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Human Reproduction, Vol. 15, No. 12, 2545-2553, December 2000
© 2000 European Society of Human Reproduction and Embryology

Antisperm antibodies detection by flow cytometry is affected by aggregation of antigen–antibody complexes on the surface of spermatozoa*

Marina A. Nikolaeva1, Vladimir I. Kulakov, Irina V. Korotkova, Elena L. Golubeva, Diana V. Kuyavskaya and Gennady T. Sukhikh

Laboratory of Clinical Immunology, Russian Scientific Centre for Obstetrics, Gynecology and Perinatology, Moscow, Russia

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.

Key words: antigen antibody complex/antisperm antibodies/flow cytometry/human spermatozoa

1 To whom correspondence should be addressed at: Laboratory of Clinical Immunology, Russian Scientific Centre for Obstetrics, Gynecology and Perinatology, 117815, Oparin 4, Moscow, Russia. E-mail: anikom{at}online.ru

* Presented at the International Symposium on Male Infertility and Assisted Reproduction, Genk, Belgium, April 22–25, 1998, and at the European Meeting of Immunology and Reproduction, Rome, Italy, October, 28–29, 1999.


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[Abstract] [Full Text] [PDF]


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