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Abstract
PH-20 (Spam-1) antigen appears to be a bi-functional sperm plasma membrane protein with hyaluronidase activity and its role is related to cumulus penetration as well as zona binding. (LDH-C4) is a key enzyme, distributed specifically in testis and is highly immunogenic, function of which relates to energy metabolism and capacitation of the sperm. Therefore, in this study, we observed the effect of purified native PH-20 and LDH-C4 antigens on sperm function and morphology in dog. Native PH-20 (46, 32 kDa) and LDH-C4 (36, 30, 28) sub units, purified from dog sperm SDS- extracts were used to test as immunocontraceptive candidates in stray dog. Dogs 1, 2, 3, 4, 5/6 and 7/8 were immunized with 46, 32, 36, 30/28, 46/32, 36/30/28 kDa sub units respectively by i/m route and semen was analysed at weekly intervals. High titre of 3200 - 6400 was achieved in all dogs after three boosters. Immune response in treated dogs was confirmed by increase in Ig level in blood serum and reaction of partially purified serum Ig of immunized dogs with respective antigens in double immune diffusion and immunoblots. Immunization of dogs either with individual PH-20/ LDHC sub units or pooled PH-20/LDHC4 sub units showed a significant effect on sperm count, percent motility, viability, HOST and sperm morphology. All the sperm parameters declined to a significant level (p 4 (36/30/28 kDa) sub units had an immunocontraceptive potential in dog.
Highlights
Immunocontraception is a process of using a reproductive protein to produce a humoral immune response that leads to the infertility in animals for a defined time period
60 45 reaction between partially purified Ig of immunized dogs 1, 2, 3, 4, 5, 6, 7 and 8 against Lactatae dehydrogenase (LDHC4)/PH-20 sub units and respective antigens on 60 day confirmed the production of antibodies (Figures 3 and 4)
A single precipitation line in Double immune diffusion (DID) reaction between partially purified Ig of immunized dogs against PH-20 and LDHC4 sub units and respective antigens indicated the production of antibodies
Summary
Immunocontraception is a process of using a reproductive protein to produce a humoral immune response that leads to the infertility in animals for a defined time period. A few of the sperm specific antigens have been isolated and characterized by biochemical and immunological techniques and genes encoding for some of these antigens have been characterized, cloned and sequenced. Notable among these are FA-1 [2], PH-20 [3,4], SP-10 [5], FA-2 [6], SP-17 [7], NZ-1 [8] and LDH-C4 [4,9,10]. Sperm surface PH-20 antigen has been cloned and studied in a number of species including the guinea pig [13], rat [14], mouse [13], horse [15], macaque and human
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