The Immune Effect In Vitro Of Hspgp96-peptide Complex Derived From Human Renal Tumor Tissues

Objective: Carcinoma of kidney was a kind of the common tumor in urinary system, it has strong cancerometastasis and immunity. The therapy of the metastasis carcinoma of kidney was to become one of the clinical problems. As recent investigation shown, Heat shock protein (HSP) derived from tumor tissue, especially HSPgp96-peptide complex (HSPPC-96), can elicit antigen-specific cytolytic T-lymphocytes against these tumors. The HSPPC-96 vaccine is in a pivotal phaseIII clinical trial for renal cancer at 80 clinical sites worldwide and have gained good effect. Since the concrete component and activity of the pure polypeptide which have genuine action in HSPPC-96 vaccine is not certainly, Try to find out its activity mechanism we derive HSPgp96 from human carcinoma of kidney, and detect the immunologic reaction of INF-r stimulated by HSPPC-96 and pure polypeptide in vitro, which will help to further comprehend the antigen peptide structure and man-made immunotherapy.Methods: 12 renal tissues of the carcinoma of kidney patients who were accepted radical nephrectomy, renal tissues form 5 healthy individuals, who were accepted kidney Resection because of trauma in our hospital form 2001 to 2004 were collected. we used three-step purification to derive HSPgp96 from human carcinoma of kidney, and use Elispot to detect the immunologic reaction of INF-r stimulated by HSPPC-96 and pure polypeptide in vitro.Results: (1) Western blotting was used to identify the purified proteins, and it confirmed that the proteins are HSPgp96. (2) The HSPgp96-peptide complex and polypeptide derived the renal carcinoma tissues can produce the significantly more frequence of IFN-γ ~+T cell than normal renal tissues(p<0.01). (3) The HSPgp96-peptide complex derived from the renal carcinoma tissues can produce the maximum frequence of IFN- γ ~+T cell, and the frequence was significantly more than stimulus.Conclusion: :Using three-step purification can obtain HSPgp96-peptide complex of high purity;The HSPgp96-peptide complex derived from human renal carcinoma tissues have the significantly strongest immune effect in vitro.