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A comparison of flow cytometry and microcytotoxicity for the evaluation of alloimmune therapy in patients with recurrent spontaneous abortions.

PROBLEM: There is substantial data that support the efficacy of paternal leukocyte immunization (PLI) for the treatment of alloimmune mediated miscarriage; however, there is confusion regarding the laboratory test that should be performed to determine levels of maternal anti-paternal leukocyte antibodies (MAPLA). METHOD: Popular methodologies employed include: 1) microcytotoxicity (MCX), 2) mixed lymphocyte culture (MLC), and 3) cell flow cytometry crossmatch (FCXM). Cell flow cytometry crossmatch correlates well with the more difficult MLC assay although the former proves the more sensitive study. This work compares the MCX assays with FCXM. The study group consisted of ten women who had a history of three or more spontaneous abortions (SABs). All ten had very low levels (< 10%) of MAPLA as measured by FCXM. Following PLI all subjects demonstrated elevated levels (> 50%) of MAPLA by FCXM. At 12 weeks gestation, sera were simultaneously measured for MAPLA by MCX and FCXM. RESULTS: Although all ten patients had very high levels of MAPLA by FCXM during pregnancy, five of ten had antibodies to HLA Class I and two of ten had antibodies to HLA Class II paternal antigens by MCX. Furthermore, all patients who were positive by MCX to paternal Class I antigens were also positive to Class I antigens not seen in either parent. Both patients who were positive by MCX to paternal Class II antigens were also positive to maternal Class II antigens. Notable is that all ten women eventually delivered healthy infants. CONCLUSION: Based on this preliminary study, the MCX assay is neither sensitive or reliable enough to determine the need and/or to monitor the effectiveness of PLI. Flow cytometry should be the modality of choice when determining the need for alloimmunotherapy and to monitor the effectiveness of treatment.

Abortion, Habitual↗

Immunogenetic analysis of successful and rejected bone marrow grafts within one family.

We report the case of an Indonesian patient who required urgent bone marrow transplantation for acute leukaemia and who received successive transplants from two siblings. The first transplant failed while the second was successful. There were some uncertainties in serological typing due to the presence of cross-reacting HLA-B alleles, lack of paternal typing and the use of Caucasoid sera for Indonesian patients. Distinction between the two donors was also difficult. Interestingly, the use of a new DNA technique identified the presence of differences between the patient and the first unsuccessful donor but not the second successful donor.

Adult↗

The association between HLA B5 and ocular involvement in Behçet's disease in southern Turkey.

HLA B5 plays an important role in the aetiology of Behçet's disease. The frequency of HLA B5 differs between racial groups and geographical regions. Ocular involvement, which could lead to serious complications, is a very important feature of Behçet's disease. Various racial groups having different genetic properties live in the Cukurova region, located in southern Turkey. To investigate the role of HLA B5 in Behçet's disease in Cukurova region, HLA B5 antigens were investigated by lymphocytotoxicity assay in 65 patients and 200 healthy controls. A significantly higher frequency was found in the patient group. Neither ocular involvement, nor the type of uveitis showed any correlation with HLA B5, but it was found that patients presenting with frequent ocular attacks had significantly higher HLA B5 positivity than the patients with rare attacks. This observation suggests that HLA B5 would be a negative factor in visual prognosis in Behçet's disease.

Adolescent↗

Lack of association between the MHC linked OTF3 gene and systemic lupus erythematosus.

The objective of this study was to investigate whether an octamer transcription factor gene (OTF3) located within the MHC region of chromosome 6 is involved in determining susceptibility to systemic lupus erythematosus (SLE) in a Spanish population. An OTF3 HindIII polymorphism was characterized by restriction fragment length polymorphism analysis of polymerase chain reaction amplified genomic DNA in 69 patients with SLE and 60 controls. No differences in the OTF3 allelic or genotypic distribution between healthy controls and patients with SLE were found. In the group of patients with diffuse proliferative glomerulonephritis, where we have previously detected the strongest MHC association, we did not observe OTF3 linkage either. In conclusion, the OTF3 gene does not appear to be associated with SLE in the Spanish population. This might be due to the distance of the gene from the HLA class II-III region where more relevant autoimmune-related genes are located.

DNA-Binding Proteins↗

Transfusion-associated graft-vs-host disease. A fatal case caused by blood from an unrelated HLA homozygous donor.

Transfusion-associated graft-vs-host disease (TA-GVHD) is a rare complication of transfusion. We report fatal TA-GVHD in a 63-year-old coronary artery bypass patient of European descent after an RBC transfusion from an unrelated donor. The patient had mild lymphocytopenia and received 2 80-mg doses of methylprednisolone and 7 units of RBCs. On day 14 after the transfusion, he had fever, elevated liver enzyme levels, and a macular rash. Pancytopenia and bone marrow aplasia developed. On day 26, he had a massive gastrointestinal hemorrhage and died. At autopsy, histopathologic findings of the skin, liver, bone marrow, and gastrointestinal tract were consistent with TA-GVHD. One donor of the transfused RBCs (3 days old at transfusion) had a 1-way HLA match with the patient. A method using multiplex polymerase chain reaction is presented. This patient with TA-GVHD and mild immune suppression suggests that blood component irradiation guidelines may need to be reevaluated.

Bone Marrow↗

Stress renders T cell blasts sensitive to killing by activated syngeneic NK cells.

Exposure of primary T cell blasts to stress in the forms of heat, hydrogen peroxide, or high-density growth conditions resulted in a state of enhanced susceptibility to killing by syngeneic IL-2-activated NK cells or lymphokine-activated killer cells, but not to killing by CTL. Cytotoxicity was perforin mediated and was not due to decreased target expression of total MHC class I. The levels of stress used had little effect on cell viability. For thermal stress, sensitization increased with temperature, required a minimum exposure time, and disappeared when cells were given a long enough recovery time. Our data support a model that predicts that activated NK cells play a role in the immunosurveillance of nontransformed stressed cells in normal animals.

Animals↗

[Living donors: immunologic factors].

Transplantations using grafts from living donors were performed on 70 patients with chronic kidney failure, 66 of them involved matching recipients-donors and four involved non-matching recipients-donors. Immunological data were analyzed in 56 pairs of recipients and patients. Of these pairs, one was identical, seven had three identical antigens, 46 were haploidentical at A and B loci, one pair was identical in one antigen and one pair was completely incompatible. The survival of transplanted kidneys largely depended on the degree of histocompatibility. In 33 (59%) transplantations kidneys are functioning from more than 36 months. In the group of seven transplanted pairs with three identical antigens kidneys are functioning in six cases, with four of them functioning from more than 72 months. In the remaining patients (41 patients [73%]) kidneys are functioning, with 8 of them functioning from more than 10 years. The existence of HLA antibodies was investigated. Preimmunization was found in 18 (32%) patients and correlated with the number of blood transfusions. Rejection crises were observed in 12 (21%) patients. As the number of blood transfusions per patient increased the number of rejection crises decreased. Rejection crises were also observed in haploidentical pairs, with a relative risk > 30%. They occurred in the first 2 weeks following transplantation.

Erythrocyte Transfusion↗

NCBI genetic resources supporting immunogenetic research.

The NCBI creates and maintains a set of integrated bibliographic, sequence, map, structure and other database resources to promote the efficient retrieval of information and the discovery of novel relationships. The connections made between elements of these resources permit researchers to start a search from a wide spectrum of entry points. These multiple dimensions of data can be roughly categorized by primary content as text or bibliographic (PubMed, PubMedCentral, OMIM, LocusLink), sequence (GenBank, Reference Sequence Project (RefSeq), dbSNP, MMDB), protein structure (MMDB) or map position (MapView). They can also becategorized by level of expert curation, which may range from validation of submissions from external groups (GenBank, PubMed, PubMedCentral,), to automatic computation (HomoloGene, UniGene), and to highly reviewed and corrected (LocusLink, MMDB, OMIM, RefSeq). Searches can be made by words (in an article title, key words, sequence annotation, database value, author) by sequence (BLAST or e-PCR against multiple sequence databases), or by map coordinates. By computing or curating bi-directional links between related objects, NCBI can represent content on the genetics, molecular biology, and clinical considerations of interest to immunogeneticists. There is also an emerging resource developed by the NCBI in collaboration with the IHWG devoted to the presentation of MHC data (dbMHC). How dbMHC will augment existing resources at the NCBI is described.

Computational Biology↗

[Experimental study on NK cells promoting donor marrow engraftment and hematopoietic reconstitution after MHC haploidentical BMT in mice].

OBJECTIVE: To explore the effect of donor-derived NK cells added to pretreatment conditioning regimen on hematopoietic reconstitution after MHC haplotype-mismatched BMT in mice. METHODS: Murine model of MHC haplotype-mismatched BMT was established by using BALB/c(H-2d) x C57BL/6(H-2b) (CB6F(1)(H-2d/b)) mouse as recipient, and C57BL/6(H-2b) mouse as donor. Fifty recipient mice were divided into 5 groups. The mice in the first three groups were each infused 1 x 10(6), 5 x 10(5), 2 x 10(5)/mouse donor-derived NK cells, respectively before TBI ((60)Co, 9.0 Gy) and then conditioned with TBI, followed by infusion of C57BL/6(H-2b) mice bone marrow cells four hours later. The mice in the fourth group received TBI only, and in the fifth group, TBI and BMT at the some doses as the first three groups. Hematopoietic reconstitution, survival time, body weight, histopathology of the recipients were followed up. RESULTS: (1) Survival time was (5.15 +/- 0.66) days for the fourth group, and > 30 days for the other 4 groups. (2) Leukocyte and platelet counts at day 10 after BMT were (0.99 +/- 0.22) x 10(9)/L and (61.0 +/- 7.27) x 10(9)/L respectively for the fifth group and (2.01 +/- 0.21) x 10(9)/L, (101.50 +/- 16.34) x 10(9)/L; (1.98 +/- 0.29) x 10(9)/L, (99.50 +/- 16.41) x 10(9)/L and (1.97 +/- 0.21) x 10(9)/L, (98.0 +/- 16.19) x 10(9)/L for the first three groups, respectively. Histopathology displayed no GVHD in all the groups. CONCLUSION: Donor-derived NK cells could promote hematopoietic reconstitution after MHC haplotype-mismatched BMT in mice.

Animals↗