Petrasch S, Perez-Alvarez C, Schmitz J, Kosco M, Brittinger G
Department of Hematology, University of Essen, FRG.
Eur J Immunol. 1990 May;20(5):1013-8. doi: 10.1002/eji.1830200510.
Follicular dendritic cells (FDC) are located within follicles of secondary lymphoid tissue and in lymph nodes of patients with germinal center cell-derived non-Hodgkin lymphomas. Reliable antigenic phenotyping of FDC within tissue sections has been difficult due to simultaneous labeling of the surrounding germinal center cells. Using an enzyme cocktail to digest human tonsils and cervical lymph nodes with subsequent fractionation by albumin gradient centrifugation, cell isolates containing up to 20% FDC were obtained. This preparation allowed the determination of antigenic phenotype on individual FDC. Molecules expressed by FDC were detected by an isotype-specific immunocytochemical double-labeling procedure, i.e. a monoclonal antibody (mAb) specific for FDC (KiM4 or DRC1) in conjunction with a mAb reactive against an additional antigenic determinant. Nonspecific binding of mAb to immunoglobulin Fc receptors located on FDC membranes was avoided by incubation of cells with human IgG aggregates prior to immunostaining. The results revealed that isolated FDC from these lymphoid tissues express transferrin receptors, the intercellular adhesion molecule 1, class II antigens, the B cell antigens CD20 and CD21, and the myelomonocytic properties CD11b and CD14. Immunoglobulin mu or gamma heavy chains and the B cell antigens CD23 and CD24 are detected on 50% of an isolated FDC population. These FDC are negative for the T helper cell antigen CD4, the B cell cell antigens CD19 and CD22, the immunolobulin alpha and delta chains and the S-100 protein. FDC isolated from lymph nodes of patients with low-grade malignant non-Hodgkin lymphoma, identified by DRC1 or KiM4 mAb, presented the same antigenic profile as seen on FDC from nonmalignant tissue. This suggests that FDC from lymphoma tissue isolated in this manner have the same properties as those found in normal tissue.
滤泡树突状细胞(FDC)位于次级淋巴组织的滤泡内以及生发中心细胞来源的非霍奇金淋巴瘤患者的淋巴结中。由于周围生发中心细胞的同时标记,在组织切片中对FDC进行可靠的抗原表型分析一直很困难。使用酶混合物消化人扁桃体和颈部淋巴结,随后通过白蛋白梯度离心进行分级分离,获得了含有高达20% FDC的细胞分离物。这种制备方法能够确定单个FDC的抗原表型。通过同型特异性免疫细胞化学双标记程序检测FDC表达的分子,即针对FDC的单克隆抗体(mAb)(KiM4或DRC1)与针对另一种抗原决定簇的反应性mAb结合使用。在免疫染色之前,通过用人IgG聚集体孵育细胞来避免mAb与位于FDC膜上的免疫球蛋白Fc受体的非特异性结合。结果显示,从这些淋巴组织中分离出的FDC表达转铁蛋白受体、细胞间粘附分子1、II类抗原、B细胞抗原CD20和CD21,以及髓单核细胞特性CD11b和CD14。在50%的分离FDC群体中检测到免疫球蛋白μ或γ重链以及B细胞抗原CD23和CD24。这些FDC对T辅助细胞抗原CD4、B细胞抗原CD19和CD22、免疫球蛋白α和δ链以及S-100蛋白呈阴性。通过DRC1或KiM4 mAb鉴定的从低度恶性非霍奇金淋巴瘤患者淋巴结中分离出的FDC,呈现出与非恶性组织中FDC相同的抗原谱。这表明以这种方式从淋巴瘤组织中分离出的FDC具有与正常组织中发现的FDC相同的特性。