Histamine release and surface CD200R1 staining as sensitive methods for assessing murine mast cell activation.
Histamine release and surface CD200R1 staining as sensitive methods for assessing murine mast cell activation.
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DOI：
10.1016/j.jim.2012.02.014
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发表时间：
2012-05-31
影响因子：
2.2
通讯作者：
Mitre, Edward
中科院分区：
文献类型：
--
作者：
Larson, David;Mitre, Edward
肥大细胞是重要的变态反应效应细胞，并参与许多其他疾病的病理过程。β-氨基己糖苷酶活性的测量是评价鼠肥大细胞活性的最常用方法，需要大量细胞，因此对于研究小鼠疾病模型中的肥大细胞的实用性有限。在这项研究中，我们评估了组胺释放的灵敏度相比，β-氨基己糖苷酶活性的肥大细胞活化的测量。在抗IgE和离子霉素刺激后，需要至少6×104个肥大细胞/ml才能检测到β-氨基己糖苷酶活性的轻微增加，而在相同的活化条件下，只需480个肥大细胞/ml即可检测到组胺释放的显著增加。这些发现表明，对于检测肥大细胞活化，组胺释放的测量远比β-氨基己糖苷酶活性的评估更敏感。此外，我们描述了一种新的流式细胞术检测小鼠肥大细胞活化的方法。当每种条件使用7.5×105个腹膜细胞并对IgE+ c-kit+细胞设门时，IgE和非IgE介导的活化后肥大细胞表面CD 200 R1的表达增加。该流式细胞术程序简单快速，刺激时间短至30分钟后表面CD 200 R1表达增加。测量组胺释放和表面CD 200 R1表达是检测小鼠肥大细胞活化的敏感方法。此外，这两种方法都可以在未纯化的腹膜细胞群上进行。由于需要的细胞数量少，这些方法是研究疾病小鼠模型中肥大细胞活化的理想方法。
Mast cells are important effector cells of allergy and are involved in the pathology of many other diseases. Measurement of β-hexosaminidase activity, the most commonly used method for evaluation of murine mast cell activity, requires a large number of cells and thus is of limited utility for studying mast cells in mouse models of disease. In this study we evaluated the sensitivity of histamine release as compared to β-hexosaminidase activity in the measurement of mast cell activation. Whereas a minimum of 6×104 mast cells per ml were required to detect slight increases in β-hexosaminidase activity after anti-IgE and ionomycin stimulation, substantial increases in histamine release could be detected under the same activating conditions with as few as 480 mast cells per ml. These findings demonstrate that measurement of histamine release is substantially more sensitive than assessment of β-hexosaminidase activity for detecting mast cell activation. Additionally, we describe a novel flow cytometric method for detecting murine mast cell activation. When using 7.5×105 peritoneal cells per condition and gating on IgE+ c-kit+ cells, mast cell expression of surface CD200R1 increased after both IgE and non IgE-mediated activation. This flow cytometric procedure was uncomplicated and rapid, with increases in surface CD200R1 expression appearing after as little as 30 minutes of stimulation time. Measuring histamine release and surface CD200R1 expression are sensitive approaches for detection of murine mast cell activation. Further, both approaches can be done on unpurified peritoneal cell populations. By requiring low numbers of cells, these approaches are ideal for investigating mast cell activation in murine models of disease.
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DOI：
10.1111/j.1365-2222.2008.03154.x
发表时间：
2009-03
期刊：
Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology
影响因子：
--
作者：
Torrero MN;Larson D;Hübner MP;Mitre E
通讯作者：
Mitre E
C-KIT配体干细胞因子可以通过对肥大细胞的影响来增强先天免疫力。
影响因子：
15.3
作者：
Maurer, M;Echtenacher, B;Hultner, L;Kollias, G;Mannel, D N;Langley, K E;Galli, S J
通讯作者：
Galli, S J
影响因子：
64.8
作者：
Malaviya, R;Ikeda, T;Abraham, SN
通讯作者：
Abraham, SN
影响因子：
4.4
作者：
Cherwinski, HM;Murphy, CA;Phillips, JH
通讯作者：
Phillips, JH
影响因子：
4.4
作者：
Torrero, Marina N.;Huebner, Marc P.;Mitre, Edward
通讯作者：
Mitre, Edward