CAJ | 학술논문

目的：探讨小鼠沙眼衣原体肺炎感染中中性粒细胞（ polymorphonuclear neutrophils， PMN）的募集、活化及诱导其趋化的免疫学机制。方法 C57BL/6（C57）小鼠鼻腔吸入3×103 IFU（in-clusion-forming units）沙眼衣原体小鼠肺炎株（Chlamydia muridarum，Cm）建立小鼠沙眼衣原体肺炎模型。取感染后不同时间点小鼠肺组织进行组织病理学染色；通过检测肺组织内髓过氧化物酶（ myelo-peroxidase，MPO）的含量，检测 PMN 在肺组织聚集；分离小鼠肺组织单个核细胞，Giemsaˊs 染色计数肺组织炎症细胞种类；流式细胞术检测 CD11b＋ Gr1＋ PMN 细胞百分率及 CD11b＋ PMN 活性水平；应用RT-PCR 技术检测 Cm 感染后肺组织内与 PMN 募集相关的趋化性细胞因子（MIP-2、LIX、KC、MCP-1）的表达，探讨小鼠 Cm 呼吸道感染过程中 PMN 趋化机制。结果一定剂量的 Cm 呼吸道吸入诱导C57小鼠衣原体肺炎，与对照组（感染第0天）比较，肺组织内有大量炎性细胞浸润，主要包括 PMN、单核细胞、淋巴细胞，其中 PMN 在肺内的聚集早于单核细胞；Cm 感染诱导小鼠肺组织 MPO 含量升高，在感染第7天达到高峰，第14天虽然有所下降，但仍明显高于对照组；流式结果显示，Cm 感染后， CD11b＋Gr1＋细胞百分比、PMN 总数均显著高于对照组，同时 PMN CD11b 分子的表达在感染后也显著升高，说明 Cm 感染能诱导小鼠肺组织内 PMN 大量聚集并活化。RT-PCR 结果显示，Cm 感染后小鼠肺组织内与 PMN 募集相关的趋化性细胞因子（MIP-2、LIX、KC、MCP-1）的含量明显升高，提示 Cm 感染上调 PMN 相关趋化性细胞因子的表达，从而吸引大量 PMN 在肺部聚集参与炎症反应。结论一定剂量的 Cm 可以诱导感染小鼠肺组织内 PMN 相关趋化性细胞因子的分泌，从而使 PMN 在肺组织内大量浸润并活化，在抵抗衣原体感染中发挥重要作用。目的：探討小鼠沙眼衣原體肺炎感染中中性粒細胞（ polymorphonuclear neutrophils， PMN）的募集、活化及誘導其趨化的免疫學機製。方法 C57BL/6（C57）小鼠鼻腔吸入3×103 IFU（in-clusion-forming units）沙眼衣原體小鼠肺炎株（Chlamydia muridarum，Cm）建立小鼠沙眼衣原體肺炎模型。取感染後不同時間點小鼠肺組織進行組織病理學染色；通過檢測肺組織內髓過氧化物酶（ myelo-peroxidase，MPO）的含量，檢測 PMN 在肺組織聚集；分離小鼠肺組織單箇覈細胞，Giemsaˊs 染色計數肺組織炎癥細胞種類；流式細胞術檢測 CD11b＋ Gr1＋ PMN 細胞百分率及 CD11b＋ PMN 活性水平；應用RT-PCR 技術檢測 Cm 感染後肺組織內與 PMN 募集相關的趨化性細胞因子（MIP-2、LIX、KC、MCP-1）的錶達，探討小鼠 Cm 呼吸道感染過程中 PMN 趨化機製。結果一定劑量的 Cm 呼吸道吸入誘導C57小鼠衣原體肺炎，與對照組（感染第0天）比較，肺組織內有大量炎性細胞浸潤，主要包括 PMN、單覈細胞、淋巴細胞，其中 PMN 在肺內的聚集早于單覈細胞；Cm 感染誘導小鼠肺組織 MPO 含量升高，在感染第7天達到高峰，第14天雖然有所下降，但仍明顯高于對照組；流式結果顯示，Cm 感染後， CD11b＋Gr1＋細胞百分比、PMN 總數均顯著高于對照組，同時 PMN CD11b 分子的錶達在感染後也顯著升高，說明 Cm 感染能誘導小鼠肺組織內 PMN 大量聚集併活化。RT-PCR 結果顯示，Cm 感染後小鼠肺組織內與 PMN 募集相關的趨化性細胞因子（MIP-2、LIX、KC、MCP-1）的含量明顯升高，提示 Cm 感染上調 PMN 相關趨化性細胞因子的錶達，從而吸引大量 PMN 在肺部聚集參與炎癥反應。結論一定劑量的 Cm 可以誘導感染小鼠肺組織內 PMN 相關趨化性細胞因子的分泌，從而使 PMN 在肺組織內大量浸潤併活化，在牴抗衣原體感染中髮揮重要作用。
목적：탐토소서사안의원체폐염감염중중성립세포（ polymorphonuclear neutrophils， PMN）적모집、활화급유도기추화적면역학궤제。방법 C57BL/6（C57）소서비강흡입3×103 IFU（in-clusion-forming units）사안의원체소서폐염주（Chlamydia muridarum，Cm）건립소서사안의원체폐염모형。취감염후불동시간점소서폐조직진행조직병이학염색；통과검측폐조직내수과양화물매（ myelo-peroxidase，MPO）적함량，검측 PMN 재폐조직취집；분리소서폐조직단개핵세포，Giemsaˊs 염색계수폐조직염증세포충류；류식세포술검측 CD11b＋ Gr1＋ PMN 세포백분솔급 CD11b＋ PMN 활성수평；응용RT-PCR 기술검측 Cm 감염후폐조직내여 PMN 모집상관적추화성세포인자（MIP-2、LIX、KC、MCP-1）적표체，탐토소서 Cm 호흡도감염과정중 PMN 추화궤제。결과일정제량적 Cm 호흡도흡입유도C57소서의원체폐염，여대조조（감염제0천）비교，폐조직내유대량염성세포침윤，주요포괄 PMN、단핵세포、림파세포，기중 PMN 재폐내적취집조우단핵세포；Cm 감염유도소서폐조직 MPO 함량승고，재감염제7천체도고봉，제14천수연유소하강，단잉명현고우대조조；류식결과현시，Cm 감염후， CD11b＋Gr1＋세포백분비、PMN 총수균현저고우대조조，동시 PMN CD11b 분자적표체재감염후야현저승고，설명 Cm 감염능유도소서폐조직내 PMN 대량취집병활화。RT-PCR 결과현시，Cm 감염후소서폐조직내여 PMN 모집상관적추화성세포인자（MIP-2、LIX、KC、MCP-1）적함량명현승고，제시 Cm 감염상조 PMN 상관추화성세포인자적표체，종이흡인대량 PMN 재폐부취집삼여염증반응。결론일정제량적 Cm 가이유도감염소서폐조직내 PMN 상관추화성세포인자적분비，종이사 PMN 재폐조직내대량침윤병활화，재저항의원체감염중발휘중요작용。
Objective To investigate the possible mechanism of aggregation and activation of neu-trophils(polymorphonuclear neutrophils,PMN)in mice with chlamydial pneumonitis. Methods C57BL/ 6 mice were inoculated intranasally with 3×103 inclusion-forming units(IFU)of Chlamydia muridarum(Cm) to induce the murine model of chlamydial pneumonitis. Samples of lung tissues collected at different time points after infection were stained by hematoxylin and eosin for histopathological assessment of inflammation. The levels of myelo-peroxidase(MPO)were detected for the evaluation of PMN aggregation. The mononu-clear cells were isolated from lung tissues. The inflammatory cells were counted with Giemsaˊs staining. CD11b+Gr1+ cell population and CD11b expression in lung mononuclear cells were analyzed by flow cytome-try. The expression of chemokines(MIP-2,LIX,KC and MCP-1)in lung tissues at mRNA level was meas-ured by RT-PCR. Results Chlamydial pneumonitis was induced in mice by intranasal inoculation of 3×103 IFU of Cm. Compared with the mice from control group,large amounts of inflammatory cells including PMN, monocytes and lymphocytes were induced in lung tissues of mice with Cm infection. PMN responded earlier than monocytes to the infection. The levels of MPO were significantly increased in mice with Cm infection and reached the highest level on the 7th day after infection. A decline in MPO levels was observed on the 14th day but the levels were still higher than those on day 0. The percentages and total numbers of CD11b+Gr1+ cells were significantly increased after Cm infection. Moreover,an increased expression of PMN CD11b was also detected by flow cytometry. The expression of chemokines(MIP-2,LIX,KC and MCP-1)was in-creased in lung tissues of mice after Cm infection. The results of the study indicated that Cm infection in-duced the expression of PMN chemoattractants,resulting in the recruitment of PMN. Conclusion The infil-tration and activation of PMN in lung tissues of mice were induced by Cm infection through increasing the ex-pression of chemokines. PMN played an important role in immune responses against Cm infection.