Okada H, Hayashi A, Matsumoto O, Kamidono S
Department of Urology, Kobe University School of Medicine.
Nihon Hinyokika Gakkai Zasshi. 1993 Feb;84(2):350-4. doi: 10.5980/jpnjurol1989.84.350.
It is hard to differentiate leucocytes from immature germ cells among many round cells in ejaculated semen. Recently monoclonal antibodies (MoAbs) which recognize surface antigen of various subtypes of leucocytes are available. In this study we applied a panel of MoAbs for leucocyte subpopulations (granulocytes, T-lymphocytes, B-lymphocytes, Monocytes/Macrophages and pan-leucocytes). Semen smears from 64 mild oligozoospermic patients and 5 fertile volunteers were stained by immunohistological technique (avidin-biotin-immunoperoxidase method). We found highly varying total leucocyte numbers ranging from 1200 to 2.2 x 10(8) per ejaculate. When comparing semen from infertile patients with those from fertile donors, statistically significant higher medians of total leucocytes, total granulocytes and monocytes/macrophages were seen in infertile group. We compared a total number of leucocytes obtained from conventional Papanicolaou staining with that from immunohistochemical staining. Both were almost identical but the former gave a little bit higher value, because of the difficulty in differentiating leucocytes from immature germ cell by Papanicolaou staining. Moreover we performed an experiment to evaluate the applicability of flow-cytometry (FCM) to count leucocytes in semen. Cells in the semen smears were stained with MoAbs recognizing the surface antigen shared by every leucocyte, and percentage of the leucocytes which were thus stained with MoAbs was compared with that determined by FCM. Results from FCM constantly gave lower value than these from immunostaining. This is probably because FCM could not differentiate leucocytes from contaminated non cell substances (e.g. cell debris). From the present study it is not adequate to use FCM for the detection of leucocytes in semen as applied in haematology. Further techniques to remove non-cellular substances should be established for its application in semen analysis.
在射出的精液中，众多圆形细胞里很难区分白细胞和未成熟生殖细胞。近来，可获得识别白细胞各亚型表面抗原的单克隆抗体（MoAbs）。在本研究中，我们应用了一组针对白细胞亚群（粒细胞、T淋巴细胞、B淋巴细胞、单核细胞/巨噬细胞和全白细胞）的MoAbs。采用免疫组织学技术（抗生物素蛋白-生物素-免疫过氧化物酶法）对64例轻度少精子症患者和5名有生育能力的志愿者的精液涂片进行染色。我们发现每次射精的白细胞总数差异很大，范围从1200到2.2×10⁸ 。将不育患者的精液与有生育能力供者的精液相比较，不育组的白细胞总数、粒细胞总数和单核细胞/巨噬细胞的中位数在统计学上显著更高。我们比较了常规巴氏染色获得的白细胞总数与免疫组化染色获得的白细胞总数。两者几乎相同，但前者的值略高一点，因为巴氏染色难以区分白细胞和未成熟生殖细胞。此外，我们进行了一项实验以评估流式细胞术（FCM）在精液白细胞计数中的适用性。用识别每种白细胞共有的表面抗原的MoAbs对精液涂片上的细胞进行染色，并将由此被MoAbs染色的白细胞百分比与FCM测定的百分比进行比较。FCM的结果始终低于免疫染色的结果。这可能是因为FCM无法区分白细胞与污染的非细胞物质（如细胞碎片）。从本研究来看，在精液分析中应用时，将FCM用于精液中白细胞的检测并不合适。应建立进一步去除非细胞物质的技术以用于精液分析。
