Kucharova Karolina, Stallcup William B
Cancer Center, Tumor Microenvironment and Cancer Immunology Program, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California, United States of America.
PLoS One. 2017 Nov 2;12(11):e0187530. doi: 10.1371/journal.pone.0187530. eCollection 2017.
We used a bone marrow transplantation approach to distinguish the activities of bone marrow-derived macrophages from the activities of central nervous system-resident microglia in phenomena associated with axon demyelination and remyelination. We transplanted wild type or germline NG2 null beta-actin-EGFP expressing bone marrow into irradiated wild type or NG2 null recipient mice, followed by analysis of lysolecithin-induced spinal cord demyelination and remyelination and quantification of Iba-1+/ F4/80+/ EGFP+ macrophages and Iba-1+/ F4/80+/ EGFP- microglia. One week after microinjection of 1% lysolecithin into the spinal cord, wild type recipients receiving NG2 null bone marrow exhibit greatly reduced infiltration of macrophages into lesions, compared to wild type recipients receiving wild type bone marrow. Wild type bone marrow recipients also exhibit larger numbers of demyelinated axons than NG2 null recipients, indicative of macrophage participation in the initial myelin damage. However, wild type bone marrow recipients also exhibit superior myelin repair at 6 weeks post-injury, compared to NG2 null bone marrow recipients, demonstrating the additional importance of macrophages in remyelination. Incompletely repaired lesions in NG2 null bone marrow recipients at 6 weeks post-injury retain elevated numbers of macrophages, in contrast to lower numbers of macrophages in more completely repaired lesions in wild type bone marrow recipients. This suggests that NG2 expression renders macrophages more effective in myelin repair and less likely to promote chronic inflammation. Effective macrophage involvement in myelin repair is due in part to effects on the proliferation and/or recruitment of oligodendrocyte progenitor cells. Reduced numbers of oligodendrocyte progenitors are seen in lesions in NG2 null bone marrow recipients, likely due to deficits in macrophage production of oligodendrocyte progenitor-relevant mitogens and in phagocytosis of inhibitory myelin debris. Microglia also appear to be important for clearance of myelin debris, as indicated by reduced phagocytosis in NG2 null recipients receiving wild type bone marrow.
我们采用骨髓移植方法,以区分在轴突脱髓鞘和髓鞘再生相关现象中,骨髓来源的巨噬细胞与中枢神经系统驻留的小胶质细胞的活性。我们将表达野生型或种系NG2缺失的β-肌动蛋白-EGFP的骨髓移植到经照射的野生型或NG2缺失的受体小鼠中,随后分析溶血卵磷脂诱导的脊髓脱髓鞘和髓鞘再生情况,并对Iba-1+/F4/80+/EGFP+巨噬细胞和Iba-1+/F4/80+/EGFP-小胶质细胞进行定量分析。在脊髓内显微注射1%溶血卵磷脂一周后,与接受野生型骨髓的野生型受体相比,接受NG2缺失骨髓的野生型受体损伤部位巨噬细胞浸润显著减少。野生型骨髓受体的脱髓鞘轴突数量也比NG2缺失受体多,这表明巨噬细胞参与了最初的髓鞘损伤。然而,与NG2缺失骨髓受体相比,野生型骨髓受体在损伤后6周时也表现出更好的髓鞘修复,这表明巨噬细胞在髓鞘再生中具有额外的重要性。与野生型骨髓受体中修复更完全的损伤部位巨噬细胞数量较少相比,NG2缺失骨髓受体在损伤后6周时未完全修复的损伤部位巨噬细胞数量仍然较高。这表明NG2的表达使巨噬细胞在髓鞘修复中更有效,并且不太可能促进慢性炎症。巨噬细胞有效参与髓鞘修复部分归因于其对少突胶质前体细胞增殖和/或募集的影响。在NG2缺失骨髓受体的损伤部位可见少突胶质前体细胞数量减少,这可能是由于巨噬细胞产生少突胶质前体相关促有丝分裂原的缺陷以及对抑制性髓鞘碎片的吞噬作用不足所致。小胶质细胞似乎对髓鞘碎片的清除也很重要,这在接受野生型骨髓的NG2缺失受体中吞噬作用降低得到了体现。