Batten S J, Osmond D G
J Immunol Methods. 1984 Sep 4;72(2):381-99. doi: 10.1016/0022-1759(84)90007-3.
An in vivo cell surface labelling technique using radioautography has been developed to visualise the distribution of IgM-bearing B lymphocytes within the bone marrow. Anaesthetized 3-week-old mice were perfused via the common iliac artery with: (1) serum-containing medium (SCM), (2) 125I-labelled anti-IgM antibody in SCM, (3) SCM, and (4) fixative. In radioautographic sections of femoral marrow labelled surface IgM+ cells were observed either singly or in small clusters throughout the extravascular haemopoietic marrow cords. Binding specificity was demonstrated by the displacement of 125I-anti-IgM labelling by excess anti-IgM and by the binding of perfused 125I-anti-H-2Kk antibody in CBA/J (H-2Kk) mice but not in C57BL/6 (H-2Kb) mice. Quantitative analysis of radioautographic sections revealed an even distribution of labelled cells throughout CBA/J marrow perfused with 125I-anti-H-2Kk, indicating a uniform accessibility of perfused antibody to cells in the haemopoietic cords. This labelling pattern contrasted with that in 125I-anti-IgM perfused animals in which surface IgM+ cells, although widely distributed in the bone marrow, showed areas of concentration, speculatively clones of maturing B lymphocytes. This method of labelling surface IgM and other cell markers in situ provides an approach to study the microenvironment of B lymphocyte genesis in the bone marrow.
一种利用放射自显影的体内细胞表面标记技术已被开发出来，用于观察骨髓中携带IgM的B淋巴细胞的分布。对3周龄的麻醉小鼠通过髂总动脉灌注：(1)含血清培养基(SCM)，(2)SCM中125I标记的抗IgM抗体，(3)SCM，以及(4)固定剂。在股骨骨髓的放射自显影片中，标记的表面IgM+细胞在血管外造血骨髓索中单独或成小簇出现。通过过量抗IgM取代125I-抗IgM标记以及在CBA/J(H-2Kk)小鼠而非C57BL/6(H-2Kb)小鼠中灌注的125I-抗H-2Kk抗体的结合，证明了结合特异性。对放射自显影片的定量分析显示，在用125I-抗H-2Kk灌注的CBA/J骨髓中，标记细胞均匀分布，表明灌注抗体对造血索中的细胞具有均匀的可及性。这种标记模式与用125I-抗IgM灌注的动物不同，在后者中，表面IgM+细胞虽然广泛分布于骨髓中，但显示出集中区域，推测为成熟B淋巴细胞克隆。这种原位标记表面IgM和其他细胞标志物的方法为研究骨髓中B淋巴细胞发生的微环境提供了一种途径。
