The objectives were to standardize some methodological and analytical aspects of a direct technique to detect sperm-bound anti-sperm antibodies ASAs in bovine semen using flow cytometry, including the effects of pre-fixation of sperm membranes with formalin buffer solution and inclusion of dead cells in the analysis. Fixation of sperm membranes had no significant effect on the percentage of IgG- or IgA-bound spermatozoa detected by flow cytometry. However, including dead cells in the analysis increased the percentage of IgG-bound spermatozoa in fixed live and dead Intra-sample, intra-assay and inter-assay coefficients of variation CV were 0. Flow cytometry correctly identified ASA-positive bulls. Confocal laser microscopy confirmed binding of ASAs to sperm heads and cytoplasmic droplets, and less frequently to midpieces and principal piece.
Antigenic Determinants on the Sperm Cell Surface | SpringerLink
The overall objective of this series of experiments is to generate immunological markers that may elucidate bull sperm surface changes in vitro. Here we report the initial experiments of the study, involving the production and characterization of monoclonal antibodies mAbs again bull sperm. The mAbs were specific to antigens of either the posterior tail or the head regions of bull sperm and detected five major domains of antigen localization in the bull sperm apical crescent, equatorial band, principal acrosomal, whole head, and posterior tail. Eleven of the 13 head-specific mAbs recognized intra-acrosomal antigens, whereas 2 mAbs recognized antigens that were localized in the plasma membrane. They were all sperm specific, with no cross-reactivity to bovine oocytes or to any of the 12 bovine somatic tissues tested. The mAbs were not species specific, however, because 11, 10, 2, and 1 of the 15 mAbs cross-reacted with sheep, pig, mouse, and human sperm, respectively.
Standardization of a method to detect bovine sperm-bound anti-sperm antibodies by flow cytometry
Kim, J. Parrish, H. Momont, D. To test the hypothesis that bull antisperm antibodies have the capacity to interfere with fertilization, antisperm antibodies were generated in three mo-old Holstein bulls by auto-immunizing each bull with sperm three times. All bulls produced serum antisperm IgG1 and IgG2 antibodies.
The Sperm Cell pp Cite as. The presence of HLA antigens on human sperm has been demonstrated by cytotoxic assay, immunofluorescence and inhibition-of-motility assay Fellous et al. Similar results were found for SLA antigen on boar sperm Vaiman ,