Characterization of suppressor function of human alveolar macrophages for T lymphocyte responses to phytohemagglutinin: cellular selectivity, reversibility, and early events in T cell activation.
Characterization of suppressor function of human alveolar macrophages for T lymphocyte responses to phytohemagglutinin: cellular selectivity, reversibility, and early events in T cell activation.
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人肺泡巨噬细胞对 T 淋巴细胞对植物血凝素反应的抑制功能的表征：细胞选择性、可逆性和 T 细胞激活的早期事件。
DOI：
10.1165/ajrcmb/8.1.89
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发表时间：
1993
影响因子：
6.4
通讯作者：
Rich,EA
中科院分区：
文献类型：
--
作者：
Schauble,TL;Boom,WH;Finegan,CK;Rich,EA
材料和方法外周血单核细胞（pBMC）、单核细胞和非粘附性血液T淋巴细胞的制备通过静脉穿刺将健康非吸烟志愿者（18 - 45岁）的全血（60 - 180 ml）收集在肝素化注射器中。通过在Ficoll-Paque分离培养基（Pharmacia LKB Biotechnology，皮斯卡特维，NJ）上的密度梯度沉降分离PBMC（11）。如前所述（12），在37 ℃下，通过将PBMC粘附于血清预包被的塑料培养皿1小时来制备单核细胞。通过用塑料刮刀轻轻刮擦从培养皿中除去贴壁细胞，并以各种浓度重悬于含有青霉素50 U/ml（Squibb-Marsam，Cherry Hill，NJ）、庆大霉素5 μ g/ml（Whittaker MA Bioproducts，Walkersville，MD）和谷氨酰胺2 mM（K.C. e. Biologicals，莱内克萨，KS）。该培养基将被称为补充RPMI。通过细胞化学标准确定的赖特氏，过氧化物酶和非特异性酯酶染色的细胞离心涂片制备，粘附细胞为90至95%的单核细胞。
Materials and MethodsPreparation of Peripheral Blood Mononuclear Cells (pBMC), Monocytes, and Nonadherent Blood T LymphocytesWhole blood (60 to 180 rnl) was collected in heparinized syringes by venipuncture of healthy nonsmoking volunteers (18 to 45 yr old). PBMC were isolated by density gradient sedimentation on Ficoll-Paque separation medium (Pharmacia LKB Biotechnology, Piscataway, NJ)(11). Monocytes were prepared by adherence of PBMC to serum-precoated plastic dishes for 1 h at 37 C as described previously (12). Adherent cells were removed from dishes by gently scraping with a plastic scraper and resuspended at various concentrations in RPMI 1640 containing penicillin 50 U/rnl (Squibb-Marsam, Cherry Hill, NJ), gentamicin 5 jtg/rnl (Whittaker MA Bioproducts, Walkersville, MD), and glutamine 2 mM (K. e. Biologicals, Lenexa, KS). This medium will be referred to as supplemented RPMI. By cytochemical criteria ascertained by Wright's, peroxidase, and nonspecific esterase stains of cytospin preparations, the adherent cells were 90 to 95% monocytes.