DNA typing for HLA-DR, and -DP alleles in a Chinese population using the polymerase chain reaction (PCR) and oligonucleotide probes.

We have determined alleles of HLA-DRB1, DRB3, DRB5, DQA1, DQB1, and DPB1 loci in 91 unrelated healthy individuals from North China. Group-specific PCR primers were employed for the analysis of subsets of DR1, DR2, DR4, DRw52, and DPB. With allele-specific probes, 22 DRB1, 8 DQA1, 13 DQB1, and 12 DPB1 alleles were found in this panel.

Immunogenetics of rheumatoid arthritis and juvenile arthritis.

Starting with the historical background and ending with the most recent data obtained by DNA typing, using PCR and oligonucleotide probes, the role of HLA antigens in rheumatoid arthritis (RA) and in several forms of juvenile arthritis (JA) is reviewed. RA is thought to be associated with an epitope of the third hypervariable region of DRB1 which is shared by several alleles including DR4-Dw4, Dw14, Dw15, DR1, DRw14.2, and DRw10.

DNA typing for class II HLA antigens with allele-specific or group-specific amplification. IV. Typing for alleles of the HLA-DR2 group.

In this study we present an approach for the definition of the alleles belonging to the HLA-DR2 group by DNA typing with oligonucleotide probes. Following methodology similar to that we used previously for the definition of other HLA-DR subsets, we have now developed primers for DR2-DRB1 and DR2-DRB5 amplification, and probes for the identification of sequences that distinguish the subtypes of this group of genes. The method used defines all the previously described alleles at both DR2-associated DRB loci.

Rheumatoid arthritis in Israeli Jews: shared sequences in the third hypervariable region of DRB1 alleles are associated with susceptibility.

Rheumatoid arthritis (RA) is known to be associated with class II HLA antigens in most populations, but recent studies in Israeli Jewish patients showed no significant differences in either DR4 or DR1 between patients and controls. In a previous DR4 subset study we found DR4-Dw15 to be associated with susceptibility (RR = 9.2) but this allele occurred in only 12% of the patients.

A variant of HLA-DR4 determines susceptibility to rheumatoid arthritis in a subset of Israeli Jews.

HLA-DR4 is associated with risk for developing rheumatoid arthritis (RA) in most populations. In Israeli Jews, in whom the Dw10 subtype of DR4 predominates, no association of RA with DR4 has been found. The inability to detect an association could be due to the high frequency of DR4-Dw10.

DNA typing for class II HLA antigens with allele-specific or group-specific amplification. V. Typing for subsets of HLA-DR1 and DR'Br'.

Three DRB1 alleles of the DR1 group including DRB1*0101, DRB1*0102, and DRB1*0103 are currently recognized. The first two of these are defined as HLA-DR1 by serologic typing and as either Dw1 or Dw20 by typing with T cells. DRB1*0103, previously called DR'Br' or DR'BON', is not detectable by serology.

Alloantibodies against donor epidermis and early kidney transplant rejection.

We have previously observed that transplant recipient sera with endothelial antibodies are bound to epidermal cells, as shown by immunofluorescence on sections of skin. It was also reported that early kidney failures that occurred despite negative T cell crossmatches were associated with, and could have been predicted by, a crossmatch with donor skin. Ninety patients undergoing kidney transplantation have now been evaluated using this technique.

DNA typing for class II HLA antigens with allele-specific or group-specific amplification. III. Typing for 24 alleles of HLA-DP.

The second exon of HLA-DPB includes five polymorphic segments with extensive sharing of sequences between alleles. In order to facilitate assignment of specificities in heterozygous individuals, we have used group-specific amplification of two nonoverlapping sets of DPB alleles (here called group A and group B) with especially designed primers. Group A and group B polymerase chain reaction products were hybridized with sequence-specific oligonucleotide probes generating easily recognizable patterns which defined 24 distinct HLA-DPB alleles.

HLA antigens and risk for development of pemphigus foliaceus (fogo selvagem) in endemic areas of Brazil.

Endemic pemphigus foliaceus (EPF), is an autoimmune disease associated with production of IgG antibodies against epidermal antigens. We have tested 38 patients and 50 control subjects living in endemic areas to investigate whether HLA genes are associated with host factors that determine whether or not exposed individuals will develop this disease. A variant of HLA-DR1, an antigen common in Blacks (DRB1*0102), was found to be the main susceptibility factor (relative risk = 7.

HLA antigens in juvenile arthritis. Pauciarticular and polyarticular juvenile arthritis are immunogenetically distinct.

Using DNA techniques, we investigated the role of HLA-DR, DQ, and DP alleles in susceptibility to juvenile arthritis (JA). We studied 2 groups of patients with JA having a different disease prognosis and course. The pauciarticular form is usually benign, while the polyarticular disease frequently leads to joint destruction and disability.

HLA-DR alleles with naturally occurring amino acid substitutions and risk for development of rheumatoid arthritis.

To determine the HLA-DR4 subtypes associated with rheumatoid arthritis (RA), we performed amplification of DR4 DRB1 genes by the polymerase chain reaction and dot-blots with oligonucleotide probes. In 52 HLA-DR4+ RA patients, Dw4 was the predominant subtype. This subtype was found in 45 of 52 patients (86.

Analysis of HLA-DQ genotypes and susceptibility in insulin-dependent diabetes mellitus.

There is evidence that certain alleles at the HLA-DQ locus are correlated with susceptibility to insulin-dependent diabetes mellitus (IDDM) and in particular that DQ beta-chain alleles containing aspartic acid at position 57 are protective. The availability of a large group of patients with IDDM enabled us to assess the role of HLA-DQ alleles in susceptibility to the disease in order to confirm and extend recent observations derived from studies of smaller numbers of patients. Using allele-specific oligonucleotide probes and the polymerase chain reaction, we studied 266 unrelated patients with IDDM and 203 unrelated normal subjects for eight HLA-DQ beta-chain alleles.

DNA typing for class II HLA antigens with allele-specific or group-specific amplification. II. Typing for alleles of the DRw52-associated group.

Codons 9-12 of the first domain of DRB1, which encode the amino acid sequence EYST (in single-letter code), served as the basis for the construction of a polymerase chain reaction primer specific for DRB1 of the whole DRw52 group. Using this primer for the 5' end and a primer for a conserved region at the 3' end, we could amplify selectively the DRB1 genes of DRw17-, w18-, w11-, w13-, w14, and w8-positive haplotypes. DRB3 genes of DR3,DR5 and DRw6 were also specifically amplified, separately.

DNA typing for class II HLA antigens with allele-specific or group-specific amplification. I. Typing for subsets of HLA-DR4.

DNA sequences that distinguish the subsets of HLA-DR4 are also found on several other alleles. This makes typing of heterozygotes with oligonucleotide probes quite impractical. We have therefore developed a procedure in which, in a first step, DNA of the genes to be analyzed is amplified selectively, using group-specific primers.

Prediction of early kidney transplant rejection by a crossmatch with donor skin.

Immunofluorescence staining of donor skin with recipient serum was performed in 25 patients undergoing kidney transplantation. Transplants were performed when cytotoxicity T cell crossmatches with long incubation and with antiglobulin enhancement were negative. In 20 patients the skin crossmatch was negative and all had an uneventful course.

Class II human leukocyte antigen genes and T cell receptor polymorphisms in patients with rheumatoid arthritis.

Human leukocyte antigen (HLA) typing was performed in 174 patients with rheumatoid arthritis and 222 white control subjects. Increases in HLA-DR4 and HLA-DR1 were observed as in previous studies. Each of these appeared to be inherited as dominant risk factors.

Allogeneic lymphocyte stimulation by human spleen. Identification of cells that are very effective for presenting class II HLA antigens.

Human spleen cells were fractionated by percoll density gradient centrifugation and by sorting in the FACS with mixtures of fluorescent antibodies against T cells, B cells, monocytes, and Sig-bearing cells. Cells responsible for powerful MLR stimulation were class II HLA antigen-positive and were concentrated in preparations depleted of all the markers listed above. These cells represented 1-2% of the initial spleen cells.

Associations of HLA-DR4 with rheumatoid factor and radiographic severity in rheumatoid arthritis.

Possible associations between HLA-DR4 and laboratory, radiographic, joint count, functional, and demographic measures of clinical status were analyzed in 154 white patients with rheumatoid arthritis. Overall, 65 percent of the patients were HLA-DR4 positive, similar to other series. HLA-DR4 was associated significantly with the presence of rheumatoid factor and more severe radiographic changes.

HLA-DR4 and other genetic markers in rheumatoid arthritis.

In a new series of RA patients the association with HLA-DR4 was again found to be highly significant. Also increased were the DR4 associated antigens DRw53 and DQw3. Hybridization of a DQ-beta probe to Bg1 II-digested DNA showed that a variant of DQw3, characterized by a 4.

Kidney transplant recipients with long incubation-positive antiglobulin-negative T cell crossmatches.

Low-affinity, cold-reactive antibodies easily removed by washing were not detected by the antiglobulin technique but killed T lymphocytes when washing was omitted, incubation was prolonged, and cytotoxic tests were incubated at room temperature or at 4 degrees C. These antibodies were present in approximately 25% of sera from dialysis patients. Only a subset of such sera (22%) reacted with autologous lymphocytes.

High levels of in vitro IgM rheumatoid factor synthesis correlate with HLA-DR4 in normal individuals.

The association between the HLA-DR4 histocompatibility antigen and in vitro synthesis of IgM and IgM rheumatoid factor (IgM-RF) by blood mononuclear cells was investigated in 35 normal subjects. In vitro cultures of T and B cells were stimulated with pokeweed mitogen, and the secreted IgM-RF, tetanus antibody, and total IgM protein were measured by solid-phase radioimmunoassay. In cultures containing unseparated T cells, IgM-RF production in the DR4+ and DR4- subgroups was not significantly different.

HLA-D region epitopes associated with juvenile arthritis. Recognition by alloreactive T cell clones and alloantisera.

The HLA-D region antigens DR5 (w11,w12), DRw6 (w13,w14), DRw8, DRw52, and DQw1 have previously been shown to be increased in frequency in subsets of patients with juvenile arthritis. Since the HLA-D region is complex (composed of at least 3 subregions encoding multiple molecules, each in turn presenting multiple alloantigenic epitopes), we sought to clarify whether one strongly associated factor might explain the previous findings. To search for the pertinent HLA-D region stimulatory epitopes, alloreactive T cells were primed against DR5 and DRw6 haplotypes and cloned by limiting dilution.