Kroese F G, Wubbena A S, Opstelten D, Deenen G J, Schwander E H, De Leij L, Vos H, Poppema S, Volberda J, Nieuwenhuis P
Eur J Immunol. 1987 Jul;17(7):921-8. doi: 10.1002/eji.1830170705.
Three mouse monoclonal antibodies (mAb) directed against rat B lineage antigens were produced. The mAb, designated HIS14 (IgG1), HIS22 (IgM) and HIS24 (IgG2b), were characterized for binding to lymphoid and nonlymphoid tissues by immunoperoxidase staining of frozen sections and by (double-) immunofluorescence staining of single cell suspensions from lymphoid organs. HIS14 recognized a pan B cell determinant: it reacted with virtually all cells of each anatomic B cell compartment and with about 95% of surface (s)Ig+ cells in thoracic duct lymph and in suspensions of spleen and lymph nodes. HIS22 and HIS24 detected B lineage-associated antigens expressed by major subpopulations of B cells. HIS22 predominantly stained the lymphocyte corona, but not (or weakly) the germinal centers and splenic marginal zones, whereas HIS24 reacted with both corona and germinal center and not (or weakly) with marginal zone. In accordance with this, substantial proportions of sIg+ cells in spleen cell suspensions did not express HIS22 or HIS24 determinants (20% and 27%, respectively). In bone marrow the vast majority of cytomplasmic mu+ pre-B cells were HIS14+ and HIS24+, and up to one third also HIS22+, indicating an appearance of the determinants early in B lymphocytopoiesis. The antigens recognized by HIS14, HIS22 and HIS24 are lost during the final stage of B cell differentiation: none of the mAb bound to plasma cells. As far as detectable, neither cells of myeloid and erythroid lineages in bone marrow nor thymocytes were stained by HIS14, HIS22, or HIS24. In suspensions of peripheral lymphoid organs (spleen and lymph nodes) but not in thoracic duct lymph, HIS14 and HIS24 labeled a small proportion (12% and 14%, respectively) of Ig- cells. HIS22 did not bind to Ig- peripheral lymphocytes. Reactivity of HIS14, HIS22 and HIS24 with nonlymphoid tissues was virtually absent; HIS22 stained the high endothelial venules in lymph nodes and Peyer's patches. As determined by immunoblotting, the antigenic determinants on lymph node cells recognized by HIS14, HIS22 and HIS24 were present on molecules with an apparent molecular mass of 205 kDa, 210 (and 175) kDa and 205 kDa, respectively, which is similar to the molecular mass of the B cell form of the rat leukocyte common antigen. In addition, the antigens recognized by HIS14, HIS22 and HIS24 co-capped with the leukocyte common antigen. This suggests that each of the three mAb recognize determinants present on the B cell form of the leukocyte common antigen.
制备了三种针对大鼠B系抗原的小鼠单克隆抗体（mAb）。这些单克隆抗体分别命名为HIS14（IgG1）、HIS22（IgM）和HIS24（IgG2b），通过对冰冻切片进行免疫过氧化物酶染色以及对来自淋巴器官的单细胞悬液进行（双重）免疫荧光染色，来鉴定它们与淋巴组织和非淋巴组织的结合情况。HIS14识别一种泛B细胞决定簇：它与每个解剖学B细胞区室的几乎所有细胞发生反应，并且与胸导管淋巴以及脾脏和淋巴结悬液中约95%的表面（s）Ig⁺细胞发生反应。HIS22和HIS24检测到B细胞主要亚群表达的B系相关抗原。HIS22主要染色淋巴细胞冠，但不（或弱）染色生发中心和脾边缘区，而HIS24与冠和生发中心都发生反应，与边缘区不（或弱）反应。据此，脾细胞悬液中相当比例的sIg⁺细胞不表达HIS22或HIS24决定簇（分别为20%和27%）。在骨髓中，绝大多数细胞质μ⁺前B细胞是HIS14⁺和HIS24⁺，高达三分之一也是HIS22⁺，这表明这些决定簇在B淋巴细胞生成早期就出现了。HIS14、HIS22和HIS24识别的抗原在B细胞分化的最后阶段丢失：没有一种单克隆抗体与浆细胞结合。就可检测到的情况而言，骨髓中的髓系和红系细胞以及胸腺细胞均未被HIS14、HIS22或HIS24染色。在周围淋巴器官（脾脏和淋巴结）的悬液中，但不在胸导管淋巴中，HIS14和HIS24标记了一小部分（分别为12%和14%）Ig⁻细胞。HIS22不与外周Ig⁻淋巴细胞结合。HIS14、HIS22和HIS24与非淋巴组织几乎没有反应性；HIS22染色淋巴结和派伊尔结中的高内皮小静脉。通过免疫印迹法测定，HIS14、HIS22和HIS24识别的淋巴结细胞上的抗原决定簇分别存在于表观分子量为205 kDa、210（和175）kDa以及205 kDa的分子上，这与大鼠白细胞共同抗原的B细胞形式的分子量相似。此外，HIS14、HIS22和HIS24识别的抗原与白细胞共同抗原共帽化。这表明这三种单克隆抗体各自识别存在于白细胞共同抗原的B细胞形式上的决定簇。
