[Mouse anti-human CXCR3 mAb preparation and its biological function]

Obtain hybridoma cell line with continuing expression of mouse anti-human CXCR3 mAb, investigate expression characteristics of human CXCR3 and how CXCR3 signal transduction function on L929-huCXCR3 and colon carcinoma cell lines transfer and growth.

Taking L929-huCXCR3 cell with high expression of human CXCR3 membrane molecule as immunogen to immunize BALB/c mouse, we fused immunized mouse spleen cell with myeloma cell Sp2/0 of its same germ line, then used L929-huCXCR3 as screening cell and empty vector transfected cell L929-mock as negative control. Obtained hybridoma cell line with continuing secretion of anti-human CXCR3 mAb through flow cytometry. We used Ig subclass type rapid qualitation indicator paper method and indirect immunofluorescence to identify obtained hybridoma cell line and mAb, indirect immunofluorescence to analyze CXCR3 expression on tumor cell surface, Transwell isolation cabin to assess effect on L929-huCXCR3 and colon carcinoma cell line Colo205, HCT116 and HT29 migration by mAb, MTT method to analyze how mAb function on colon carcinoma cell line Colo205 proliferation.

Obtained a hybridoma cell line with continuing secretion of mouse anti-human CXCR3 mAb, named 9B5. According to rapid qualitation test paper analysis, light chain of the mAb was chain and heavy chain is IgG1 subclass. Indirect immunofluorescence and flow cytometry results show that the mAb can recognize CXCR3 molecules on the surface of activated T lymphocyte and colon carcinoma cell line Colo205, HCT116 and HT29 cell. mAb 9B5 can inhibit oriented migration of L929-huCXCR3 cells, colon carcinoma cell line Colo205, HCT116 and HT29 cell, it can also inhibit Colo205 growth promotion effect by IP-10.

Successfully obtain a hybridoma cell line with continuing secretion of mouse anti-human CXCR3 mAb, which has laid material foundation on investigation of CXCR3 expression characteristics and CXCR3 signal transduction function on tumor growth and migration. It is prospective to create a new way and a new drug for treatment of tumor metastasis.