Research for T epitope of a zona pellucida and application for vaccine

1997 Fiscal Year Final Research Report Summary

• Project/Area Number: 08660353
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied animal science
• Research Institution: GRANT-IN-AID FOR SCIENTIFIC RESEARCH (C) (2)
• Principal Investigator: SHIGETA Minoru INSTITUTION ; HYOGO COLLEGE OF MEDICINE. DEPARTMENT ; MEDICINE . TITLE OF POSITION ; ASSOCIATED PROFESSOR, 医学部, 助教授 (80122315)
• Co-Investigator(Kenkyū-buntansha): HASEGAWA Akiko INSTITUTION ; HYOGO COLLEGE OF MEDICINE. DEPARTMENT ; MEDICINE . TITLE OF POSITI, 医学部, 助手 (50212402)
• Project Period (FY): 1996 – 1997
• Keywords: porcine zona pellucida / T cell epitope / vaccine

Research Abstract

The aima of the present study is to establish the porcine zona pellucida specific antigen reactive T cells for the analysis of T cell epitope correlated with the specific B cell epitope as a potential candidate for a contraceptive vaccine. Recombinant porcine zona pellucida protein (r-pZP1) was purified from recombinant fusion proteins for porcine zona pellucida 1 (1-198) . The rec-pZP1 reactive T cell clones (r-pZP1T-2) was obtained with slight modifications of previously described technology (Shigeta, Fathman, 1981) . The r-pZP1T-2 was antigen specific and genetically restricted to H-2^d phenotype. The r-pZP1T-2 was proliferated with r-pZP1 (198mer) and synthetic peptides-TLK (20mer) but not with CTY (18mer) nor KRV (31mer). These results show that there is at least one T cell epitope within TLK peptide (20mer) but there is no T cell epitope within CTY or KRV for the r-pZP1T-2. Peptides were synthesized in a manner to remove amino acids one by one from the 20-amino acid peptide (TLK) . Proliferative response of the r-pZP1T-2 with these peptides suggested that the 7mer motif of 65-71 YEACTKR seems to be essential for the epitope of the rec-pZP1T-2. B cell epitope for MAb-5H4 which inhibits the pig sperm-ZP interaction was determined to be present on the amino acid No.54-61 of rec-pzp1. Therefore, it is easy to speculate that 7mer motif of 65-71 for T cell epitope is closely related to 8mer motif of 54-61 for B cell epitope and help to induce helper T cells for antibody production by B cell for MAb-5H4 epitope.