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Molecular analysis of HLA genes for the selection of unrelated bone marrow donor.

The selection of fully matched unrelated volunteer donors (UVD) in BMT requires a molecular characterization of MHC polymorphism, since most phenotypically HLA-identical donors can be non-identical when analyzed at a genomic level. The present report describes a molecular typing protocol for HLA genes developed for the selection of UVD, and its application to some donor-recipient pairs. The protocol involves three successive steps. Firstly, PCR with sequence-specific primers for HLA-DRB1 and -DQB1 genes is performed to identify the major alleles of the recipient. PCR-fingerprint matching is then introduced for HLA-A, B, C and DRB, DQB and DPB genes to screen prospective donors. Those showing matched fingerprinting patterns are finally submitted to direct sequencing of the DRB1 gene. DPB compatibility is assessed by oligotyping when there are several potential class I and DRB matched donors. This strategy was applied retrospectively to three BMT recipients and their previously selected donors. Three other patients and their 12 prospective donors were submitted to our protocol before BMT. Clinical evaluation of transplant outcomes indicates the primary importance of complete DRB and class I matching, while DQB and DPB compatibility seems to be less critical.

Base Sequence↗

The importance of HLA genes to susceptibility in the development of juvenile diabetes mellitus. A study of 93 patients and 68 first degree blood relations.

The importance of genetically determined antigens of the HLA system in etiology and pathogenesis of juvenile onset diabetes (JOD) was studied in 93 JOD-patients and 68 blood relations. A close association was found between JOD and B-locus antigens B8 and Bw15, and C-locus antigen Cw3. Patients positive for one of these antigens have a 2-3 times -- and those positive for both B8 and Bw15 -- a 8.6 times greater chance of developing JOD, Evidence for a genetic heterogeneity between childhood type and later onset JOD could be obtained. B8 seems to play a particularly important role in childhood-type diabetes, whereas in the later onset JOD the antigens Cw3 and/or Bw15 might possibly represent an additional predisposing factor. Family studies have revealed a close correlation between glucose intolerance and those genes associated with JOD in blood relations below age 35. HLA-B7 which could be detected in that group of blood relations, and which was found statistically decreased in JODs might even exert some protective role. Incidence of haplotye identity in glucose intolerant siblings was almost three times higher than expected. These data provide evidence for the existence of a genetic basis which determines the susceptibility to develop JOD.

Adolescent↗

Genetics of histocompatibility.

Molecular studies of the major histocompatibility complex region, its genes, and its products are progressing in a number of different directions, and this progress is becoming of increasing interest not only to immunologists but also to geneticists and clinicians. Here I select only certain aspects of this field, stressing the biological importance of a few recent developments. This review concentrates on two topics of clinical importance and where important recent progress is reported: the "revolution" of HLA genotyping for histocompatibility matching in transplantation and the elucidation of a genetic disease of gene regulation, primary major histocompatibility complex class II deficiency (or bare lymphocyte syndrome).

Animals↗

[HL-A system and diabetes mellitus].

In 19 families of juvenile diabetes patients intravenous glucose tolerance was tested and HLA antigens were determined. A total of 68 first degree blood relations (siblings, parents, children) was studied. Taking the age dependent variabilities of the glucose assimilation coefficient (k-value) into consideration, glucose intolerance was found in 35.5% of the blood relations. Particularly in blood relations (above all in siblings) aged under 35 and with glucose intolerance, a trend to increased frequencies of those HLA antigens (B8, BW15, CW3) associated with juvenile diabetes was found, but it is not yet clear whether this association will be of practical significance.

Adult↗

Comparison of haplotypes of the major histocompatibility complex in the rat. I. The Ag-B7 (H-1g) and Ag-B8 (H-1k) haplotypes.

Two haplotypes of the major histocompatibility complex of the rat, Ag-B7 and Ag-B8, have been compared with known H-1 haplotypes using the F1 skin-graft test and the dextran haemagglutination test. Both of these Ag-B haplotypes were different from the known H-1 haplotypes and determined different private specificities. The Ag-B7 haplotype was denoted as H-1g and the Ag-B8 haplotype as H-1k. The complex structure of the serologically detected antigenic products of these haplotypes was determined by means of H-1 congenic lines.

Alleles↗

Histocompatibility-2 (H-2) polymorphism in wild mice.

Red blood cells of 40 wild mice captured at four different locations in the Ann Arbor area were typed for the presence of 15 different H-2 antigens by direct hemagglutination test. Fifteen different phenotypes were found and all were different from those determined by known H-2 alleles of the laboratory mice. Great similarity between phenotypes of mice from the sane location was observed. This is interpreted as further evidence for the deme structure of natural mouse population.

Animals↗

Deletion of the HLA-B Gene in One of the Inherited Haplotypes in a Northern European Family.

Targeted next generation sequencing-based HLA typing of a 17-year-old female transplant patient showed homozygosity for the HLA-B allele. The segregation analysis of HLA haplotypes of family members only allowed the conclusion that the B-allele was deleted in the haplotype inherited from the father and accordingly paternal grandfather, resulting in false homozygous genotyping. The subsequent whole-genome sequencing of the patient and her father confirmed an approximately 85 kb deletion at 6p21.33 from the 5' end of the HLA-B to the 3' end of the HLA-C gene extending telomeric to HLA-C.

Adolescent↗

The major histocompatibility complex and its relationship to allergic disease.

Two tests, the mixed leukocyte culture (MLC) and cell-mediated lympholysis (CML) tests, have been used as in vitro models of the in vivo allograft reaction. These tests have been applied to histocompatibility testing for transplantation as well as to assay of immune function. They involve the use of peripheral blood lymphocytes as "responding" cells in mixed leukocyte culture or as "stimulating" cells in the generation of a proliferative or a cytotoxic response. The proliferative events in MLC are primarily in response to LD antigens of the major histocompatibility complex; the cytotoxic cells use as their targets the SD antigens of that complex. A new method for defining the LD antigens, the primed LD typing (PLT) test, is based on in vitro sensitization of lymphocytes to certain LD antigens of the major histocompatibility complex and their subsequent restimulation with test cells.

ABO Blood-Group System↗

Power of association test for detecting minor histocompatibility gene causing graft-versus-host disease following bone marrow transplantation [correction].

Incompatibility of minor histocompatibility antigen (mHa) is a major cause of acute graft-versus-host disease (GVHD) following bone marrow transplantation in human leukocyte antigen (HLA)-matched donor-recipient pairs. To avoid acute GVHD, as many mHa genes as possible need to be identified. In this study, we introduce a comparison of two proportions as an association test for detecting mHa genes in HLA-matched pairs with and without GVHD. Assuming multiple mHa loci, each with two alleles, we evaluated the effects of (1). minor allele frequency of the mHa locus of interest (denoted by p), and (2). probability of GVHD developing in a donor-recipient pair being incompatible at an mHa locus (denoted by r) on the powers of association tests for unrelated pairs and for sib pairs. Our results showed that based on a candidate gene approach, an mHa gene with high p and r values can be detected by the association test with a small sample size. Application of the present method to the Japanese population revealed that the association test for unrelated pairs is more suitable for detecting an mHa gene with a high r value than that for sib pairs. The present method will be helpful to researchers who evaluate the power of association study in advance.

Bone Marrow Transplantation↗