An isoimmune response to human sperm clathrin in an infertile woman with systemic lupus erythematosus.

We have employed a proteomic approach to study the immune response to human sperm in an infertile female patient suffering from systemic lupus erythematosus (SLE). Human sperm antigenic extracts were resolved by means of two-dimensional electrophoresis and electroblotted onto nitrocellulose membranes. The membranes were incubated with serum from the SLE patient. Sperm antigens that were reactive to polyclonal antibodies were next visualized on X-ray film, using the enhanced chemiluminescence (ECL). Three spots corresponding to the positions of sperm immunoreactive antigens on a nitrocellulose membrane were localized in a silver stained gel and subjected to mass spectrometry. A database search of the sequences recognized by the analyzed SLE serum revealed its homology to the clathrin heavy chain (CHC). Further analysis revealed that anti-CHC antibody reacted with multiple sperm antigenic determinants, resolved by either one- or two-dimensional electrophoresis. When studied by immunofluorescence, we demonstrated anti-CHC antibody reactivity with the sperm tail tip (corresponding to the sperm agglutination pattern), also with the principal piece and with cytoplasmic droplets around the sperm midpiece. Live sperm clearly exhibited reactivity with the midpiece. This study demonstrates clathrin heavy chain on human sperm using serum of an infertile individual with a concomitant autoimmune disease.