Background : There are remarkable genetic differences between animal major histo-compatibility complex (MHC) systems and the human leukocyte antigen (HLA) sys-tem. 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 con-structed. 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 chro-mosome (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 his-topathology 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 in-fection 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-i nfection (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.

Meng-min Zhu;Bo-wen Niu;Ling-ling Liu;Hua Yang;Bo-yin Qin;Xiu-hua Peng;Li-xiang Chen;Yang Liu;Chao Wang;Xiao-nan Ren;Chun-hua Xu;Xiao-hui Zhou;Feng Li

Department of Laboratory Animal Science,Shanghai Public Health Clinical Center,Shanghai,China

动物模型与实验医学（英文）

[1]Meng-min Zhu;Bo-wen Niu;Ling-ling Liu;Hua Yang;Bo-yin Qin;Xiu-hua Peng;Li-xiang Chen;Yang Liu;Chao Wang;Xiao-nan Ren;Chun-hua Xu;Xiao-hui Zhou;Feng Li-.Development of a humanized HLA- A30 transgenic mouse model)[J].动物模型与实验医学（英文）,2022(04):350-361

monochain,CH502,67J3,microinjected,topathology,Pdm09,nfection

Development,humanized,HLA,A30,transgenic,mouse,model,Background,There,are,remarkable,genetic,differences,between,animal,major,histo,compatibility,complex,MHC,systems,leukocyte,antigen,offer,much,better,method,study,related,principal,mechanisms,vaccine,development,restricted,responses,against,infection,Methods,recombinant,encoding,chimeric,was,structed,This,HHD,molecule,contains,following,domains,cytoplasmic,2D,linked,its,terminus,2m,by,amino,acid,peptide,linker,cassette,introduced,into,bacterial,artificial,chro,mosome,BAC,containing,A01,locus,Red,mediated,homologous,recombination,Modified,pronuclei,wild,type,oocytes,further,assess,immune,influenza,virus,H1N1,pdm09,clinically,isolated,from,patients,Immune,cell,population,production,lung,were,analyzed,Results,We,describe,novel,transmembrane,strain,which,intact,including,kb,UTR,Five,lines,integrated,large,genomic,region,obtained,robust,expression,exogenous,transgene,detected,various,tissues,18#,19#,encompassing,flanking,sequences,Flow,cytometry,revealed,that,introduction,can,influence,constitution,mice,caused,similar,among,Tg,induced,rapid,increase,cytokines,IFN,both,dramatically,promoted,days,post,dpi,Conclusions,established,promising,preclinical,research,could,accelerate,identification,epitopes,support,humans

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