Indirect immunofluorescence using monoclonal antibodies for the detection of leukocytospermia: comparison with peroxidase staining.
The presence of increased number of leukocytes in semen is indicative of inflammation in the male genital tract. Inflammatory processes at this level may lead to marked impairment of sperm function, and finally to a reduction in their fertilizing capability. An immunocytological technique for the detection of seminal leukocytes was evaluated in this study. As part of the standardization technique, different fixation methods were tested to ascertain whether samples could be stored and examined later. It was found that fixation with cold acetone at freezing temperatures retained immunoreactivity until day 11 of storage. All other methods showed a significant loss of immunoreactivity, from as little as day 2 of storage. In 46 specimens with elevated numbers of round cells, number of peroxidase-positive cells and number and type of leukocytes were evaluated by means of indirect immunofluorescence. Determination of peroxidase-positive cells to detect leukocytospermia, the standard procedure recommended by the WHO, was compared with the indirect immunofluorescence technique using monoclonal antibodies. While 19 of 46 patients showed high numbers of leukocytes in the ejaculate, as determined by the immunocytological method, only 9 of these were identified to be leukocytospermic, according to the WHO (standard) procedure. This difference was statistically significant (P < 0.01) and indicates that the standard method of detection of seminal leukocytes may be inaccurate.
Villegas
J
J
Department of Medicine, Universidad de La Frontera, Temuco, Chile.
Schulz
M
M
Vallejos
V
V
Henkel
R
R
Miska
W
W
Sánchez
R
R
D000096:Acetone; D000911:Antibodies, Monoclonal / Q000276:immunology; D015415:Biomarkers; D000431:Ethanol; D005404:Fixatives; D019084:Fluorescent Antibody Technique, Indirect; D006098:Granulocytes / Q000166:cytology* / Q000276:immunology; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis; D007962:Leukocytes / Q000166:cytology* / Q000276:immunology; D016256:Lewis X Antigen / Q000032:analysis; D008297:Male; D009195:Peroxidase; D012661:Semen / Q000166:cytology* / Q000276:immunology; D013194:Staining and Labeling / Q000379:methods
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D005404:Fixatives; D016256:Lewis X Antigen; D000096:Acetone; D000431:Ethanol; D009195:Peroxidase; D017493:Leukocyte Common Antigens