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Human Reproduction, Vol. 15, No. 6, 1329-1337, June 2000
© 2000 European Society of Human Reproduction and Embryology

Leukocyte detection in human semen using flow cytometry

Giuseppe Ricci1,3, Gianni Presani2, Secondo Guaschino1, Roberto Simeone1 and Sandra Perticarari2

1 Department of Obstetrics and Gynaecology, University of Trieste, Istituto per l'Infanzia `Burlo Garofolo', IRCCS and 2 Clinical Pathology Laboratory, Istituto per l'Infanzia `Burlo Garofolo', IRCCS, 34137 Trieste, Italy

This study set out to establish a new method, using flow cytometry, to evaluate leukocytes in semen. Ejaculates of 59 males, asymptomatic for genitourinary infections, were examined. Routine semen analyses were carried out as well as peroxidase and polymorphonuclear granulocyte–elastase detection. Leukocytes were detected combining flow cytometry and monoclonal antibodies (anti-CD45, anti-CD53). This technique reliably assessed the total number of leukocytes and differentiated subpopulations even at low concentrations. The peroxidase test and elastase determination showed good specificity, but only moderate sensitivity versus flow cytometry combined with monoclonal antibodies. No significant association was observed between semen parameters and leukocytospermia whether evaluated by conventional methods or flow cytometry except for a moderate correlation between spermatozoa and CD53-positive cell concentrations. A first comparison of data from patients grouped on the basis of leukocytospermia (>106 white blood cells, WBC/ml) or non-leukocytospermia revealed no significant differences in semen parameters; lowering the threshold value for leukocytospermia to 2x105 WBC/ml, sperm concentration was reduced in the group with a low number of WBC identified by monoclonal antibodies. Flow cytometry using monoclonal antibodies was seen to be a simple, reproducible method that enables leukocytes in semen to be accurately detected and to identify WBC subpopulations without preliminary purification procedures.

Key words: elastase/flow cytometry/human semen/leukocyte/peroxidase

3 To whom correspondence should be addressed. E-mail: ricci{at}burlo.trieste.it


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