Development of a technique for monitoring the contamination of human semen samples with leukocytes.
OBJECTIVE
To determine whether receptors for the N-formyl chemotactic peptide, FMLP, exist on the surface of human spermatozoa and regulate reactive oxygen species generation by these cells.


METHODS
Chemiluminescent analysis of reactive oxygen species generation by suspensions of human spermatozoa before and after removal of leukocytes using a magnetic cell separation technique.


METHODS
Academic Research Institute.


METHODS
Unselected male volunteers.


RESULTS
Human sperm suspensions responded to FMLP and phorbol ester (PMA) with a burst of reactive oxygen species production. Autoradiographic analyses employing 3H FMLP and chemiluminescence studies involving the selective removal of leucocytes with anti-CD 45-coated magnetic beads demonstrated that the FMLP responses were because of leukocyte contamination. In contrast, reactive oxygen species production in response to PMA appeared to reflect the oxidant-generating capacity of both leukocytes and spermatozoa.


CONCLUSIONS
The only cells present in the human ejaculate possessing detectable receptors for FMLP and capable of generating reactive oxygen species in response to this reagent come from the leukocyte population. Luminol-dependent, FMLP-induced, chemiluminescence provides a rational basis for monitoring the presence of leukocytes in suspensions of human spermatozoa.
Krausz
C
C
University of Edinburgh, Scotland.
West
K
K
Buckingham
D
D
Aitken
R J
RJ
D002458:Cell Fractionation; D002469:Cell Separation / Q000379:methods*; D005854:Germ Cells / Q000166:cytology; D006801:Humans; D007962:Leukocytes / Q000166:cytology*; D008163:Luminescent Measurements; D008280:Magnetics; D008297:Male; D008722:Methods; D008863:Microspheres; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D012661:Semen / Q000166:cytology* / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D009240:N-Formylmethionine Leucyl-Phenylalanine; D013755:Tetradecanoylphorbol Acetate