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Peripheral blood lymphocytes isolated from an infertile woman possessing strong sperm immobilizing and agglutinating antibodies were stimulated by culturing with poke-weed mitogen (PWM) and spermatozoa from a healthy donor for 5 days. The stimulated lymphocytes were fused with mouse myeloma NS-1 by PEG-1000. Fused growing hybrid cells were observed in 58 of 96 wells, and 22 of these showed the production of human immunoglobulin. Among the 22, one hybridoma clone (H6-3C4) was found to produce human IgM (lambda) with strong sperm immobilizing and agglutinating activities. The supernatant from the culture medium contained approximately 1.5 microgram IgM/ml and the antibody titers were 5000 SI50 units on sperm immobilization and 1:1600 dilutions on sperm agglutination. The hybridoma H6-3C4 has continuously produced high titers of antibody exhibiting sperm immobilizing and agglutinating activities over 8 months and contains chromosomes of acrocentric type from mouse and metacentric type from human. The monoclonal antibody (Mab) H6-3C4 reacted specifically to human seminal plasma, ejaculated spermatozoa and male accessory gland but not to testis, any other somatic tissues, or secreted fluids tested. Immunofluorescence staining indicated that the antigen corresponding to Mab H6-3C4 was present over the surface of ejaculated spermatozoa. The binding of Mab H6-3C4 to human spermatozoa was blocked by the serum of the patient from whom the lymphocytes were obtained for cell fusion.
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
July 1980,
Nihon Sanka Fujinka Gakkai zasshi,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
December 1980,
Clinical and experimental immunology,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
October 1985,
Nihon Sanka Fujinka Gakkai zasshi,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
February 1987,
Biochemical and biophysical research communications,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
January 1985,
Ji sheng chong xue yu ji sheng chong bing za zhi = Journal of parasitology & parasitic diseases,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
January 1989,
Reproduction, fertility, and development,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
November 1978,
Cellular immunology,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
January 1986,
International journal of fertility,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
June 2003,
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology,
S Isojima, and
K Kameda, and
Y Tsuji, and
M Shigeta, and
Y Ikeda, and
K Koyama
March 1986,
Immunology,
