CAJ | 학술논문

目的观察脂多糖诱导的CD11b+Gr-1+髓源抑制性细胞(MDSCs)对小鼠脾脏T细胞增殖的影响,探讨其在免疫调控可能发挥的作用.方法 10只6～8周龄BALB/c小鼠随机数字表法随机分为脂多糖组和对照组各5只,分别予脂多糖或生理盐水腹腔注射；采用CD11b磁珠从脾脏组织中分选MDSCs,通过瑞氏-姬姆萨染色观察细胞形态,并用流式细胞术检测细胞表面特征分子表达情况；四唑盐(MTT)比色法测定与MDSCs在体外共培养后T细胞的增殖情况.结果脂多糖组小鼠脾脏组织中CD11b+ Gr-1+ MDSCs比例为27.4％±6.6％,较对照组(5.1％±3.8％)明显增多(t=5.06,P=0.007)；采用CD11b磁珠从注射脂多糖的小鼠脾脏中分离出的CD11b+ Gr-1+ MDSCs纯度高达84.0％±4.2％:MTT法显示在MDSCs共培养后脾脏T细胞增殖水平明显低于对照组,且此作用与MDSCs的数量呈正相关(F=46.26,P=0.000).结论 CD11b磁珠可分离提取出纯度较高的脂多糖来源的MDSCs,且脂多糖诱导的MDSCs对脾脏T细胞的增殖具有抑制作用,该作用呈剂量依赖性关系.
목적 관찰지다당유도적CD11b+Gr-1+수원억제성세포(MDSCs)대소서비장T세포증식적영향,탐토기재면역조공가능발휘적작용.방법 10지6～8주령BALB/c소서수궤수자표법수궤분위지다당조화대조조각5지,분별여지다당혹생리염수복강주사；채용CD11b자주종비장조직중분선MDSCs,통과서씨-희모살염색관찰세포형태,병용류식세포술검측세포표면특정분자표체정황；사서염(MTT)비색법측정여MDSCs재체외공배양후T세포적증식정황.결과 지다당조소서비장조직중CD11b+ Gr-1+ MDSCs비례위27.4％±6.6％,교대조조(5.1％±3.8％)명현증다(t=5.06,P=0.007)；채용CD11b자주종주사지다당적소서비장중분리출적CD11b+ Gr-1+ MDSCs순도고체84.0％±4.2％:MTT법현시재MDSCs공배양후비장T세포증식수평명현저우대조조,차차작용여MDSCs적수량정정상관(F=46.26,P=0.000).결론 CD11b자주가분리제취출순도교고적지다당래원적MDSCs,차지다당유도적MDSCs대비장T세포적증식구유억제작용,해작용정제량의뢰성관계.
Objective To explore the effects of lipopolysaccharide (LPS)-induced myeloid-derived suppressor cells (MDSCs) on the proliferation of spleen T lymphocytes.Methods BALB/c mice were randomly divided into two groups:LPS group and normal control group.They were injected intraperitoneally with LPS and normal saline solution respectively.MDSCs were separated with CD11b immunomagnetic beads from the spleen extract of mice. The morphological characteristics of MDCSs were observed by Wright-Giemsa staining and the characteristic molecules on cell surface identified by flow cytometry.And the effects of MDSCs on the in vitro proliferation of T cells were determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT).Results The proportion of MDSCs in the spleen of the LPS group was much more than that of the normal control group ( 27.4％ ± 6.6％ vs 5.1％ ± 3.8％ ; t =5.06,P =0.007 ).CD11b + Gr-1 +MDSCs could be separated by CD11b immunomagnetic beads from the spleen of mice injected with LPS at a high purity of 84.0％ +4.2％.MTT method showed that the proliferation of T cells decreased significantly after a co-cultivation with CD11 b+ M DSCs versus the control group.And it was positively correlated with the number of M DSCs ( F =46.26,P =0.000 ).Conclusions A high purity of LPS-induced myeloid-derived suppressor cells may be separated with CD11 h immunomagnetic beads.And it has dose-dependent inhibitory effects on the proliferation of the spleen T lymphocytes.