CAJ | 학술논문

目的：从黑色素瘤荷瘤小鼠脾脏中分离得到CD11b＋Gr-1＋的髓系抑制性细胞（ Myeloid derived suppressor cells ，MDSCs）。方法给C57BL/6小鼠注射LPS或者小鼠黑色素瘤细胞株B16BL6后，取小鼠的脾脏细胞，检测其中CD11b＋Gr-1＋的MDSCs 细胞的比例。通过流式细胞仪（ flow cytometry， FCM）分选出CD11b＋Gr-1＋的细胞，培养24小时后，CBA（cytometric bead array）法检测其分泌IL-10的情况；提取CD11b＋Gr-1＋细胞的总RNA，利用RT-PCR的方法检测其表达IFN-β、TGF-β、iNOS、ARG-1和IDO的情况。结果与注射LPS相比，注射B16BL6细胞株的小鼠脾脏中CD11b＋Gr-1＋的MDSCs细胞的比例更高；利用FCM分选得到CD11b＋Gr-1＋的MDSCs，进一步鉴定发现其表达IL-10、IFN-β、TGF-β、iNOS、ARG-1和IDO。与先前报道的MDSCs的特征相一致。结论可从B16BL6荷瘤小鼠脾脏中获得高纯度的MDSCs用于实验研究。
목적：종흑색소류하류소서비장중분리득도CD11b＋Gr-1＋적수계억제성세포（ Myeloid derived suppressor cells ，MDSCs）。방법급C57BL/6소서주사LPS혹자소서흑색소류세포주B16BL6후，취소서적비장세포，검측기중CD11b＋Gr-1＋적MDSCs 세포적비례。통과류식세포의（ flow cytometry， FCM）분선출CD11b＋Gr-1＋적세포，배양24소시후，CBA（cytometric bead array）법검측기분비IL-10적정황；제취CD11b＋Gr-1＋세포적총RNA，이용RT-PCR적방법검측기표체IFN-β、TGF-β、iNOS、ARG-1화IDO적정황。결과여주사LPS상비，주사B16BL6세포주적소서비장중CD11b＋Gr-1＋적MDSCs세포적비례경고；이용FCM분선득도CD11b＋Gr-1＋적MDSCs，진일보감정발현기표체IL-10、IFN-β、TGF-β、iNOS、ARG-1화IDO。여선전보도적MDSCs적특정상일치。결론가종B16BL6하류소서비장중획득고순도적MDSCs용우실험연구。
Objective To purify the CD11b+Gr-1 +MDSCs from splenic of mice bearing transplanted melanoma.Methods Splenic cells were extracted from the C57BL/6 mice which were injected with lipopolysaccharide (LPS) or B16BL6 cells and were double-stained with antibodies to CD11b and Gr-1 for flow cytometry analysis.CD11b+Gr-1 +MDSCs were sorted from C57BL/6 mice injected with B16BL6 cells, and the expression of IFN-β, TGF-β, iNOS, ARG-1, IDO and IL-10 in the purified MDSCs was checked by RT-PCR or CBA kits.Results C57BL/6 mice injected with B16BL6 cells could produce higher percentages of CD 11b+Gr-1 +MDSCs than C57BL/6 mice injected with LPS.As previously described, FCM-sorted MDSCs expressed IFN-β,TGF-β,iNOS,ARG-1,IDO and IL-10.Conclusions MDSCs can be isolated from C57BL/6 mice injected with B16BL6 cells by FCM sorting.