ABSTRACT
【Objective】 To statistically analyze the characteristics of ambiguous results of HLA-DPB1 genotyping given by AllType NGS 11-loci sequencing reagent via two next generation sequencing platforms, i. e. Ion Torrent S5 and Illumina Miseq. 【Methods】 A total of 434 samples from patients or donors were genotyped for HLA-DPB1 locus using AllType NGS 11-loci sequencing reagent from One Lambda company; 336 samples of them were sequenced via the Ion Torrent S5 platform and other 98 samples were sequenced via the Illumina Miseq platform. All 434 samples were genotyped for HLA-DPB1 gene simultaneously using PCR-SSO flow fluorescent bead method. The ambiguous genotypes of HLA-DPB1*13∶01∶01/107∶01 were distinguished by Sanger sequencing. The HLA-DPB1 genotype results by NGS method were assigned by TypeStream Visual professional software, and the ratio of ambiguous combination was calculated by direct count method. 【Results】 Ambiguous results were found in 357 out of 434 samples, accounting for 82.3% (357/434) when HLA-DPB1 allele was assigned to the third field using NGS method. Ambiguous results with 45 types were given in 275 out of 336 samples by the Ion Torrent S5 platform, accounting for 81.8% (275/336) and 82(with 27 types) out of 98 samples by the Illumina Miseq platform, accounting for 83.7% (82/98). All samples were re-genotyped for HLA-DPB1 gene by PCR-SSO, and none HLA-DPB1 allele had been missed by NGS. A total of 43 ambiguous alleles in HLA-DPB1*13∶01∶01/107∶01 involving 41 samples were distinguished by Sanger sequencing; HLA-DPB1*13∶01∶01 were detected in 25 (58.1%, 25/43) and HLA-DPB1*107∶01 in 18 (41.9%, 18/43). 【Conclusion】 There were still a high proportion of HLA-DPB1 ambiguous combinations using the AllType NGS 11-loci sequencing reagent. Sequencing exon 1 of HLA-DPB1 gene by Sanger sequencing can resolve part of the ambiguous results in HLA-DPB1*13∶01∶01/107∶01 alleles.
ABSTRACT
ABSTRACT Background: The impact of HLA-DPB1 compatibility and its role as a transplantation antigen in haploidentical-related hematopoietic stem cell transplant (haplo-R-HSCT) have not been established, and a negative effect on survival has been suggested. Objective: The objective of the determine was to study the frequency and clinical effects of incompatibility at the HLA-DPB1 locus in the haplo-R-HSCT setting. Methods: Clinical records and electronic files of 91 patients with a hematological disease who underwent haplo-HSCT from January 2009 to October 2017 in a university medical center were scrutinized. Overall survival (OS) was estimated by the Kaplan-Meier method; the cumulative incidence of transplant-related mortality (TRM) and relapse rates was determined. Acute graft-versus-host disease was assessed by binary logistic regression. Cox regression model with a 95% confidence interval was used to examine the association between the different variables and their effect on OS. Results: Of the 91 donor-recipient pairs, 24 (26.37%) shared complete DPB1 identity, 60 (65.93%) had a mismatch at one allele, and 7 (7.70%) were mismatched at two alleles. Twenty-four different HLA-DPB1 alleles were found; the most frequent were DPB1*04:01 (34.1%) and DPB1*04:02 (27.5%). Two-year OS, the cumulative incidence of TRM and relapse was 51.3 ± 6.8%, 28 ± 6% and 60 ± 7.8% for all haplo-related transplants, respectively, with no statistical difference between HLA-DPB1 matched and partially matched patients. In Cox regression analysis, no risk factors associated with OS, TRM, or relapses were identified. Conclusion: HLA-DPB1 mismatching in the haplo-R-HSCT setting did not influence transplant outcomes and was clinically tolerable. A high degree of homozygosity was found.
Subject(s)
Humans , Male , Female , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Young Adult , Hematopoietic Stem Cell Transplantation/methods , HLA-DP beta-Chains , Transplantation, Haploidentical , Hematologic Diseases/surgery , Survival Rate , Retrospective Studies , Treatment Outcome , Patient Selection , Donor Selection , Hematologic Diseases/mortalityABSTRACT
Objective:In this study,we examined the HLA-DPB1 alleles in patients with tuberculosis,and health individuals attempt to investigate the association between the polymorphism of HLA-DPB1 gene and pulmonary tuberculosis in Han population in Shihezi area in Xinjiang uygur autonomous region of China.Methods:High-resolution typing of DPB1 was performed by the sequence-based typing ( SBT) method using the SBT-HLA-DPB1 generic DNA typing kit.Results:In the controls ,17 HLA-DPB1 alleles were ob-served,the HLA DPB1*0501 (28.1%) and HLA DPB1*0201(27.6%) frequency was significantly higher than other sites ,the first and second respectively.The frequency of HLA-DPB1* 0201 was observed significantly increased in patient group compared with control group ( P<0.05 );the frequency of HLA-DPB1* 0501 was observed significantly decreased in patient group compared with control group ( P<0.05 ).Conclusion: The gene frequencies of HLA-DPB1 in Xinjiang Shihezi Han population are roughly in consistence with other northern Chinese Han population;the HLA-DPB1*0201 may be the protective factor to pulmonary tuberculosis , and HLA-DPB1*0501 may be susceptible to pulmonary tuberculosis in Han population from the Xinjiang uygur autonomous region of China.
ABSTRACT
Objective:To study the HLA-DPB1 allele polymorphism in Ewenki from Inner Mongolian.Methods:HLA-DPB1 allele polymorphism in normal Ewenki were determined by PCR with sequencing-based-typing(SBT).Results:20 HLA-DPB1 alleles were observed and compared with other ethnic groups,the allele frequency of HLA-DPB1*02012(24.4%) and DPB1*0402(22.6%),DPB1*0401(20.2%),DPB1*0501(10.7%) are highest,while others are lower.Conclusion:The distributions of HLA-DPB1 alleles frequencies in normal Ewenki from Inner Mongolia has a unique style.It is most important to further study anthropology and related to illness in Ewenki nationality.