Komai-Koma M, Wilkinson P C
Department of Immunology, University of Glasgow (Western Infirmary), UK.
Immunology. 1997 Jan;90(1):23-9. doi: 10.1046/j.1365-2567.1997.d01-2131.x.
The locomotor properties of B cells isolated from the germinal centres (GC) of human tonsils were studied using polarization, collagen gel invasion and micropore filter assays. The proportion of motile GC cells in the freshly isolated population was small. During culture in interleukin-4 (IL-4)+anti-CD40, but not in control medium, the proportion of polarized cells increased and these cells migrated actively into collagen gels. After 24 hr culture, most of the surviving population was in locomotor morphology. The locomotor population consisted mainly of centrocytes in the G1 phase of growth. More locomotor cells than spherical cells took up [3H]uridine, but locomotor cells did not take up [3H]thymidine. After culture for 6 hr in IL-4+anti-CD40, GC B cells were tested in short-term polarization assays and filter assays for their response to chemoattractants. In both assays, a proportion of the cells responded to anti-IgA and to anti-IgA F(ab')2 fragments at 1 ng/ml., or to anti-IgG, anti-IgM and F(ab')2 fragments of these antibodies at 100 ng-1 microgram/ml. A checkerboard filter assay showed a good chemokinetic response and a weaker chemotactic response of GC cells to anti-IgA. Expression of Fc gamma RII (CD32) was increased after culture in IL-4+anti-CD40, and these cultured cells responded in filter and polarization assays to anti-CD32. Thus culture in IL-4 and anti-CD40 not only rescues GC B cells, but also increases their locomotor capacity and allows them to respond in chemotaxis assays to anti-immunoglobulin.
利用极化、胶原凝胶侵袭和微孔滤膜试验，研究了从人扁桃体生发中心（GC）分离的B细胞的运动特性。在新鲜分离的细胞群体中，具有运动能力的GC细胞比例较小。在白细胞介素-4（IL-4）+抗CD40培养基中培养时，极化细胞的比例增加，且这些细胞能活跃地迁移到胶原凝胶中，而在对照培养基中培养时则无此现象。培养24小时后，大多数存活的细胞群体呈现出运动形态。运动细胞群体主要由处于生长G1期的中心细胞组成。运动细胞比球形细胞摄取更多的[3H]尿苷，但运动细胞不摄取[3H]胸苷。在IL-4+抗CD40中培养6小时后，对GC B细胞进行短期极化试验和滤膜试验，检测它们对趋化因子的反应。在这两种试验中，一部分细胞对1 ng/ml的抗IgA和抗IgA F(ab')2片段有反应，或对100 ng - 1 μg/ml的这些抗体的抗IgG、抗IgM和F(ab')2片段有反应。棋盘式滤膜试验显示GC细胞对抗IgA有良好的化学动力学反应和较弱的趋化反应。在IL-4+抗CD40中培养后，FcγRII（CD32）的表达增加，且这些培养细胞在滤膜试验和极化试验中对抗CD32有反应。因此，在IL-4和抗CD40中培养不仅能挽救GC B细胞，还能增加它们的运动能力，并使它们在趋化试验中对抗免疫球蛋白产生反应。
