Construction method of humanized transgenic mouse model of chimeric human HLA-DP genome region

Abstract

The invention belongs to the technical field of biology, and relates to a construction method of a transgenic animal model for expressing a human HLA-DP genome region. The BAC clones an HLADP genome region (DOA-DPA1*0103-DPB1*0401) containing 124kb, BAC-DNA is introduced into fertilized eggs of mice through a prokaryotic microinjection technology after DNA sequencing, pulsed field gel electrophoresis identification and purification, and then the fertilized eggs are transplanted into fallopian tubes of false pregnant mice to obtain positive transgenic animals. Hybridizing with wild type mice isconducted to obtain filial generations, and identifying and analyzing are conducted through molecular biology, immunology and histology technologies to finally obtain five transgenic mouse lines integrating different human HLADP genome regions. The mouse model constructed by the method has high-level expression of HLA-DPA1 and HLA-DPB1 on main tissues and organs such as blood cells, spleen and lung, and can be used for T cell epitope and immune pathopoiesis and immune protection effect research of epitope-specific T cells.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for constructing a humanized transgenic mouse model of a chimeric human HLA-DP genome region, in particular to a human MHC (HLA-DPA1*0103-DPB1*0401) transgenic mouse model. build method. Background technique [0002] The prior art discloses that the major histocompatibility complex (MHC) is a group of genes that determine whether the transplanted tissue is compatible, is closely related to the immune response, and is closely linked. In humans, MHC is called human leukocyte antigen (HLA) gene complex. Studies have disclosed that the various genes that make up MHC are distributed in clusters on the short arm of human chromosome 6, mainly including HLA class I, HLA class II and HLA class III genes, of which HLA class II genes mainly include HLA-DP HLA-DQ, HLA-DR and other three types of gene clusters, which encode HLA class II molecules HLA-DP, HLA-DQ, HLA-DR, etc.; HLA class I...

AI Technical Summary

Problems solved by technology

Compared with the DQ and DR loci, there are fewer humanized transgenic mouse models for the DP locus, and only three DP transgenic mice (HLA-DPB1*0401, HLA-DPB1*0201, HLA-DPB1 *1701), for the study of the molecular mechanism of human chronic beryllium disease (Tarantino-Hutchison et al. 2009, Mack et al. 2014); there are many genetic linkage disequilibrium phenomena in the HLA-DP gene; February 2013 WHO IMGT / HLA database (http : / / www.ebi.ac.uk / imgt / hla / , Release3.11.0) published HLA-DPA1 and HLA-DPB1 alleles were 36 and 159; however, HLA-DPA1*0103 / DPB1* 0401 (DP401) and HLA-DPA1*0103 / DPB1*0402 (DP402) two genotypes are distributed in 20-60% of the world population, and DP401 and DP402 share similar antigenic determinants (Castelli et al. 2002, JUNBAO et al. 2005 , Sidney et al. 2010); and studies have shown that HLA-DP4 can present various polypeptides from influenza virus, HIV virus, and tumor cells to CD4 cells (Castelli et al. 2002, JUNBAO et al. 2005, Cohen et al. 2006, Sidney et al. 2010)

Images