Abstract
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. We also established a routine procedure for distinguishing HLA-DP homozygosity from failed amplification in one of the alleles. Our results showed that when only one allele was detected, failure of amplification had occurrured in less than 4% of the cases. DNA typing with this method correlated well with primed-lymphocyte typing for HLA-DP in the Tenth Workshop, as determined by us in assays performed on the workshop B-cell lines. Two normal panels of unrelated subjects were tested to obtain population frequencies. We conclude that this method is simple, relatively quick, and accurate. It is the method of choice for studies to determine the role of HLA-DP alleles in T cell reactions, in various diseases, and in transplantation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 60-68 |
| Number of pages | 9 |
| Journal | Human Immunology |
| Volume | 30 |
| Issue number | 1 |
| DOIs | |
| State | Published - 1991 |
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ASJC Scopus subject areas
- Immunology
- Immunology and Allergy
Cite this
DNA typing for class II HLA antigens with allele-specific or group-specific amplification III. Typing for 24 alleles of HLA-DP. / Fernandez-Vina, Marcelo; Moraes, M. Elisa; Stastny, Peter.
In: Human Immunology, Vol. 30, No. 1, 1991, p. 60-68.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - DNA typing for class II HLA antigens with allele-specific or group-specific amplification III. Typing for 24 alleles of HLA-DP
AU - Fernandez-Vina, Marcelo
AU - Moraes, M. Elisa
AU - Stastny, Peter
PY - 1991
Y1 - 1991
N2 - 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. We also established a routine procedure for distinguishing HLA-DP homozygosity from failed amplification in one of the alleles. Our results showed that when only one allele was detected, failure of amplification had occurrured in less than 4% of the cases. DNA typing with this method correlated well with primed-lymphocyte typing for HLA-DP in the Tenth Workshop, as determined by us in assays performed on the workshop B-cell lines. Two normal panels of unrelated subjects were tested to obtain population frequencies. We conclude that this method is simple, relatively quick, and accurate. It is the method of choice for studies to determine the role of HLA-DP alleles in T cell reactions, in various diseases, and in transplantation.
AB - 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. We also established a routine procedure for distinguishing HLA-DP homozygosity from failed amplification in one of the alleles. Our results showed that when only one allele was detected, failure of amplification had occurrured in less than 4% of the cases. DNA typing with this method correlated well with primed-lymphocyte typing for HLA-DP in the Tenth Workshop, as determined by us in assays performed on the workshop B-cell lines. Two normal panels of unrelated subjects were tested to obtain population frequencies. We conclude that this method is simple, relatively quick, and accurate. It is the method of choice for studies to determine the role of HLA-DP alleles in T cell reactions, in various diseases, and in transplantation.
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U2 - 10.1016/0198-8859(91)90072-H
DO - 10.1016/0198-8859(91)90072-H
M3 - Article
C2 - 2001976
AN - SCOPUS:0026088001
VL - 30
SP - 60
EP - 68
JO - Human Immunology
JF - Human Immunology
SN - 0198-8859
IS - 1
ER -