Experimental study on the antitumor effect of mouse B7-1 gene

Abstract

Mouse B7-1 cDNA was cloned by RT-PCR from BALB/C mouse splenic cells and inserted into pcDNA3 to construct an eukaryotic expression vector. This constructor was named pCD-mB7-1. in which the B7-1 cDNA was identified to be consistent with the data from other researchers. pCD-mB7-1 plasmid was transfected into B16(F0) cells, and effective expression of mB7-1 in these tumor cells could be detected till the 6th month by RT-PCR and RNA hybridization. Specific cytotoxity assay of lymphocytes was conducted after culturing with tumor cells and the results demonstrated that B16 cells transfected with B7-1 gene were more effective than B16-wt and B16-neo in inducing specific cytotoxity of lymphocytes against B16-wt cells. It is suggested that expression of B7-1 gene in tumor cells could enhance the immunogenicity and induce the effective antitumor immunity.

Keywords

CD80(B7-1) / gene expression / RT-PCR / tumor gene therapy

References

[1] TravisJ. A stimulating new approach to cancer treatment. Science, 1993, 259: 310-310

[2] ChenL, AsheS, BradyW A, et al. . Costimulation of antitumor immunity by the B7 counter-receptor for the T lymphocyte molecules CD28 and CTLA-4. Cell, 1992, 71: 1093-1093

[3] FreemanG J, GrayG S, GimmiC D, et al. . Structure, expression, and T cell costimulatory activity of the murine homologue of the human B lymphocyte activation antigen B7. J Exp Med, 1991, 174: 625-625

[4] TokunageK, TaniguchiH, YodaK, et al. . Nucleotide sequence of a full-length cDNA for mouse cytoskeletal beta-actin mRNA. Nucleic Aicds Res, 1986, 14(6): 2829-2829

[5] ChomciynsskiP, PiotrC, NicolettaSacchi. Single step method of RNA isolation by acid guanidinum thiocyanate phenol-chloro-form extration. Anal Biochem, 1987, 162: 156-156

[6] 1992.

[7] LDH 1989, 9(4):218

[8] InobeM, LinsleyP S, LedbetterJ A, et al. . Identification of an alternatively spliced form of the murine homologue of B7. Biochem Biophys Res Commun, 1994, 200(1): 443-443

[9] SchwartzR H. A cell culture model for T lymphocyte clonal anergy. Science, 1990, 248: 139-139

[10] GimmiC D, FreemanG J, GribbenJ G, et al. . Human T-cell clonal anergy is induced by antigen presentation in the absence of B7 costimulation. Proc Natl Sci USA, 1993, 90: 6586-6586

[11] van der BruggenP, TraversariC, ChomezP, et al. . A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science, 1991, 254: 1643-1643

[12] LiY, McGowanP, HellstromI, et al. . Costimulation of tumor-reactive CD4⁺ and CD8⁺ T lymphocytes by B7, a natural ligand for CD28, can be used to treat established mouse melanoma. J Immunol, 1994, 153(1): 421-421