Hans Wolff

Comparison of methods to enumerate white blood cells in semen

Seminal WBC counts obtained by an mAb-based immunohistologic method correlated well with seminal granulocyte counts obtained with a simple peroxidase method (rho = +0.70; P < 0.0001). However, total WBC counts were significantly higher than granulocyte counts for most samples. With the immunohistologic method, 17 of 112 samples (15.2%) contained > 10(6) WBC/mL semen, whereas the peroxidase method resulted in only 10 samples (8.9%) with > 10(6) WBC/mL. When the threshold defining leukocytospermia was set at 1 x 10(6) positive cells/mL for both methods, the specificity of the peroxidase test compared with the immunohistology technique was 100% (10/10), but the sensitivity was only 58.8% (10/17). When the threshold for leukocytospermia in the peroxidase test was lowered to 5 x 10(5) positive cells/mL semen, the sensitivity relative to the immunohistology technique increased to 94.1% (16/17), and specificity remained 100% (16/16). Likewise, good interassay sensitivity and specificity values were obtained with thresholds of 10(6) WBC/mL for the peroxidase assay and 2 x 10(6) WBC/mL for the immunohistology assay. We conclude that either peroxidase or immunohistology assays can be used to screen for leukocytospermia, but that more research is needed to establish thresholds for pathological levels of WBC in semen using these two approaches. Total round cell counts are of no value for enumerating WBC in semen.

Leukocytospermia is associated with poor semen quality

regions in the United States, such as high-risk areas of New York City, , infection may be present in more than 3qo of reproductive-age women, most of whom do not know they are infected. Women who either live in areas of the United States in which acquired immunodeficiency syndrome (AIDS) is endemic or are living in other parts of the country and are at high risk for AIDS, yet refuse to be tested, should be counseled regarding the potential for transmission of the virus to uninfected offspring via breast-feeding.

Immunohistologic characterization and quantitation of leukocyte subpopulations in human semen**Supported by the Fearing Research Laboratory Endowment and National Institutes of Health grants AI 23669 and CA 42738.††Presented at the 12th Annual Meeting of the American Society of Andrology, March 6 to 9, 1987, Denver, Colorado.

Various major leukocyte subpopulations (granulocytes, monocytes/macrophages, B lymphocytes, helper and suppressor/cytotoxic T lymphocytes) were detectable in human semen by applying monoclonal antibodies (MAbs) specific for pan-leukocyte and leukocyte subpopulation markers. MAb-labeled cells in smears of washed human semen were visualized by a standard streptavidin-biotin immunoperoxidase method. This approach was validated by testing the MAb panel against sections of human testis and epididymis to rule out cross-reactivity with immature germ cells or other reproductive tract cells, and against peripheral blood leukocytes that had been incubated in human semen to ascertain leukocyte marker stability in seminal plasma. When the immunoperoxidase technique was applied to semen smears from 17 proven fertile men and 51 randomly selected infertility patients, the authors found highly varying total leukocyte numbers, ranging from 8970 to 20,520,000 per ejaculate among fertile men (median, 170,000), and from 43,120 to 104,580,000 among infertility patients (median, 1,035,000). Higher means and medians of granulocytes, monocytes/macrophages, and lymphocyte subsets were seen in the infertility group. These results warrant further investigation on the diagnostic utility of semen leukocyte profiles and the possible role of seminal leukocytes in male infertility.

Evaluation of granulocyte elastase as a seminal plasma marker for leukocytospermia**Supported by the Fearing Research Laboratory Endowment and National Institutes of Health grants AI 23669 and CA42738.

To evaluate the relationship between concentrations of the granulocytic enzyme polymorphonuclear (PMN) elastase in seminal plasma and the presence of high numbers of white blood cells (WBC) in semen, specimens from 118 infertility patients were analyzed for each variable. PMN-elastase levels were determined by an enzyme-linked immunosorbent assay and WBC numbers by a recently described immunohistologic technique. PMN-elastase levels in seminal plasma correlated well with WBC numbers in semen (r = +0.755). A three-step grading system of PMN-elastase values as low/normal (less than 250 ng/ml), intermediate (250 to 1000 ng/ml) and high/pathologic (greater than 1000 ng/ml) showed a highly significant relationship to a similar WBC number grading system: low/normal, less than 10(5) WBC/ml, intermediate, 10(5) to 10(6) WBC/ml, and high/leukocytospermic, greater than 10(6) WBC/ml semen. Incubation of semen at 37 degrees C resulted in a slight continuous decrease of immunoreactive PMN-elastase at a rate of approximately 10% per 3 hours. As all 18 samples with more than 1000 ng PMN-elastase per ml seminal plasma showed more than 10(6) WBC/ml semen, PMN-elastase levels above 1000 ng/ml were diagnostic for leukocytospermia.