Mycoplasma alligatoris infection promotes CD95 (FasR) expression and apoptosis of primary cardiac fibroblasts.
Mycoplasma alligatoris infection promotes CD95 (FasR) expression and apoptosis of primary cardiac fibroblasts.
复制标题
鳄鱼支原体感染促进原代心脏成纤维细胞 CD95 (FasR) 表达和凋亡。
DOI：
10.1128/cdli.12.12.1370-1377.2005
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Brown,DR
中科院分区：
文献类型：
--
作者：
Hunt,ME;Brown,DR
美洲鳄支原体引起易感宿主急性致死性原发感染。一项基因组调查表明，唾液酸酶和透明质酸酶，CD 95介导的真核细胞死亡的潜在启动子，是M.鳄鱼我们使用免疫荧光成像和流式细胞术来检测M。在体外研究了M.体内感染。通过使用针对小鼠或人CD 95的N或C末端的多克隆抗体证明了CD 95在原代培养的心脏、骨骼肌和胚胎成纤维细胞中的均匀分布。抗CD 95抗体在成纤维细胞裂解物的Western印迹上反应，具有预测的CD 95表观分子量的条带，但在对照或M的血浆中未检测到可溶性CD 95。被鳄鱼感染的鳄鱼接种M后48 h，CD 95门控心肌成纤维细胞比例增加3倍（P< 0.01）。鳄鱼感染引起心脏成纤维细胞的形态学变化，包括凋亡特征性的CD 95易位和末端脱氧核苷酸转移酶dUTP缺口末端标记凋亡分析中测量的5-溴-2 ′-脱氧尿苷（BrdU）掺入增加8倍（P< 0.16）。用抗人CD 95的激动性免疫球蛋白M激活的BrdU门控对照的比例也增加了3倍（对于肌肉，P< 0.03）。热灭活M.短吻鳄不育M.短吻鳄条件培养上清液没有影响。这是在爬行纲中的CD 95同源物的第一个报告，并建立了一个新的模型，可用于测试直接细菌与CD 95信号转导通路的上游组件的相互作用。
Mycoplasma alligatoriscauses acute lethal primary infection of susceptible hosts. A genome survey implicated sialidase and hyaluronidase, potential promoters of CD95-mediated eukaryotic cell death, as virulence factors ofM. alligatoris. We used immunofluorescence imaging and flow cytometry to examine the effects ofM. alligatorisinfection in vitro on CD95 expression and apoptosis by alligator cardiac fibroblasts, a major cell type of a target organ ofM. alligatorisinfection in vivo. A uniform distribution of CD95 in primary cultured cardiac, skeletal muscle, and embryonic fibroblasts was demonstrated by using polyclonal antibodies against the N or C terminus of mouse or human CD95. Anti-CD95 antibodies reacted on Western blots of fibroblast lysates with a band with the predicted apparent molecular weight of CD95, but soluble CD95 was not detected in plasma from control orM. alligatoris-infected alligators. The proportion of CD95-gated cardiac fibroblasts increased threefold (P< 0.01) 48 h after inoculation withM. alligatoris. Infection induced morphological changes in cardiac fibroblasts, including translocation of CD95 characteristic of apoptosis and an eightfold increase (P< 0.16) in 5-bromo-2′-deoxyuridine (BrdU) incorporation measured in a terminal deoxynucleotide transferase dUTP nick end-labeling apoptosis assay. The proportion of BrdU-gated controls activated with agonistic immunoglobulin M against human CD95 also increased threefold (P< 0.03 for muscle). Heat-inactivatedM. alligatorisand sterileM. alligatoris-conditioned culture supernatant had no effect. This is the first report of a CD95 homolog in the class Reptilia and establishes a new model that can be used to test the direct bacterial interaction with upstream components of the CD95 signal transduction pathway.