Malays. Appl. Biol. (June 2003) 32(1): 15-20
THE EFFECT OF Ml MONOCLONAL ANTIBODY ON IN VIVO FERTILIZATION IN HAMSTERS
*MAHANEM, M.N. and DZULSUHAIMI, D.
School of Biosciences & Biotechnology, Faculty of Science & Technology Universiti Kebangsaan Malaysia, 43600 Bangi Selangor, Malaysia
ABSTRACT
The Ml monoclonal antibody (mAb) recognizes an antigenic protein, localized at the equatorial segment of the hamster sperm head. A previous study had shown that Ml mAb blocked sperm-oocyte fusion in hamster. In this experiment, passive immunization of female hamsters were conducted to study the effect of Ml mAb on in vivo fertilization. Superovulated female hamsters received two doses of antibody injections, intraperitoneally (at time similar to the PMSG & hCG injections schedule) at concentrations of 0.5, 1.0, 2.0, 4.0 and 8.0 mg Ml mAb respectively. Two weeks after mating with proven fertile male hamster, the females were killed and implantation sites were counted. Ml mAb at 2.0, 4.0 and 8.0 mg showed a significant decrease in number of implantation sites (P< 0.05) compared to 0.5, 1.0 mg Ml mAb and the control group. Histological studies on ovaries and uterus showed no evidence of pathological changes in control and treated females. These results suggest a possible basis for the development of Ml antigen as a contraceptive vaccine, employing specific sperm antigens that participate in the fertilization process.
ABSTRAK
Antibodi monoklon Ml yang mengenali protein antigenik Ml berada di segmen khatulistiwa kepala sperma hamster. Kajian terdahulu menunjukkan antibodi monoklon Ml telah menghalang pelakuran sperma-oosit hamster. Dalam kajian ini imunisasi pasif ke atas hamster betina telah dilakukan untuk mengkaji kesan antibodi monokoln terhadap persenyawaan in vivo. Hamster betina yang dirangsang ovulasi disuntik dengan dua dos suntikan antibodi secara intraperitoneal (pada masa yang sama dengan suntikan PMSG dan hCG) pada kepekatan 0.5, 1.0, 2.0, 4.0 dan 8.0 mg antibodi monoklon Ml masing-masing. Dua minggu selepas kopulasi dengan hamster jantan yang terbukti subur, haiwan betina dimatikan dan bilangan tapak implantasi ditentukan. Suntikan antibodi monoklon Ml pada 2.0, 4.0 dan 8.0 mg didapati menunjukkan penurunan bilangan tapak implantasi dengan signifikan (P < 0.05) berbanding suntikan antibodi monoklon pada 0.5, 1.0 mg dan kawalan. Kajian histologi ovari dan uterus pula menunjukkan tiada bukti berlaku perubahan patologi dalam kumpulan haiwan betina perlakuan dan kawalan. Keputusan kajian mencadangkan potensi pembangunan antigen Ml sebagai vaksin kontraseptif, menggunakan antigen spesifik sperma yang mengambil bahagian dalam proses persenyawaan.
Key words: Ml monoclonal antibody, passive immunization, in vivo fertilization.
REFERENCES
Allen, C.A., Green, D.P.L. 1995. Monoclonal antibodies which recognizes equatorial segment epitopes presented de novo following the A 23187 induced acrosome reaction of guinea pig sperm. Journal of Cell Science, 108: 767-777.
Bellve, A.R., Moss, R. 1983. Monoclonal antibodies as probes of reproductive mechanism. Biology of Reproduction, 28:1-13.
Cohen, D.J., Ellerman, D.A., Cuasnicu, P.S. 2000. Mammalian sperm-egg fusion, evidence that epididymal protein DE plays a role in mouse gamete fusion. Biology of Reproduction, 63: 462-468.
Diekmann, A.B., Herr, J.C. 1997. Sperm antigens and their use in the development of immunocontraceptive. American Journal Reproductive Immunology, 37: 111-117.
Drury, R.A.B. 1967. Carleton's histological technique. Oxford University Press, London.
Ellis, D.H., Hartman, T.D., Moore, H.D.M. 1985. Maturation and function of the hamster spermatozoon probed with monoclonal antibodies. Journal of Reproductive Immunology, 7: 299-314.
Harlow, E., Lane, D. 1988. Antibodies. A Laboratory Manual Cold Spring Harbor Laboratory Press, Cold Spring Harbor.
Humason, G.L. 1979. Animal tissue techniques. W.H. Freeman and Company, San Francisco.
Kaufman, M.H. 1992. The Atlas of Mouse Development. Academic Press, Cambridge.
Lee, C.Y.G., Wong, E., Zhang, J.H. 1986. Inhibitory effects of monoclonal sperm antibodies on the fertilization of mouse oocytes in-vitro and in-vivo. Journal of Reproductive Immunology, 9: 261-274.
Lee, C.Y.G., Wong, E., The, C.Z., Nishizawa, Y. 1985. Generation of mouse oocyte monoclonal isoantibodies; their effect and those of antisperm monoclonal antibodies on in-vitro fertilization. Journal of Reproductive Immunology, 7: 3-26.
Lowry, O.K., Rosebrough, N.J., Farr, A.I., Randall, R.J. 1951. Protein measurement with the Folin Phenol reagent. Journal of Biological Chemistry, 193: 265-275.
Mat Noor, M., Moore, H.D.M. 1996. Characterization of a putative fusogenic receptor on the equatorial region of hamster spermatozoa. Biology of Reproduction, 54 (1): 98.
Mat Noor, M., Moore, H.D.M. 1999. Monoclonal antibody mat recognizes an epitope of the sperm equatorial region and specifically inhibits sperm-oolema fusion but not binding. Journal of Reproduction and Fertility, 115: 215-224.
Manandhar, G., Toshimori, K. 2001. Exposure of sperm head equatorin after acrosome reaction and its fate after fertilization in mice. Biology of Reproduction, 65: 1425-1436.
Menge, A.C. 1980. Clinical Immunologic infertility. Immunological Aspect oflnfertlity & Fertility Regulation. Elsivier / North Holland. New York.
Moore, H.D.M., Hartman, T.D. 1984. Localization by monoclonal antibodies of various surface antigens of hamster sperm and the effect of antibody on fertilization in vitro. Journal of Reproduction and Fertilility. 70: 175-183.
Naz, R.K., Sacco, A., Singh, O., Pal, R., Talwar, G.P. 1995. Development of contraceptive vaccines for humans using antigens derived from gametes (sperm and zona pellucida) and hormones (human chorionic gonadotrophin): Current status. Human Reproduction Update. 1: 1-18.
Okabe, M., Yagasaki, M., Oda, H., Matzno, S., Kohama, Y., Mimura, T. 1988. Effect of a monoclonal anti-mouse sperm antibody (OBF 13) on the interaction of mouse sperm with
zona-free mouse and hamster eggs. Journal of Reproductive Immunology, 13: 211-219.
Primakoff, P., Hyatt, H., Tredick-Kline, J. 1987. Identification and purification of a sperm surface protein with a potential role in sperm-egg membrane fusion. Journal of Cell. Biology, 104: 141-149.
Rugh, R. 1971. A guide to vertebrate development. 6th Edition. Burgess Publishing Company. Minneapolis.
Saling, P.M., Irons, G., Waibel, R. 1985. Mouse sperm antigens that participate in fertilization (I) Inhibition of sperm fusion with the egg plasma membrane using monoclonal antibodies. Biology of Reproduction, 33: 515-526.
Saling, P.M., Waibel, R. 1985. Mouse sperm antigens that participate in fertilization (II) Passive immunization with a single monoclonal antisperm antibody inhibits pregnancy and fertilization in-vivo. Biology of Reproduction, 33: 537-544.
Toshimori, K., Tanii, I., Araki, S., Oura, C. 1992. Characterization of the antigen recognized by a monoclonal antibody MN9: unique transport pathway to the equatorial segment head during spermiogenesis. Cell and Tissue Research, 270: 459-468.
Tung, K.S.K., Goldberg, E.H., Goldberg, E. 1979. Immunobiological consequences of immunization of female mice with homologous spermatozoa; induction of fertility. Journal of Reproductive Immunology, 1: 145-158.
Yoshinaga, K., Saxena, D.K., Oh-oka, T., Tanii, T., Toshimori, K. 2001. Inhibition of mouse fertilization by intra- oviductal injection of anti-equatorin monoclonal antibody. Reproduction, 122: 649-655.





