ABSTRACT
Generation of free radicals in ejaculate samples from infertile patients was evaluated using chemiluminescent technique. The presence of antisperm antibodies in samples increased the possibility of damages to spermatozoon plasma membranes due to excessive generation of free radicals.
Subject(s)
Free Radicals/metabolism , Infertility, Male/metabolism , Spermatozoa/metabolism , Antibodies/immunology , Cell Survival , Humans , Infertility, Male/immunology , Male , Spermatozoa/immunologyABSTRACT
Conflicting opinions about the effects of antisperm antibodies on fertilization can be due to inadequacy of experimental approaches in evaluating the antisperm immunity. Detection of antisperm antibodies bound to the surface of live spermatozoa can be associated with aggregation of surface antigen-antibody complexes followed by metabolic activation of spermatozoa and acrosomal reaction, which impair cell resistance. New concept of antisperm immunity and its influence on reproduction can be formulated only after comprehensive studies of the mechanisms of spermatozoon response to binding of antisperm antibodies. Further improvement of quantitative assays of antisperm antibodies and evaluation of their effect on spermatozoon function should be aimed at selection of experimental conditions preventing changes in spermatozoa coated with antisperm antibodies during in vitro manipulations.
Subject(s)
Antibodies/immunology , Fertilization/physiology , Infertility/immunology , Spermatozoa/immunology , Acrosome Reaction/physiology , Antibodies/analysis , Antigen-Antibody Complex/metabolism , Female , Flow Cytometry/methods , Humans , Immunologic Tests , Male , Spermatozoa/metabolismABSTRACT
Flow cytometry (FCM) analysis of live antibody-coated spermatozoa subjected to immunofluorescence staining (FCM test) is considered an objective method for the quantitative detection of antisperm antibodies (ASA). But the cross-linking of cell surface antigen (Ag) with bivalent antibodies and/or antigen-antibody (Ag-Ab) complexes with second antibodies may induce the reorganization of surface components (patching and capping) and result in their shedding from the sperm surface. The present study estimates the relationship between aggregation of Ag-Ab complexes on the sperm surface and the results of indirect FCM test. Swim-up spermatozoa of normozoospermic men were incubated with ASA-positive sera from infertile patients and with second antibodies fluorescein isothiocyanate (FITC)-labelled goat anti-human IgG polyclonal antiserum under different conditions and then analysed by FCM and fluorescence microscopy. It was shown that low temperature, cytochalasin B, excess or lack of the primary and/or secondary antibodies and sperm fixation by paraformaldehyde may inhibit aggregation and shedding of Ag-Ab complexes and dramatically increase ASA quantity determined on the sperm surface. However, inhibition of aggregation on the live sperm surface was observed only in a minority of ASA-positive samples and was poorly reproducible using semen of different donors. A high probability of Ag-Ab complex shedding from the sperm surface during experimental manipulation limits the use of indirect FCM test for quantitative ASA determination.
Subject(s)
Antigen-Antibody Complex/analysis , Autoantibodies/blood , Flow Cytometry , Spermatozoa/immunology , Cytochalasin B/pharmacology , Fluorescein-5-isothiocyanate , Fluorescent Antibody Technique, Indirect , Fluorescent Dyes , Humans , Immunoglobulin G/immunology , Infertility, Male/immunology , Male , Microscopy, Fluorescence , Reproducibility of Results , Sperm CountSubject(s)
Antibodies/blood , Infertility, Female/immunology , Infertility, Male/immunology , Spermatozoa/immunology , False Negative Reactions , Female , Humans , Infertility, Female/diagnosis , Infertility, Male/diagnosis , Insemination, Artificial, Heterologous , Insemination, Artificial, Homologous , Latex Fixation Tests , MaleABSTRACT
OBJECTIVE: To investigate the influence of sera and peritoneal fluids (PFs) from fertile and infertile women on the binding of antisperm antibodies to the surface of spermatozoa. DESIGN: The immunoglobulin (Ig) G antisperm antibodies binding to the surface of liver spermatozoa was evaluated after their incubation in antisperm antibodies-positive serum from an infertile male in the presence and absence of female sera or PFs. SETTING: Russian Scientific Center for Obstetrics, Gynaecology, and Perinatology. PATIENT(S): Serum and PF samples from fertile and infertile women; antisperm antibodies-positive serum from infertile men; high-quality fresh semen from healthy donors. INTERVENTION(S): Serum samples were obtained from fertile and infertile women and from infertile men. Peritoneal fluids were collected during routine laparoscopy. MAIN OUTCOME MEASURE(S): The proportion of spermatozoa positive for IgG antibodies and the quantity of antisperm antibodies on the sperm surface measured by flow cytometry (FCM). RESULT(S): The addition of sera from fertile or infertile women with endometriosis or pelvic adhesion disease to an IgG antisperm antibodies-positive male serum resulted in significant inhibition of the antisperm antibodies binding to the sperm surface. CONCLUSION(S): Sera of fertile as well as infertile woman contain factors that block IgG antisperm antibodies binding to the surface of live spermatozoa.
