2003 Fiscal Year Final Research Report Summary
Development of an immunocontraceptive DNA vaccine using sperm
| Project/Area Number |
12557137
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | THE UNIVERSITY OF TOKUSHIMA |
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Principal Investigator |
MAEGAWA Masahiko The University of Tokushima, University Hospital, Assistant Professor, 医学部・歯学部附属病院, 講師 (80253201)
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| Co-Investigator(Kenkyū-buntansha) |
IRAHARA Minoru The University of Tokushima, School of Medicine, Professor, 医学部, 教授 (20160070)
KIDO Hiroshi The University of Tokushima, Division of Enzyme Institute for Enzyme Research, Professor, 分子酵素学研究センター, 教授 (50144978)
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| Project Period (FY) |
2000 – 2003
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| Keywords | immunocontraceptive vaccine / DNA vaccine / sperm antigen / peptide |
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| Research Abstract |
We previously reported that active immunization of female rats with rSMP-230, residue 1-28 of rSMP-B protein, and 80 kDa human sperm antigen (80 kDa HSA), prevented conception. The purpose of this project was to develop an immunocontraceptive vaccine using an expression vector encoding rSMP-B or 80 kDa HSA. Results are as follows : (l) Compared with the nucleic acid sequence of RSD-1 encoding rSMP-B reported in GenBank"808109" (position ?70-773 : ATAT), 4 nucleotides were different (position 770-777 : CACTCTG), resulting in a difference in three amino acids (His-Met to Pro-Leu-Try). (2) When the p3xFLAG-CMV-14 vector containing RSD-1 was transfected into HEK293 cells, cell extracts of HEK293 cells showed a 20-kD band by Western blotting using an anti-FLAG antibody, but no immunoreaction was found in the lane of culture medium. (3) Active immunization of rabbits with synthetic peptides of 80 kDa HSA elicited significant antibody titers. These antibodies specifically recognized the naive protein in an ELISA and induced in-vitro agglutination of human, rat and monkey sperm. Western blot analysis showed that these antibodies bind to the 80 kDa band of sperm extract
These findings demonstrated that the sperm protein rSMP-B and 80 kDa HSA were pomising candidates for the development of an immunocontraceptive vaccine
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