Dataset Information

Development of a humanized HLA-A30 transgenic mouse model.

ABSTRACT:

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: α1-α2 domains of HLA-A30, α3 and cytoplasmic domains of H-2D^b , linked at its N-terminus to the C-terminus of human β2m 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 β2m-HLA-A30 (α1α2)-H-2D^b (α3 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-γ, TNF-α, 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.

SUBMITTER: Zhu MM

PROVIDER: S-EPMC9434587 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Publications

Development of a humanized HLA-A30 transgenic mouse model.

Zhu Meng-Min MM Niu Bo-Wen BW Liu Ling-Ling LL Yang Hua H Qin Bo-Yin BY Peng Xiu-Hua XH Chen Li-Xiang LX Liu Yang Y Wang Chao C Ren Xiao-Nan XN Xu Chun-Hua CH Zhou Xiao-Hui XH Li Feng F

Animal models and experimental medicine 20220706 4

<h4>Background</h4>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.<h4>Methods</h4>A recombinant gene encoding the chimeric HLA-A30 monochain was constructed. This HHD molecule contains the followi ...[more]

PMID: 35791899

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Dataset Information

Development of a humanized HLA-A30 transgenic mouse model.

Background

Methods

Results

Conclusions

Publications

Development of a humanized HLA-A30 transgenic mouse model.

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