PMID- 35791899 OWN - NLM STAT- MEDLINE DCOM- 20220908 LR - 20220920 IS - 2576-2095 (Electronic) IS - 2096-5451 (Print) IS - 2576-2095 (Linking) VI - 5 IP - 4 DP - 2022 Dec TI - Development of a humanized HLA-A30 transgenic mouse model. PG - 350-361 LID - 10.1002/ame2.12225 [doi] AB - BACKGROUND: There are remarkable genetic differences between animal major histocompatibility complex (MHC) systems and the human leukocyte antigen (HLA) system. HLA transgenic humanized mouse model systems offer a much better method to study the HLA-A-related principal mechanisms for vaccine development and HLA-A-restricted responses against infection in human. METHODS: A recombinant gene encoding the chimeric HLA-A30 monochain was constructed. This HHD molecule contains the following: alpha1-alpha2 domains of HLA-A30, alpha3 and cytoplasmic domains of H-2D(b) , linked at its N-terminus to the C-terminus of human beta2m by a 15-amino-acid peptide linker. The recombinant gene encoding the chimeric HLA-A30 monochain cassette was introduced into bacterial artificial chromosome (BAC) CH502-67J3 containing the HLA-A01 gene locus by Red-mediated homologous recombination. Modified BAC CH502-67J3 was microinjected into the pronuclei of wild-type mouse oocytes. This humanized mouse model was further used to assess the immune responses against influenza A virus (H1N1) pdm09 clinically isolated from human patients. Immune cell population, cytokine production, and histopathology in the lung were analyzed. RESULTS: We describe a novel human beta2m-HLA-A30 (alpha1alpha2)-H-2D(b) (alpha3 transmembrane cytoplasmic) (HHD) monochain transgenic mouse strain, which contains the intact HLA-A01 gene locus including 49 kb 5'-UTR and 74 kb 3'-UTR of HLA-A01*01. Five transgenic lines integrated into the large genomic region of HLA-A gene locus were obtained, and the robust expression of exogenous transgene was detected in various tissues from A30-18# and A30-19# lines encompassing the intact flanking sequences. Flow cytometry revealed that the introduction of a large genomic region in HLA-A gene locus can influence the immune cell constitution in humanized mice. Pdm09 infection caused a similar immune response among HLA-A30 Tg humanized mice and wild-type mice, and induced the rapid increase of cytokines, including IFN-gamma, TNF-alpha, and IL-6, in both HLA-A30 humanized Tg mice and wild-type mice. The expression of HLA-A30 transgene was dramatically promoted in tissues from A30-9# line at 3 days post-infection (dpi). CONCLUSIONS: We established a promising preclinical research animal model of HLA-A30 Tg humanized mouse, which could accelerate the identification of novel HLA-A30-restricted epitopes and vaccine development, and support the study of HLA-A-restricted responses against infection in humans. CI - (c) 2022 The Authors. Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences. FAU - Zhu, Meng-Min AU - Zhu MM AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Niu, Bo-Wen AU - Niu BW AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Liu, Ling-Ling AU - Liu LL AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Yang, Hua AU - Yang H AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Qin, Bo-Yin AU - Qin BY AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Peng, Xiu-Hua AU - Peng XH AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Chen, Li-Xiang AU - Chen LX AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Liu, Yang AU - Liu Y AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Wang, Chao AU - Wang C AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Ren, Xiao-Nan AU - Ren XN AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Xu, Chun-Hua AU - Xu CH AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Zhou, Xiao-Hui AU - Zhou XH AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. FAU - Li, Feng AU - Li F AUID- ORCID: 0000-0003-1291-5315 AD - Department of Laboratory Animal Science, Shanghai Public Health Clinical Center, Shanghai, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220706 PL - United States TA - Animal Model Exp Med JT - Animal models and experimental medicine JID - 101726292 RN - 0 (HLA-A Antigens) RN - 0 (HLA-A*30 antigen) SB - IM MH - Animals MH - *Disease Models, Animal MH - *HLA-A Antigens MH - Humans MH - Influenza A Virus, H1N1 Subtype MH - Mice MH - *Mice, Transgenic PMC - PMC9434587 OTO - NOTNLM OT - HLA-A30 OT - humanized mouse OT - immunology OT - major histocompatibility complex (MHC) COIS- The authors declare that they have no conflicts of interest relevant to this work. We declare that we do not have any commercial or associative interest that represents a conflict of interest in connection with the work submitted. Xiao-hui Zhou is an Editorial Board member of AMEM and a co-author of this article. To minimize bias, he was excluded from all editorial decision-making related to the acceptance of this article for publication. EDAT- 2022/07/07 06:00 MHDA- 2022/09/09 06:00 PMCR- 2022/07/06 CRDT- 2022/07/06 05:52 PHST- 2022/03/17 00:00 [revised] PHST- 2021/09/03 00:00 [received] PHST- 2022/03/18 00:00 [accepted] PHST- 2022/07/07 06:00 [pubmed] PHST- 2022/09/09 06:00 [medline] PHST- 2022/07/06 05:52 [entrez] PHST- 2022/07/06 00:00 [pmc-release] AID - AME212225 [pii] AID - 10.1002/ame2.12225 [doi] PST - ppublish SO - Animal Model Exp Med. 2022 Dec;5(4):350-361. doi: 10.1002/ame2.12225. Epub 2022 Jul 6.