Crocker P R, Gordon S
J Exp Med. 1985 Sep 1;162(3):993-1014. doi: 10.1084/jem.162.3.993.
In situ studies with the mouse macrophage (M phi)-specific antibody, F4/80, have shown that resident M phi in femoral bone marrow (RBMM) form hematopoietic islands with immature myelomonocytic and erythroid cells (Hume, D. A., et al. 1983. J. Exp. Med. 158: 1522). We have isolated these islands (clusters) by collagenase digestion, purified them from single cells by velocity sedimentation, and analyzed their cellular content. The clusters, ranging from 5- to 100 cells, constituted approximately 7% of the total nucleated cells, and greater than 70% contained at least one strongly staining, F4/80+ central M phi. In comparison, less than 26% showed reactivity for alkaline phosphatase, a marker of fibroblastoid reticulum cells. Compared with the nonclustering population, clusters were enriched with RBMM, fibroblastoid cells, and immature hematopoietic cells, but depleted of mature granulocytes and erythrocytes. The RBMM population was purified from other cells in clusters by selective adherence to glass and was compared with resident peritoneal M phi (RPM) for morphology and the presence of antigens, receptors, and enzymes. RBMM spread more extensively than RPM and frequently extended delicate plasma membrane processes. These and subsequent differences were not attributable to the collagenase treatment. Both M phi populations stained positively with antibodies F4/80 and 2.4G2 (Fc receptor IgG1/2b), bore mannosyl/fucosyl receptors, and showed reactivity for acid phosphatase and nonspecific esterase I. In contrast to RPM, RBMM had no detectable Mac-1 antigen (CR3) or complement receptors, but bore higher levels of Fc receptors (IgG2a and IgG2b) and Ia antigens. In addition, RBMM possessed a novel hemagglutinin activity for unopsonized sheep erythrocytes, which was not present on RPM. RBMM showed no respiratory burst activity in response to zymosan particles, but ingested them avidly. The growth properties of clustering and nonclustered populations were compared by measurement of [3H]thymidine incorporation and progenitor assays. Cells in clusters incorporated three- to fourfold more thymidine than nonclustered cells even in the absence of exogenous growth factors, and autoradiography demonstrated that RBMM made contact with proliferating cells. In contrast, the clusters contained over threefold fewer granulocyte/M phi progenitors compared with nonclustering cells. When clusters were cultivated for up to 3 d, there was rapid outgrowth of monocytes and fibroblastoid cells. These studies demonstrate that RBMM bear a distinct morphology and phenotype.(ABSTRACT TRUNCATED AT 400 WORDS)
用小鼠巨噬细胞（M phi）特异性抗体F4/80进行的原位研究表明，股骨骨髓中的驻留M phi（RBMM）与未成熟的骨髓单核细胞和红系细胞形成造血岛（休姆，D.A.等人，1983年，《实验医学杂志》158:1522）。我们通过胶原酶消化分离出这些岛（簇），通过速度沉降从单个细胞中纯化它们，并分析其细胞成分。这些簇由5到100个细胞组成，约占总核细胞的7%，超过70%的簇至少含有一个强染色的F4/80+中央M phi。相比之下，不到26%的簇对碱性磷酸酶有反应，碱性磷酸酶是成纤维细胞样网状细胞的标志物。与非聚集群体相比，簇中富含RBMM、成纤维细胞样细胞和未成熟造血细胞，但成熟粒细胞和红细胞减少。通过选择性贴壁于玻璃，从簇中的其他细胞中纯化出RBMM群体，并与驻留腹膜M phi（RPM）比较形态以及抗原、受体和酶的存在情况。RBMM比RPM铺展得更广泛，并且经常伸出纤细的质膜突起。这些以及随后的差异并非归因于胶原酶处理。两种M phi群体都用抗体F4/80和2.4G2（Fc受体IgG1/2b）呈阳性染色，带有甘露糖基/岩藻糖基受体，并对酸性磷酸酶和非特异性酯酶I有反应。与RPM相反，RBMM没有可检测到的Mac-1抗原（CR3）或补体受体，但带有更高水平的Fc受体（IgG2a和IgG2b）和Ia抗原。此外，RBMM对未调理的绵羊红细胞具有一种新的血凝活性，而RPM上没有这种活性。RBMM对酵母聚糖颗粒没有呼吸爆发活性，但能大量吞噬它们。通过测量[3H]胸苷掺入和祖细胞测定来比较聚集和非聚集群体的生长特性。即使在没有外源性生长因子的情况下，簇中的细胞掺入的胸苷比非聚集细胞多三到四倍，放射自显影显示RBMM与增殖细胞接触。相比之下，与非聚集细胞相比，簇中粒细胞/M phi祖细胞少三倍以上。当簇培养长达3天时，单核细胞和成纤维细胞样细胞迅速生长。这些研究表明RBMM具有独特的形态和表型。（摘要截断于400字）
