Rosen H, Gordon S
Sir William Dunn School of Pathology, University of Oxford.
Eur J Immunol. 1990 Jun;20(6):1251-8. doi: 10.1002/eji.1830200609.
We have examined the migration of murine macrophages from the vascular compartment to normal and inflammatory tissues by the adoptive transfer of resident peritoneal macrophages (RPM phi) fluorescently labeled with the hydrophobic dye 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI). After initial labeling of the plasma membrane of RPM phi, the dye accumulated stably in intracellular vesicles of low density (rho = 1.042-1.045 kg/l) and cells remained viable in culture for 4 weeks. Like the normal monocyte, DiI-RPM phi, but not exudate-derived or fixed cells, migrated to peritoneal exudates, following i.v. adoptive transfer, by a mechanism inhibitable by an antibody to the type 3 complement receptor. In the absence of an inflammatory stimulus there was no migration to the peritoneal cavity, and DiI-RPM phi accumulated within 4 h in the red pulp and marginal zone of the spleen. By day 6 these cells still formed a tight ring of fluorescence in the marginal zone alone, outside the marginal metallophil cells. DiI-RPM phi injected into the peritoneal cavity migrated to the parathymic lymph nodes where they were found in the subcapsular sinus and in the medullary cords, whereas very few fluorescent cells could be found in the T cell areas. The migration of RPM phi to lymphoid organs required viable cells but, unlike the recruitment of cells to peritoneal exudates, was not inhibitable by antibodies to CR3. We conclude that the RPM phi is a useful surrogate for the analysis of constitutive and induced monocyte migration to secondary lymphoid and inflammatory sites, respectively.
我们通过采用疏水性染料1,1'-二辛基-3,3,3',3'-四甲基吲哚羰花青高氯酸盐(DiI)荧光标记的驻留腹膜巨噬细胞(RPM phi)进行过继转移,研究了小鼠巨噬细胞从血管腔迁移至正常组织和炎症组织的过程。在用DiI对RPM phi的质膜进行初始标记后,染料稳定地积聚在低密度(ρ = 1.042 - 1.045 kg/l)的细胞内囊泡中,并且细胞在培养中可存活4周。与正常单核细胞一样,DiI - RPM phi而非渗出液来源的细胞或固定细胞,在静脉内过继转移后,通过一种可被抗3型补体受体抗体抑制的机制迁移至腹膜渗出液。在没有炎症刺激的情况下,不会向腹膜腔迁移,并且DiI - RPM phi在4小时内积聚在脾脏的红髓和边缘区。到第6天时,这些细胞仍仅在边缘区形成一个紧密的荧光环,位于边缘嗜金属细胞之外。注入腹膜腔的DiI - RPM phi迁移至胸腺旁淋巴结,在那里它们存在于被膜下窦和髓索中,而在T细胞区域中很少能发现荧光细胞。RPM phi向淋巴器官的迁移需要活细胞,但与细胞募集至腹膜渗出液不同,它不受抗CR3抗体的抑制。我们得出结论,RPM phi分别是分析组成型和诱导型单核细胞向次级淋巴组织和炎症部位迁移的有用替代物。