Coates P Toby H, Barratt-Boyes Simon M, Zhang Linyou, Donnenberg Vera S, O'Connell Peta J, Logar Alison J, Duncan F Jason, Murphey-Corb Michael, Donnenberg Albert D, Morelli Adrian E, Maliszewski Charles R, Thomson Angus W
Thomas E. Starzl Transplantation Institute, Pittsburgh, PA, USA.
Blood. 2003 Oct 1;102(7):2513-21. doi: 10.1182/blood-2002-09-2929. Epub 2003 Jun 26.
We provide phenotypic and functional evidence of premonocytoid dendritic cells (DCs) and preplasmacytoid DCs in blood and of corresponding DC subsets in secondary lymphoid tissue of rhesus monkeys. Subsets were identified and sorted by 4-color flow cytometry using antihuman monoclonal antibodies cross-reactive with rhesus monkey. To mobilize pre-DC subsets, fms-like tyrosine 3 kinase ligand (Flt3L; 100 microg/kg subcutaneously) was administered for 10 days. Presumptive pre-DC subsets were identified within the lineage- (Lin-) major histocompatibility complex (MHC) class II+ fraction of blood mononuclear cells. Premonocytoid DCs were CD11c+CD123- (interleukin-3Ralpha- [IL-3Ralpha-]). Preplasmacytoid DCs were characterized as CD11c-CD123++ Flt3L increased the CD11c+ pre-DC (7-fold) and CD123++ pre-DC subsets (3-fold) in blood. The freshly isolated CD11c+ pre-DC subset induced modest proliferation of naive allogeneic T cells. After overnight culture with granulocyte macro-phage-colony-stimulating factor (GMCSF) and CD40L, both subsets up-regulated surface costimulatory molecules, and CD11c+ pre-DCs became potent allostimulators. Freshly isolated CD123++ pre-DCs showed typical plasmacytoid morphology and, when cultured with IL-3 and CD40L for 72 hours, developed mature DC morphology. Following stimulation with CD40L, CD11c+ pre-DCs secreted increased levels of IL-12p40. Importantly, herpes simplex virus-stimulated CD123++ pre-DCs, but not CD11c+ pre-DCs, secreted interferon-alpha (IFN-alpha). Corresponding DC subsets were identified by flow analysis and immunohistochemistry in lymph nodes wherein both populations were increased 2- to 3-fold by Flt3L administration. CD123+ pre-DCs produced IFN-alpha in response to in vivo viral infection. Thus, rhesus monkeys exhibit 2 distinct DC precursor populations that closely resemble those of humans. Both are mobilized into blood and lymphoid tissue by Flt3L, offering potential for their further characterization and possible therapeutic application.
我们提供了恒河猴血液中前单核细胞样树突状细胞（DCs）和前浆细胞样DCs以及二级淋巴组织中相应DC亚群的表型和功能证据。使用与恒河猴交叉反应的抗人单克隆抗体，通过四色流式细胞术鉴定并分选亚群。为了动员前DC亚群，皮下注射类fms酪氨酸3激酶配体（Flt3L；100μg/kg），持续10天。在血液单核细胞的谱系-（Lin-）主要组织相容性复合体（MHC）II类+部分中鉴定出假定的前DC亚群。前单核细胞样DCs为CD11c+CD123-（白细胞介素-3Rα- [IL-3Rα-]）。前浆细胞样DCs的特征为CD11c-CD123++。Flt3L使血液中CD11c+前DC（增加7倍）和CD123++前DC亚群（增加3倍）增多。新鲜分离的CD11c+前DC亚群诱导幼稚同种异体T细胞适度增殖。在与粒细胞巨噬细胞集落刺激因子（GMCSF）和CD40L过夜培养后，两个亚群均上调表面共刺激分子，且CD11c+前DCs成为有效的同种异体刺激物。新鲜分离的CD123++前DCs表现出典型的浆细胞样形态，当与IL-3和CD40L培养72小时后，发育为成熟DC形态。在用CD40L刺激后，CD11c+前DCs分泌的IL-12p40水平增加。重要的是，单纯疱疹病毒刺激的CD123++前DCs而非CD11c+前DCs分泌干扰素-α（IFN-α）。通过流式分析和免疫组织化学在淋巴结中鉴定出相应的DC亚群，其中Flt3L给药使这两个群体均增加2至3倍。CD123+前DCs在体内病毒感染时产生IFN-α。因此，恒河猴表现出两种与人类非常相似的不同DC前体细胞群体。两者均被Flt3L动员到血液和淋巴组织中，为它们的进一步表征和可能的治疗应用提供了潜力。
