CD44 and CD69 represent different types of cell-surface activation markers for human eosinophils
CD44 and CD69 represent different types of cell-surface activation markers for human eosinophils
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DOI：
10.1165/ajrcmb.18.6.3159
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发表时间：
1998-06-01
影响因子：
6.4
通讯作者：
Bochner, BS
中科院分区：
文献类型：
--
作者：
Matsumoto, K;Appiah-Pippim, J;Bochner, BS
从外周血或晚期支气管肺泡灌洗液(BAL)中提纯的嗜酸性粒细胞(EOS)与来自第五届国际人类白细胞抗原研讨会的473株单抗(MAbs)进行分析，以试图确定EOS激活的标志。采用两种策略：(1)共同寻找新鲜EOS不存在但在活体激活后(例如，在BAL液[BALF]中)或在体外与细胞因子(小于或等于10 ng/mlIL-3[IL-3]，IL-5，或粒细胞巨噬细胞集落刺激因子(GM-CSF))；(2)寻找新鲜EOS在体内激活或体外培养后表达增加的标志物。通过间接免疫荧光和流式细胞术，第一种方法显示，在测试的大约350株单抗中，只有那些识别CD69的单抗与晚期BALF EOS或细胞因子培养的EOS结合，而不与新鲜EOS结合。使用第二种方法，我们观察到在与IL-3、IL-5或GM-CSF共同培养的EOS中，CD44的表达随浓度和时间的增加而显著增加(类似于荧光强度增加2倍，P<；0.05)。但不包括干扰素-γ(高达100 ng/ml)，而其他15个成分表达的标记物的水平没有变化。尽管表达增加，但无论是新鲜的还是细胞因子培养的EOS都不能附着于CD44的配体--固定化的透明质酸。此外，同时比较来自过敏性捐赠者的低密度(比重1.085克/升)和正常密度(比重1.085克/升)EOS，一致发现CD44表达水平更高(类似于3到8倍)，但CD69在低密度EOS上没有表达。我们认为CD69和CD44代表了人类EOS不同类型的激活标志物。这些发现可能有助于评估EOS在体外和体内的激活状态。
Eosinophils (EOS) purified from peripheral blood or late-phase bronchoalveolar lavage (BAL) were analyzed with 473 monoclonal antibodies (mAbs) from the Fifth international Workshop on Human Leukocyte Antigens in an attempt to identify markers of EOS activation Two strategies were used: (1) Co look for surface markers absent on fresh EOS but present after in vivo activation (e.g., in late-phase BAL fluid [BALF]) or after in vitro culture for up to 72 h with cytokines (less than or equal to 10 ng/ml of interleukin-3 [IL-3], IL-5, or granulocyte-macrophage colony-stimulating factor [GM-CSF]); and (2) to look: for markers constitutively expressed on fresh EOS that were increased after activation in vivo or after culture in vitro. With indirect immunofluorescence and flow cytometry, the first approach revealed that among approximately 350 mAbs tested, only those recognizing CD69 became bound to late-phase BALF EOS or cytokine-cultured EOS, but not to fresh EOS. Using the second approach, we observed statistically significant concentration- and time-dependent increases in CD44 expression in EOS cultured with IL-3, IL-5, or GM-CSF (similar to 2-fold increase in fluorescence intensity, P < 0.05). but not with interferon-gamma (IFN-gamma) (up to 100 ng/ml), whereas levels of 15 other constitutively expressed markers were unchanged. Despite increased expression, neither fresh nor cytokine-cultured EOS adhered to immobilized hyaluronate, a ligand for CD44. Additionally, simultaneous comparison of hypodense (specific gravity < 1.085 g/liter) and normodense (specific gravity > 1.085 g/liter) EOS from allergic donors consistently revealed higher levels of CD44 expression (similar to 3- to 8-fold) but not CD69 expression on hypodense EOS. We conclude that CD69 and CD44 represent different types of activation markers for human EOS. These findings may be useful in assessing the state of EOS activation in vitro and in vivo.