现代肿瘤医学
現代腫瘤醫學
현대종류의학
Journal of Modern Oncology
2015年
20期
2879-2883
,共5页
蓝弘文%王于昌%徐利军%周鸿敏%李军
藍弘文%王于昌%徐利軍%週鴻敏%李軍
람홍문%왕우창%서리군%주홍민%리군
T淋巴细胞%调节性%双阴性T细胞%黑色素瘤%小鼠%近交系Babl/C%机制
T淋巴細胞%調節性%雙陰性T細胞%黑色素瘤%小鼠%近交繫Babl/C%機製
T림파세포%조절성%쌍음성T세포%흑색소류%소서%근교계Babl/C%궤제
T-lymhocytes%regulatory%double negative Tcell (DNT)%melanoma%mice%inbred Babl/C%mechanism
目的：探讨双阴性 T 细胞（DNT）促进 B16F10黑色素瘤生长的可能机制。方法：以 B16F10－luc －G5细胞系接种 Babl／C 小鼠建立种植瘤模型，IVIS 监测生长动态。流式细胞仪检测外周血中 T 淋巴细胞的比例。流式分选正常小鼠的 DNT 及 CD4＋细胞。在体内，以从正常小鼠体内获得的外源性 DNT 干预种植瘤，IVIS 监测肿瘤的生长动态，病理切片分析肿瘤组织病变。在体外，建立混合培养系统，观察 DNT 对淋巴细胞的杀伤功能及分化的影响，以及对 CD4＋细胞杀伤功能的影响。结果：荷瘤鼠 DNT 含量高于正常鼠（P ＜0．05）；CD4＋细胞含量与肿瘤大小高度负相关（R2＝－0．99）。外源性 DNT 干预后种植瘤生长速度加快（P ＜0．05），病理显示肿瘤组织中淋巴细胞浸润较对照组少。体外实验中，DNT 抑制总淋巴细胞的杀伤功能；DNT细胞直接抑制 CD4＋细胞杀伤功能。结论：DNT 促进 B16F10种植瘤生长主要机制之一可能为 DNT 直接抑制CD4＋细胞杀伤功能，从而减少了 CD4＋细胞对肿瘤的杀伤作用。
目的：探討雙陰性 T 細胞（DNT）促進 B16F10黑色素瘤生長的可能機製。方法：以 B16F10－luc －G5細胞繫接種 Babl／C 小鼠建立種植瘤模型，IVIS 鑑測生長動態。流式細胞儀檢測外週血中 T 淋巴細胞的比例。流式分選正常小鼠的 DNT 及 CD4＋細胞。在體內，以從正常小鼠體內穫得的外源性 DNT 榦預種植瘤，IVIS 鑑測腫瘤的生長動態，病理切片分析腫瘤組織病變。在體外，建立混閤培養繫統，觀察 DNT 對淋巴細胞的殺傷功能及分化的影響，以及對 CD4＋細胞殺傷功能的影響。結果：荷瘤鼠 DNT 含量高于正常鼠（P ＜0．05）；CD4＋細胞含量與腫瘤大小高度負相關（R2＝－0．99）。外源性 DNT 榦預後種植瘤生長速度加快（P ＜0．05），病理顯示腫瘤組織中淋巴細胞浸潤較對照組少。體外實驗中，DNT 抑製總淋巴細胞的殺傷功能；DNT細胞直接抑製 CD4＋細胞殺傷功能。結論：DNT 促進 B16F10種植瘤生長主要機製之一可能為 DNT 直接抑製CD4＋細胞殺傷功能，從而減少瞭 CD4＋細胞對腫瘤的殺傷作用。
목적：탐토쌍음성 T 세포（DNT）촉진 B16F10흑색소류생장적가능궤제。방법：이 B16F10－luc －G5세포계접충 Babl／C 소서건립충식류모형，IVIS 감측생장동태。류식세포의검측외주혈중 T 림파세포적비례。류식분선정상소서적 DNT 급 CD4＋세포。재체내，이종정상소서체내획득적외원성 DNT 간예충식류，IVIS 감측종류적생장동태，병리절편분석종류조직병변。재체외，건립혼합배양계통，관찰 DNT 대림파세포적살상공능급분화적영향，이급대 CD4＋세포살상공능적영향。결과：하류서 DNT 함량고우정상서（P ＜0．05）；CD4＋세포함량여종류대소고도부상관（R2＝－0．99）。외원성 DNT 간예후충식류생장속도가쾌（P ＜0．05），병리현시종류조직중림파세포침윤교대조조소。체외실험중，DNT 억제총림파세포적살상공능；DNT세포직접억제 CD4＋세포살상공능。결론：DNT 촉진 B16F10충식류생장주요궤제지일가능위 DNT 직접억제CD4＋세포살상공능，종이감소료 CD4＋세포대종류적살상작용。
Objective:To study the possible mechanism that DNT promotes the growth of B16F10 melanoma. Methods:Tumor -bearing mice were established by subcutaneous inoculation of B16F10 -luc -G5 cells.Flow cy-tometry was used to detect the composition changes of lymphocyte in peripheral blood from tumor -bearing mice and normal mice.DNT and CD4 + cells were separated by Aria Cell Sorter.DNT was injected in tumor -bearing mice and the effect on tumor growth was observed by IVIS in vivo and pathological changes.Mixture culture system (MLC)in vitro were established to reveal the mechanism of DNT negative -effect on tumor growth:DNT influence lymphocytes, killing function and differentiation,respectively were monitored by IVIS and flow cytometry after MLC.Effects of DNT direct influence on lymphoid killing function of CD4 + cells were detected by IVIS after MLC.Results:Luminescence between DNT group and tumor control group:there was no significant difference (P >0.05)of mean photon count [photon/(cm2 .ser.s)]on day 0,but significant difference (P <0.05)on day 3 ~9.Differentiation of T cells in pe-ripheral blood:In tumor control group,CD4 + increased first to peak on day 5 and then decreased,with a strong nega-tive correlation to tumor cells (bioluminescence)(R2 =-0.99).Contrast to normal mice,in tumor -bearing mice, DNT increased (P <0.05)but CD4 +CD25 +T decreased (P <0.05),there was strong correlation between two cells (R2 =-0.99).Also a strong correlation between number of CD8 + cells and DNT cells (R2 =-0.97)was also demonstrated in tumor control group.Pathology analysis of tumor:With less lymphocytes infiltration,the tumor was bigger in DNT effect group than in tumor control group.But in following day 5 ~9,the differences decreased gradual-ly.DNT influenced lymphoid killing function:)DNT influenced lymphoid killing function of total lymphocytes:24 ~72h,there was significant difference (P <0.05)of mean photon count and contents of total lymphocytes between any two groups of three.DNT also influenced lymphoid killing function of CD4 + cells.Conclusion:DNT down -regulate the immune responses and these may occur through the direct inhibition on CD4 + cells to promote the growth of mela-noma.
