Characterization of fertilization-blocking monoclonal antibody 1G12 with human sperm-immobilizing activity.
A mouse hybridoma (1G12) producing sperm-immobilizing MoAb to human sperm was established and characterized in order to study the antigens relevant to sperm immobilization by antibodies. MoAb 1G12 had strong sperm-immobilizing and agglutinating activities and also showed a fertilization-blocking activity on in vitro fertilization tests. The antibody absorption experiments showed that MoAb 1G12 reacted not only to ejaculated sperm but also human seminal plasma, suggesting that the corresponding antigen might be a sperm coating antigen. The MoAb also reacted with peripheral blood lymphocytes. In histochemical studies, the epithelia of corpus epididymis were most strongly stained. Ejaculated sperm were stained with a granular pattern for their entire surface by immunofluorescence. MoAb 1G12 recognized polymorphic glycoproteins of 15-25 kD in the ejaculated sperm extract in Western blot analysis. After deglycosilation of the sperm extract, only a single staining band of under 15 kD was detected by MoAb 1G12. This suggests that the antigen epitope recognized by MoAb 1G12 might be a peptide of the core portion of the glycoprotein. MoAb 1G12 might be a useful tool for studying the mechanism of egg-sperm interaction, and also be applied to identifying the corresponding antigen by using gene technology.
Komori
S
S
Department of Obstetrics and Gynecology, Hyogo College of Medicine, Nishinomiya, Japan.
Kameda
K
K
Sakata
K
K
Hasegawa
A
A
Toji
H
H
Tsuji
Y
Y
Shibahara
H
H
Koyama
K
K
Isojima
S
S
D000042:Absorption / Q000276:immunology; D000372:Agglutination Tests; D000818:Animals; D019138:Antibodies, Blocking / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D004822:Epididymis / Q000276:immunology; D005307:Fertilization in Vitro; D019084:Fluorescent Antibody Technique, Indirect; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D008214:Lymphocytes / Q000276:immunology; D008297:Male; D008698:Mesylates / Q000494:pharmacology; D051379:Mice; D008808:Mice, Inbred CBA; D010504:Periodic Acid / Q000494:pharmacology; D012661:Semen / Q000276:immunology; D013094:Spermatozoa / Q000276:immunology*
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D008698:Mesylates; D010504:Periodic Acid; C009288:metaperiodate; C012077:trifluoromethanesulfonic acid