Crago S S, Prince S J, Pretlow T G, McGhee J R, Mestecky J
Clin Exp Immunol. 1979 Dec;38(3):585-97.
Analyses of the cells present in human colostrum obtained from fifty-four healthy donors during the first four days of lactation revealed that there were 3.3 x 10(6) (range 1.1 x 10(5)--1.2 x 10(7)) cells per ml of colostrum. Based on histochemical examinations, it was found that this population consisted of 30--47% macrophages, 40--60% polymorphonuclear leucocytes, 5.2--8.9% lymphocytes, and 1.3--2.8% colostral corpuscles; epithelial cells were rarely encountered. The identity of various cell types was confirmed by Wright's stain and by a series of histochemical techniques which disclosed the presence of non-specific esterase, peroxidase, and lipids. For further characterization, the different types of cells were separated by various methods, such as Ficoll-Hypaque density centrifugation, isokinetic centrifugation on a linear Ficoll gradient, adherence to glass or plastic, and phagocytosis of carbonyl iron. Immunohistochemical staining with FITC- and/or TRITC-labelled reagents to IgA, IgM, IgG, K- and lambda-chains, secretory component, lactoferrin, and alpha-lactalbumin were applied to unseparated as well as separated colostral cells. Polymorphonuclear leucocytes (staining for peroxidase) as well as macrophages and colostral corpuscles (staining for non-specific esterase) exhibited numerous intracellular vesicles that contained lipids as well as various combinations of milk proteins. Lymphoid cells did not stain with any of these reagents and plasma cells were not detected among the colostral cells. Individual phagocytic cells contained immunoglobulins of the IgA and IgM classes, both K and lambda light chains, secretory component, lactoferrin, and alpha-lactalbumin. The coincidental appearance of these proteins in single, phagocytic cells but not in lymphoid cells indicate that the cells acquired these proteins by ingestion from the environment. Markers commonly used for the identification of B lymphocytes (surface immunoglobulins) and T lymphocytes (receptors for sheep red blood cells) were unreliable for the analysis of colostral cells (unless accompanied by subsequent morphological characterization) because strong fluorescence was observed on the surface of many non-lymphoid cells and because numerous macrophages and colostral corpuscles formed rosettes with sheep red blood cells (SRBC). Lymphocytes, often found in association with colostral macrophages or corpuscles, were classified as T cells.
对54名健康捐赠者在哺乳期头四天采集的人初乳中的细胞进行分析发现，每毫升初乳中有3.3×10⁶个细胞（范围为1.1×10⁵ - 1.2×10⁷个）。基于组织化学检查，发现该细胞群体由30% - 47%的巨噬细胞、40% - 60%的多形核白细胞、5.2% - 8.9%的淋巴细胞和1.3% - 2.8%的初乳小体组成；很少遇到上皮细胞。通过瑞氏染色和一系列组织化学技术证实了各种细胞类型的身份，这些技术揭示了非特异性酯酶、过氧化物酶和脂质的存在。为了进一步表征，通过多种方法分离不同类型的细胞，如Ficoll - Hypaque密度离心、在线性Ficoll梯度上的等速离心、贴壁于玻璃或塑料以及吞噬羰基铁。用异硫氰酸荧光素（FITC）和/或四甲基异硫氰酸罗丹明（TRITC）标记的试剂对IgA、IgM、IgG、κ和λ链、分泌成分、乳铁蛋白和α - 乳白蛋白进行免疫组织化学染色，应用于未分离以及已分离的初乳细胞。多形核白细胞（过氧化物酶染色）以及巨噬细胞和初乳小体（非特异性酯酶染色）表现出许多含有脂质以及各种乳蛋白组合的细胞内小泡。淋巴细胞对这些试剂均无染色，且在初乳细胞中未检测到浆细胞。单个吞噬细胞含有IgA和IgM类免疫球蛋白、κ和λ轻链、分泌成分、乳铁蛋白和α - 乳白蛋白。这些蛋白质在单个吞噬细胞中同时出现而在淋巴细胞中未出现，表明细胞是通过从环境中摄取而获得这些蛋白质的。通常用于鉴定B淋巴细胞（表面免疫球蛋白）和T淋巴细胞（绵羊红细胞受体）的标志物对于初乳细胞分析不可靠（除非随后进行形态学表征），因为在许多非淋巴细胞表面观察到强荧光，并且因为许多巨噬细胞和初乳小体与绵羊红细胞形成花环。经常与初乳巨噬细胞或小体相关联发现的淋巴细胞被分类为T细胞。
