Nibbering P H, Leijh P C, van Furth R
Department of Infectious Diseases, University Hospital, Leiden, The Netherlands.
Immunology. 1987 Oct;62(2):171-6.
The purpose of the present study was to compare the monoclonal antibody (Mab) binding patterns of various tissue macrophages with each other and with blood monocytes. To allow recovery from the effects of the isolation procedure, or to obtain purified populations, macrophages were cultured for 24 hr and 48 hr. For comparison, blood monocytes were also cultured for 24 hr and 48 hr. Mab binding to individual cells, detected by the biotin avidin immunoperoxidase method, was quantified cytophotometrically and the results expressed as the median of the specific mean absorbance per 0.25 micron2 cell surface area or as specific integrated absorbance per cell. Analysis of the quantitative data in relation to the results of subjective evaluation of the peroxidase reaction product, demonstrating Mab binding to cells, yielded three classes for description of the intensity of antigen expression by cells: weak (specific mean absorbance per unit cell surface less than 0.07), moderate (values between 0.07 and 0.14), and intense (values more than 0.14). No matter how the results were expressed, comparison of the Mab binding patterns of macrophages with those of blood monocytes showed that spleen macrophages bound significantly less F4/80 and more M5/114 (Ia antigen). Kupffer cells and skin macrophages bound either approximately the same amount or considerably less of the various Mabs than monocytes did. Pulmonary tissue and alveolar macrophages bound significantly more 30.G.12 (leucocyte antigen), M3/38 (Mac-2 antigen), and M3/84 (Mac-3 antigen) and comparable amounts or considerably less of the other Mabs than the monocytes did. Peritoneal macrophages bound significantly more F4/80, M1/70 (complement receptor III), and 2.4.G.2. (Fc receptor II) and comparable amounts or considerably less of the other Mabs than monocytes did. It is concluded that macrophages from different organs and different anatomical sites within one organ differ from one another, for example, peritoneal macrophages do not resemble any other population of macrophages and alveolar macrophages do not resemble pulmonary tissue macrophages, and differentiation of blood monocytes into tissue macrophages does not show a distinct pattern.
本研究的目的是比较各种组织巨噬细胞之间以及它们与血液单核细胞的单克隆抗体(Mab)结合模式。为了从分离程序的影响中恢复或获得纯化的细胞群体,将巨噬细胞培养24小时和48小时。作为对照,血液单核细胞也培养24小时和48小时。采用生物素抗生物素蛋白免疫过氧化物酶法检测Mab与单个细胞的结合,通过细胞光度法进行定量,结果以每0.25平方微米细胞表面积的特异性平均吸光度中位数或每个细胞的特异性积分吸光度表示。根据过氧化物酶反应产物的主观评估结果对定量数据进行分析,以证明Mab与细胞的结合,得出三类用于描述细胞抗原表达强度的结果:弱(每单位细胞表面积的特异性平均吸光度小于0.07)、中度(值在0.07至0.14之间)和强(值大于0.14)。无论结果如何表示,巨噬细胞与血液单核细胞的Mab结合模式比较表明,脾巨噬细胞结合的F4/80显著较少,而M5/114(Ia抗原)较多。枯否细胞和皮肤巨噬细胞与单核细胞相比,结合的各种Mab量大致相同或显著较少。肺组织和肺泡巨噬细胞结合的30.G.12(白细胞抗原)、Mз/з8(Mac-2抗原)和Mз/84(Mac-3抗原)显著较多,而其他Mab的结合量与单核细胞相当或显著较少。腹腔巨噬细胞结合的F4/80、M1/70(补体受体III)和2.4.G.2.(Fc受体II)显著较多,而其他Mab的结合量与单核细胞相当或显著较少。得出的结论是,来自不同器官以及一个器官内不同解剖部位的巨噬细胞彼此不同,例如,腹腔巨噬细胞与其他任何巨噬细胞群体都不相似,肺泡巨噬细胞与肺组织巨噬细胞也不相似,并且血液单核细胞向组织巨噬细胞的分化没有呈现出明显的模式。