patients who had formed cytotoxic antibody prior to renal transplantation in the Unit. Eighteen of these have surviving grafts.

Antibodies↗
PubMed1976

Recipient selection for renal retransplantation.

Analysis of over 1,000 kidney retransplants revealed an important effect of first graft duration and presensitization on graft outcome. Second cadaver donor grafts into patients without preformed cytotoxic antibodies and a first graft duration of more than 12 months survived at a high 1-year rate of 71 +/- 6%, in contrast to 29 +/- 6% in those whose first graft had failed in 1-3 months (P is less than 0.0001). Third or fourth transplants into patients with preformed cytotoxins survived at the poor rate of 17 +/- 6% at 1 year. To repeat all HLA mismatches from the first graft was found to result in decreased second graft survival.

Cadaver↗
PubMed1979

Do blood transfusions enhance the possibility of a compatible transplant?

Blood transfusions prior to first cadaver kidney transplants have a significant beneficial effect on graft survival and, in this sense, appear to enhance the possibility of a compatible transplant. This desirable effect, however, occurs concomitantly with an increased degree of sensitization, which in turn reduces the likelihood of identifying a compatible kidney by direct crossmatch testing. This report illustrates that the beneficial effect is achieved with one to five transfusions prior to transplantation, but that more transfusions afford no additional benefits. In addition, the presence of cytotoxic antibodies per se does not have an adverse influence on graft survival. Liberal transfusion policies are therefore indicated in cadaver transplant candidates, but more than five transfusions prior to transplantation should probably be avoided unless clinically necessary.

Antibodies↗
PubMed1980

HLA-DR typing by radioimmunoassay.

A radioimmunoassay procedure is described by which peripheral blood lymphocytes can be typed for HLA-DR specificities. The major advantages of this method are the following: simple and reproducible procedure, no need for B lymphocyte separation, no need for optimal viability, and no need for preabsorption of antisera with platelets. This method will find an application in the genetic and biochemical analysis of the HLA complex, and in the clinical tests of Ia antigens for diagnostic or prognostic purposes and in retrospective transplant studies.

Antibody Specificity↗
PubMed1980

Major histocompatibility complex matching and other factors influencing skin allograft survival in related and unrelated rhesus monkeys.

Skin allografting was performed in rhesus monkeys to study the influence of matching for products of the RhLA region and of various other parameters. As expected, the longest mean survival times (MSTs) were observed when donors were RhLA-identical siblings; the MST seemed a bit shorter when grafts from at least two RhLA-nonidentical siblings were simultaneously present. Matching for RhLA-A and B locus antigens was studied in RhLA-haploidentical and in unrelated combinations; in both categories, the MST was about 12 days if there were no A or B antigen disparities, 10 days if there was one A or B disparity, and about 9 days for two or more disparities. A positive effect of matching for D/DR antigens and mixed lymphocyte culture (MLC) nonresponsiveness between host and donor was questionable; additional data are required to confirm the minor beneficial effect observed. With regard to the sex of donors and recipients, female recipients rejected skin allografts more rapidly than did males in related and unrelated combinations. In this respect, females with a known or possible history of pregnancy were not different from nulliparous animals. A possible influence of donor sex on skin allograft survival remained equivocal. The i.v. administration of donor-specific antigen before skin grafting led to a significant prolongation of the MST; in the majority of combinations with prolonged graft survival, B cell-specific antibodies were demonstrable in the recipients' sera. An "enhancing" influence of such antibodies is possible, although the available data do not prove such an effect.

Animals↗
PubMed1986

Differences in immunoglobulin subclasses between dye exclusion and 51Cr release complement-dependent lymphocytotoxicity assays.

Paradoxical differences previously noted between lymphocytotoxicity detected by dye exclusion at room temperature (CDCE) or by 51Cr release (CDC51Cr) at 37 degrees C in maternal antipaternal complement-dependent lymphocytotoxicity have suggested that CDCE and CDC51Cr at 37 degrees C, but not at 20 degrees C, may detect different immunological antibody-antigen interactions. Reactions in the two test systems against the same target cells were compared in sera from known immune dialysis patients, secondary aborting women, and refractory platelet recipients before and after heat treatment of sera, absorption with solid-phase heparin, anti-light-chain augmentation, and the addition of murine monoclonal anti-IgG subclass antibodies. The results demonstrate significant differences between the two tests using the same target and sera. Further, the results imply the presence of an inhibitor and an inhibitor of inhibitor in sera. The involvement of different immunoglobulin subclasses was shown in the two tests. These data demonstrate the necessity for further study of the nature of the differences in the mechanisms of these clinically important antibody-detecting systems.

Abortion, Spontaneous↗
PubMed1991

Renal transplantation despite a positive antiglobulin crossmatch with and without prophylactic OKT3.

The antiglobulin crossmatch (AGXM) is a sensitive technique employed by many transplant centers to enhance detection of preformed antibody to donor antigens that may cause hyperacute rejection. However, positive AGXM may detect irrelevant or very low titers of anti-HLA antibody precluding transplantation in suitable recipients. To investigate the significance of a positive AGXM, cadaveric renal transplantation was carried out despite a weakly positive AGXM (defined as cell killing above background but not greater than 20%) in 48 recipients. In an initial group (n = 10), maintained on triple therapy (cyclosporine, azathioprine, and prednisone), accelerated acute rejection occurred in 4 recipients and 3 grafts were lost. A subsequent group (n = 38) was treated with a prophylactic course of OKT3 then triple therapy. There were no episodes of accelerated acute rejection (P less than 0.01) although clinical hyperacute rejection claimed one graft and the incidence of delayed graft function was high (75%). The prophylactic OKT3 group had a reduced incidence of acute rejection (0.5 versus 1.0) per recipient and the onset of first episodes was delayed (mean onset: 13 versus 35 days after transplantation). One year actuarial primary graft survival was 88% in the prophylactic OKT3 group as compared with only 50% in the initial group. The outcome in the positive AGXM group was similar to a concurrent group (n = 32) with a negative AGXM and immediate graft function. On the other hand, the subset of positive AGXM regraft recipients treated with prophylactic OKT3 fared poorly, with a 36% (4/11) incidence of primary nonfunction. In summary, a positive AGXM, as defined in this report, is not a contraindication to primary renal transplantation--in fact, the use of the AGXM will identify recipients that would benefit from prophylactic OKT3.

Adult↗
PubMed2003

Spleen transplantation in miniature swine: surgical technique and results in major histocompatibility complex-matched donor and recipient pairs.

BACKGROUND: Spleen transplantation (Tx) between some strains of rodents can lead to donor-specific tolerance either spontaneously or after a short course of immunosuppression. This study developed a surgical technique for spleen Tx in miniature swine to investigate its immunologic impact in a large animal model. METHODS: The preferred surgical technique of spleen Tx (n=8) involved excision of the donor spleen with its vascular pedicle to the aorta and portal vein. Carrel patches of donor aorta and portal vein were anastomosed to the abdominal aorta and inferior vena cava, respectively, of the (splenectomized) recipient. The results in four major histocompatibility complex-matched pairs that were mismatched for the porcine allelic antigen are reported. Two recipients were untreated, one received a 12-day course of cyclosporine A (CsA) alone, and one received thymic irradiation (700 cGy) and CsA. Hematopoietic cell chimerism was followed by fluorescence-activated cell sorter, and graft survival was assessed by histology. RESULTS: Spleen Tx was technically successful. In two untreated pigs, chimerism was detected in the blood (maximum 5% for 17 and 25 days) and lymph nodes (maximum 6% for 28 and 56 days), but both grafts showed histologic rejection by day 28. In two treated pigs, chimerism was present in the blood for 47 and 57 days, and rejection was prevented, with follow-up for 57 and 217 days, respectively. CONCLUSION: Spleen Tx in major histocompatibility complex-matched pairs treated with CsA+/-thymic irradiation results in prolonged chimerism and is associated with the development of in vivo unresponsiveness to the transplanted spleen.

Animals↗
PubMed1977

HLA-D-associated 'Ia-like' antigens on human macrophages.

Human macrophages procured by culture of adherent peripheral blood mononuclear cells were investigated for reactivity in complement-dependent cytotoxicity tests with antisera having specificity for HLA-D-associated 'Ia-like' B-cell antigens. The antisera were found to be cytotoxic for 30%-70% of the macrophages from donors whose B cells were also reactive, and absorption with B cells also removed macrophage cytotoxicity. A B-cell xenoantiserum was also cytotoxic to macrophages. It is concluded that the HLA-D-associated 'Ia-like' antigens present on B cells are also expressed by macrophages, at least on a subpopulation.

B-Lymphocytes↗
PubMed1977

Selection of haploid spermatozoa and its application to HLA-D typing.

The expression of HLA-A, -B, and -D determinants on the surface of spermatozoa is haploid. Using cytotoxicity and appropriate HLA antisera and complement, we were able to select haploid spermatozoa. The surviving sperms behaved similarly to homozygous lymphoid cells when cultured in mixed sperm lymphocyte culture (SLC). This method is being applied to define new HLA-D specificities, to improve typing and matching for bone marrow transplantation and kidney transplantation and to study their association with various diseases.

Complement System Proteins↗
PubMed1992

Neonatal alloimmune thrombocytopenia due to anti-HPA-5b (Br(a), Zav(a), Hca): the importance of third-generation platelet antibody detection techniques, a case report.

Investigation of the maternal serum in a case of suspected alloimmune neonatal thrombocytopenia by conventional, second-generation platelet serological assays (platelet radioactive antiglobulin test [PRAT], platelet suspension immunofluorescence test [PSIFT] and solid-phase adherence assay (SPAA, 'Capture-P') demonstrated only the presence of HLA class-I antibodies of limited specificities: no platelet-specific antibodies were detectable. The use of a third generation, glycoprotein capture assay (monoclonal antibody-specific immobilization of platelet antigens, MAIPA) revealed the additional presence of anti-HPA-5b with a titre of 1 in 32. Despite this relatively high titre, and the fact that it was able to induce a prolonged thrombocytopenia, this antibody was not detectable by conventional assays. In view of these findings we conclude that the use of MAIPA is essential when investigating cases of suspected alloimmune neonatal thrombocytopenia.

Abnormalities, Multiple↗
PubMed1985

Disease risk estimates from marker association data. Application to individuals at risk for hemochromatosis.

The recessive hemochromatosis gene is both linked to the HLA region on chromosome 6 and nonrandomly associated with certain HLA alleles. The use of linked HLA markers to trace known hemochromatosis genes within a family is well known, but using the population associations to detect unsuspected disease genes has not been fully appreciated. Thus, while HLA typing has been utilized to detect asymptomatic affected siblings, it has not been applied to other relatives. We propose a method in which Bayes' rule is used to calculate the probability that designated HLA marker haplotypes, brought into the family by spouses, have attendant hemochromatosis genes. The A3, B14 and A3, B7 haplotypes are such high risk markers. When these haplotypes are inherited from the unaffected parent, the offspring of an individual with hemochromatosis is at marked increased risk for the disease. When A3 and B14 are absent from the HLA marker haplotype, however, the risk of having a hemochromatosis gene is less than that for the general population. This approach should be helpful in identifying family members at a higher risk for developing the disease and who may then undergo appropriate periodic screening.

Bayes Theorem↗