Fathy A, Chen S J, Novak N, Schuppe H-C, Haidl G, Allam J-P
Department of Dermatology and Allergy/Andrology Unit, Rheinische Friedrich-Wilhelms University, Bonn, Germany.
Andrologia. 2014;46(9):1004-12. doi: 10.1111/and.12188. Epub 2013 Nov 8.
Genital tract inflammation is considered as a major cause of male infertility with leucocytospermia as widely used diagnostic marker. However, threshold of 10(6) leucocytes ml(-1) recommended by the WHO is a matter of debate. Moreover, leucocyte subpopulations and their impact cannot be identified by the routine peroxidase method (POM). Ejaculates of subfertile men (n = 47) were analysed by flow cytometry (FACS) using a bead-based method. Leucocytes were identified by CD18 and further divided into macrophages (HLA-Dr+/CD66abce-) and neutrophils (HLA-Dr-/CD66abce+). IL-1β, TNF-α and IL-6 production was investigated in these subpopulations. It was found that CD18-positive cells correlated significantly with POM. However, only in samples with POM below 10(6) per millilitre, FACS detected significantly higher leucocyte numbers. Moreover, in 31% of these samples, FACS leucocyte detection reached threshold values greater than 1 × 10(6) ml(-1) , fulfilling the criteria for diagnosis of leucocytospermia. Neutrophils were the predominating leucocyte population. Nevertheless, in 24% of samples, macrophages encountered more than 50% of leucocytes. Most interestingly, only macrophages produced significant amounts of IL-1β, TNF-α and IL-6. It is concluded that FACS improves detection and functional differentiation of seminal leucocytes as one of the diagnostic hallmarks of male genital tract inflammation.
生殖道炎症被认为是男性不育的主要原因，白细胞精子症是广泛使用的诊断标志物。然而，世界卫生组织推荐的每毫升10⁶个白细胞的阈值存在争议。此外，常规过氧化物酶法（POM）无法识别白细胞亚群及其影响。采用基于微珠的方法，通过流式细胞术（FACS）对47例不育男性的精液进行分析。通过CD18识别白细胞，并进一步分为巨噬细胞（HLA-Dr⁺/CD66abce⁻）和中性粒细胞（HLA-Dr⁻/CD66abce⁺）。研究了这些亚群中白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的产生情况。发现CD18阳性细胞与POM显著相关。然而，只有在每毫升POM低于10⁶的样本中，FACS检测到的白细胞数量显著更高。此外，在这些样本的31%中，FACS白细胞检测达到大于1×10⁶每毫升的阈值，符合白细胞精子症的诊断标准。中性粒细胞是主要的白细胞群体。然而，在24%的样本中，巨噬细胞占白细胞的比例超过50%。最有趣的是，只有巨噬细胞产生大量的IL-1β、TNF-α和IL-6。结论是，作为男性生殖道炎症诊断标志之一，FACS改善了精液白细胞的检测和功能分化。
