The characteristic of HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DRB3/4/5, HLA-DQA1, HLA-DQB1, HLA-DPA1, and HLA-DPB1 alleles in Zhejiang Han population.

The Zhejiang Han population, a subgroup of the Southern Han ethnic group, resides in Zhejiang Province, situated on the southeast coast of China. In this study, we conducted HLA genotyping for 813 voluntary umbilical cord blood donors from the Zhejiang Han population, targeting 11 HLA loci, namely HLA-A, HLA-B, HLA-C, HLA-DRB1, HLA-DRB3/4/5, HLA-DQA1, HLA-DQB1, HLA-DPA1, and HLA-DPB1, using the next-generation sequencing method. Our analysis of the alleles and haplotypes revealed a high degree of polymorphism within these loci. A total of 289 unique HLA alleles were identified, with the HLA-B locus exhibiting the most significant diversity, while HLA-DRB4 displayed the lowest variation. Due to the inherent limitations of the sequencing method, some unresolvable alleles in the specific loci, such as HLA-DRB1, HLA-DPA1, and HLA-DPB1, were assigned as G group designation. In our comprehensive analysis across all 11 HLA loci, a total of 1204 haplotypes were estimated. The distribution of these alleles was similar to those of the Chinese Southern Han population while highly different from the Caucasian population. These findings contribute to a deeper understanding of the genetic characteristics of HLA loci within the Chinese Southern Han population.

[Sequence analysis and identification of a novel HLA-DPB1*02:01:69 allele by third-generation sequencing].

OBJETIVE: To analyze the sequence of a novel HLA-DPB1 allele in an individual. METHODS: A individual identified from the database of blood donors for matched platelet transfusion at the Blood Center of Zhejiang Province in May 2022 was selected as the study subject. HLA genotype of the individual was determined by next-generation sequencing (NGS) on an Ion Torrent S5 platform. The sequence of the HLA-DPB1 locus was also determined by NGS on an Illumina Miseq platform and third-generation sequencing using Oxford Nanopore MinION. This study was approved by the Blood Center of Zhejiang Province (Ethics No. 2021-001). RESULTS: A novel HLA-DPB1*02 allele was identified in the specimen, for which the closest genotype was HLA-DPB1*02:new,17:01:01G, with the variant located in exon 3. Meanwhile, the NGS also revealed a novel HLA-DPB1*17 allele, with the closest genotype being HLA-DPB1*02:01,17:new. Both the HLA-DPB1*17:01:01:01 and HLA-DPB1*02 alleles were identified by third-generation sequencing. Compared with the HLA-DPB1*02:01:02:01 allele, the novel allele had a G>A variation at position 369 in the exon 3, which however did not result in amino acid change. CONCLUSION: A novel HLA-DPB1 allele has been identified and validated by both NGS and TGS, which has been named as HLA-DPB1*02:01:69 by the World Health Organization Committee on Nomenclature of Factors of the HLA System.

The distributions of HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 allele and haplotype at high-resolution level in Zhejiang Han population of China.

The distributions of HLA allele and haplotype are variable in different ethnic populations and the data for some populations have been published. However, the data on HLA-C and HLA-DQB1 loci and the haplotype of HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 loci at a high-resolution level are limited in Zhejiang Han population, China. In this study, the frequencies of the HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 loci and haplotypes were analysed among 3,548 volunteers from the Zhejiang Han population using polymerase chain reaction sequencing-based typing method. Totals of 51 HLA-A, 97 HLA-B, 45 HLA-C, 53 HLA-DRB1 and 27 HLA-DQB1 alleles were observed. The top three frequent alleles of HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 loci were A*11:01 (23.83%), A*24:02 (17.16%), A*02:01 (11.36%); B*40:01 (14.08%), B*46:01 (12.20%), B*58:01 (8.50%); C*07:02 (18.25%), C*01:02:01G (18.15%), C*03:04 (9.88%); DRB1*09:01 (17.52%), DRB1*12:02 (10.57%), DRB1*15:01 (9.70%); DQB1*03:01 (22.63%), DQB1*03:03 (18.26%) and DQB1*06:01 (10.88%), respectively. A total of 141 HLA-A-C-B-DRB1-DQB1 haplotypes with a frequency of ≥0.1% were found and the haplotypes with frequency greater than 3% were A*02:07-C*01:02:01G-B*46:01-DRB1*09:01-DQB1*03:03 (4.20%), A*33:03-C*03:02-B*58:01-DRB1*03:01-DQB1*02:01 (4.15%), A*30:01-C*06:02-B*13:02-DRB1*07:01-DQB1*02:02 (3.20%). The likelihood ratios test for the linkage disequilibrium of two loci haplotypes was revealed that the majority of the pairwise associations were statistically significant. The data presented in this study will be useful for searching unrelated HLA-matched donor, planning donor registry and for anthropology studies in China.

Infection of common marmosets with hepatitis C virus/GB virus-B chimeras.

UNLABELLED: The development of vaccination and novel therapy for hepatitis C virus (HCV) has been hampered by the lack of suitable small-animal models. GB virus B (GBV-B), closely related to HCV, causes viral hepatitis in common marmosets (Callithrix jacchue jacchus) and might represent an attractive surrogate model for HCV infection. However, differences exist between GBV-B and HCV in spite of a short genetic distance between the two viruses. Here we report common marmosets infected with two HCV/GBV-B chimeras containing HCV structural genes coding for either whole core and envelope proteins (CE1E2p7) or full envelope proteins (E1E2p7) substituted for the counterpart elements of GBV-B. Naïve animals intrahepatically injected with chimeric RNA transcripts or intravenously injected with sera from primary infected animals produced high levels of circulating infectious chimeric viruses and they developed chronic infection. Tacrolimus-treated marmosets inoculated with a CE1E2p7 chimera had higher viral loads and long-term persistent infection. A moderate elevation of serum aspartate aminotransferase (AST) levels was observed in parallel with viral replication. Chimeras recovered from liver samples revealed 1/958 adaptive viral mutations. Histopathological changes typical of viral hepatitis were observed in liver tissues from all types of HCV chimeras-infected marmosets. HCV core and E2 proteins were detected in liver tissues from infected animals by immunohistochemical staining. Fluctuations of chimeric virus replication in marmosets with spontaneous and sporadic viral clearance might be related to specific antibody and T-cell response to HCV proteins in vivo. Replication of CE1E2p7 chimera was observed in primary hepatocyte cultures by immunofluorescent staining in vitro. CONCLUSION: Infectious HCV chimeras causing chronic hepatitis in marmosets might constitute a small primate model suitable for evaluation of virus-cell interaction, vaccination, and antiviral therapy against HCV infection.

Increasing threat of brucellosis to low-risk persons in urban settings, China.

Cases of brucellosis were diagnosed in 3-month-old twins and their mother. An epidemiologic survey suggested that raw sheep or goat meat might be the source of Brucella melitensis infection. This finding implies that the increasing threat of brucellosis might affect low-risk persons in urban settings in China.

Characterization of the novel HLA-B*40:01:02:48 allele by next-generation sequencing.

HLA-B*40:01:02:48 differs from HLA-B*40:01:02:01 by one nucleotide substitution C to A at position -138 in 5'UTR.

Characterization of the novel allele, HLA-B*40:509Q in a Chinese individual.

HLA-B*40:509Q differs from HLA-B*40:01:02:01 by a three nucleotide deletion at position 764 to 766 in exon 4.

Description of two new HLA alleles: HLA-DRB1*14:234 and HLA-DRB1*14:240 in Chinese individuals.

Compared with HLA-DRB1*14:54:01:01, the alleles HLA-DRB1*14:234 and HLA-DRB1*14:240 each show one nucleotide substitution.

The novel HLA-DPA1*02:86 allele was identified by next-generation sequencing.

HLA-DPA1*02:86 differs from HLA-DPA1*02:01:01 by a single nucleotide substitution at position 680 C > A in exon 4.

Identification of the novel HLA-DRB1*11:298 allele in a Chinese cord blood donor.

HLA-DRB1*11:298 shows one single nucleotide substitution at position 397 T>G compared with HLA-DRB1*11:01:01:01.

Identification of the novel HLA-DQB1*06:466 allele by next-generation sequencing in a Chinese cord blood donor.

HLA-DQB1*06:466 differs from HLA-DQB1*06:01:01:01 by a single nucleotide substitution at position 571G→A.

Identification of the novel HLA-A*24:567N allele in a Chinese individual.

HLA-A*24:567N differs from HLA-A*24:02:01:01 by a single nucleotide deletion at position 149.

Characterization of the novel HLA-DPB1*1299:01N allele by next-generation sequencing.

HLA-DPB1*1299:01N differs from HLA-DPB1*427:01 by one single nucleotide substitution and one nucleotide insertion.

Characterization of the novel HLA-C*06:02:96 allele by next-generation sequencing.

HLA-C*06:02:96 differs from HLA-C*06:02:01:01 by one single nucleotide substitution at position 924 C > A.

Description of two new HLA-C alleles, HLA-C*15:245 and HLA-C*15:246, identified in Chinese individuals.

Two novel HLA alleles HLA-C*15:245 and HLA-C*15:246 alleles detected during routine next generation sequencing.

Identification of the novel HLA-B*50:01:17 allele by polymerase chain reaction sequence-based typing.

HLA-B*50:01:17 shows one nucleotide substitution at position 861 when compared with HLA-B*50:01:01:01.

The novel HLA-C allele, HLA-C*07:02:141 was identified in a Chinese individual.

HLA-C*07:02:141 shows one nucleotide substitution at position 4 when compared to HLA-C*07:02:01:01.

Description of two new HLA alleles, HLA-A*26:01:70 and HLA-A*26:01:74 in Chinese individuals.

Compared with HLA-A*26:01:01:01, the alleles HLA-A*26:01:70, and HLA-A*26:01:74 each show one nucleotide substitution, respectively.

Identification of the novel HLA-DPB1*03:01:14 allele by next-generation sequencing in a Chinese cord blood donor.

HLA-DPB1*03:01:14 differs from HLA-DPB1*03:01:01:01 by a single nucleotide substitution at position 390 C > A.

Identification of the novel allele, HLA-DRB1*08:03:12, in a Chinese cord blood donor.

HLA-DRB1*08:03:12 differs from HLA-DRB1*08:03:02 by a single nucleotide substitution at position 702C>T.