Vu Quynh, McCarthy Karin M, McCormack Joanne M, Schneeberger Eveline E
Molecular Pathology Unit, Massachusetts General Hospital East, Charlestown, MA 02129, USA.
Immunology. 2002 Apr;105(4):488-98. doi: 10.1046/j.1365-2567.2002.01382.x.
Intratracheal (IT) administration of heat-killed Listeria monocytogenes (HKL) results in an influx of macrophage and dendritic cell (DC) precursors into the lung interstitium. Low-density, FcR+, interstitial lung cells isolated from rats instilled 24 hr before with HKL or vehicle alone, were > 90% Mar1+. After culturing with granulocyte-macrophage colony-stimulating factor (GM-CSF) for 3 days, up to 24% of the loosely adherent cells were DC that stimulated allogeneic T-cell proliferation in an mixed lymphocyte reaction (MLR) assay. After only an overnight incubation with GM-CSF, however, the capacity of interstitial Mar1+ cells to stimulate HKL immune T-cell proliferation without exogenous antigen was low. By contrast, when DC were isolated as major histocompatibility complex (MHC) class II+ cells from rat lungs at 1, 3, 7 and 14 days after HKL instillation and cultured overnight with GM-CSF, their antigen presentation capacity without added exogenous antigen was robust, but declined over the 2-week period. Interestingly, hilar lymph node DC maintained their HKL antigen-presenting capacity for up to 2 weeks after instillation of HKL. Following IT administration of PKH-26 labelled HKL, fluorescent or immunolabelled organisms were detected in OX62+ DC in airway epithelium, lung interstitium and hilar lymph nodes in situ and in MHC class II+ DC isolated from these sites. We conclude that newly immigrated Mar1+ lung DC precursors, while efficient in endocytosing particulate antigens, are incapable of eliciting a significant proliferative response from HKL-sensitized T cells. By contrast, MHC class II+ DC isolated from lungs and incubated overnight with GM-CSF induce vigorous antigen-specific T-cell proliferation. Antigen-loaded lung DC in hilar lymph nodes maintain their antigen presentation capacity for up to 2 weeks.
气管内(IT)给予热灭活的单核细胞增生李斯特菌(HKL)会导致巨噬细胞和树突状细胞(DC)前体流入肺间质。从24小时前单独注入HKL或赋形剂的大鼠中分离出的低密度、FcR + 肺间质细胞,90%以上为Mar1 + 。用粒细胞巨噬细胞集落刺激因子(GM-CSF)培养3天后,高达24%的松散贴壁细胞为DC,在混合淋巴细胞反应(MLR)试验中能刺激同种异体T细胞增殖。然而,仅用GM-CSF孵育过夜后,肺间质Mar1 + 细胞在无外源性抗原的情况下刺激HKL免疫T细胞增殖的能力较低。相比之下,在HKL注入后1、3、7和14天从大鼠肺中分离出作为主要组织相容性复合体(MHC)II类 + 细胞的DC,并与GM-CSF过夜培养,它们在无添加外源性抗原的情况下的抗原呈递能力很强,但在2周内会下降。有趣的是,肺门淋巴结DC在注入HKL后长达2周内仍保持其HKL抗原呈递能力。在气管内给予PKH-26标记的HKL后,在气道上皮、肺间质和肺门淋巴结原位的OX62 + DC以及从这些部位分离出的MHC II类 + DC中检测到荧光或免疫标记的生物体。我们得出结论,新迁入的肺Mar1 + DC前体虽然在吞噬颗粒抗原方面很有效,但无法引发HKL致敏T细胞的显著增殖反应。相比之下,从肺中分离并与GM-CSF过夜孵育的MHC II类 + DC可诱导强烈的抗原特异性T细胞增殖。肺门淋巴结中负载抗原的肺DC在长达2周内保持其抗原呈递能力。