CAJ | 학술논문

目的：研究B16黑色素瘤来源的MHSP65-TCL疫苗对黑色素瘤的治疗作用，以及该疫苗在疾病治疗过程中对免疫细胞活性的影响。方法利用反复冻融的方法裂解B16黑色素瘤细胞制备肿瘤细胞裂解物（TCL），将TCL同结核分支杆菌热休克蛋白65（MHSP65）联用制备MHSP65-TCL。于不同时间点，给B16黑色素瘤荷瘤小鼠免疫MHSP65-TCL疫苗，观察该疫苗抑制黑色素细细胞瘤的情况。与此同时，为了验证MHSP65-TCL中的TCL对免疫细胞活性的影响，我们将TCL于体外作用于小鼠脾细胞，然后流式检测脾细胞CD69表达、脾细胞凋亡以及细胞培养上清液中IL-10和IFN-γ的表达情况。结果通过小鼠体内抑瘤实验发现，B16黑色素瘤来源的MHSP65-TCL疫苗在小鼠体内产生了抑制黑色素细胞瘤的作用。通过体外细胞实验发现， MHSP65-TCL中的TCL可以显著刺激体外培养的小鼠脾细胞的活化。除此之外，与这种细胞活化作用相反的，发现TCL还可以诱导一部分体外培养的小鼠脾细胞发生凋亡，并促进脾细胞分泌IL-10，同时抑制IFN-γ的分泌。结论 B16黑色素瘤来源的MHSP65-TCL疫苗对黑色素瘤具有抑制作用。这种抑制作用主要是通过活化免疫细胞产生的。MHSP65-TCL的主要成分TCL对免疫细胞还有抑制作用。这可能同TCL中存在的一些免疫细胞抑制物质有关。这些物质的明确以及去除将有助于提高MHSP65-TCL疫苗的抗肿瘤效力。
목적：연구B16흑색소류래원적MHSP65-TCL역묘대흑색소류적치료작용，이급해역묘재질병치료과정중대면역세포활성적영향。방법이용반복동융적방법렬해B16흑색소류세포제비종류세포렬해물（TCL），장TCL동결핵분지간균열휴극단백65（MHSP65）련용제비MHSP65-TCL。우불동시간점，급B16흑색소류하류소서면역MHSP65-TCL역묘，관찰해역묘억제흑색소세세포류적정황。여차동시，위료험증MHSP65-TCL중적TCL대면역세포활성적영향，아문장TCL우체외작용우소서비세포，연후류식검측비세포CD69표체、비세포조망이급세포배양상청액중IL-10화IFN-γ적표체정황。결과통과소서체내억류실험발현，B16흑색소류래원적MHSP65-TCL역묘재소서체내산생료억제흑색소세포류적작용。통과체외세포실험발현， MHSP65-TCL중적TCL가이현저자격체외배양적소서비세포적활화。제차지외，여저충세포활화작용상반적，발현TCL환가이유도일부분체외배양적소서비세포발생조망，병촉진비세포분비IL-10，동시억제IFN-γ적분비。결론 B16흑색소류래원적MHSP65-TCL역묘대흑색소류구유억제작용。저충억제작용주요시통과활화면역세포산생적。MHSP65-TCL적주요성분TCL대면역세포환유억제작용。저가능동TCL중존재적일사면역세포억제물질유관。저사물질적명학이급거제장유조우제고MHSP65-TCL역묘적항종류효력。
Objective To evaluate the therapeutic effect of MHSP65-TCL on melanoma and its effect on the activity of the immunocytes. Methods MHSP65-TCL was prepared by mixing MHSP65 with TCL derived from B16 melanoma cell lysate by repeated freezing and thawing. The MHSP65-TCL vaccine was administered in mice bearing B16 melanoma, and the changes in melanoma growth was observed. To investigate the influence of TCL in MHSP65-TCL on the activity of the immunocytes, we co-cultured TCL and mouse spleen cells in vitro, and analyzed CD69 expression on the cells, cell apoptosis, and levels of IL-10 and IFN-γin the cell culture supernatant. Results The MHSP65-TCL vaccine showed an anti-melanoma effect in the tumor-bearing mice. In the in vitro experiment, TCL in MHSP65-TCL strongly stimulated the activation of mouse spleen cells while causing apoptosis in some of the immunocytes and promoting cellular IL-10 secretion, but not IFN-γ. Conclusions MHSP65-TCL derived from B16 melanoma cells has an anti-melanoma effect mediated by the activation of immunocytes. TCL in MHSP65-TCL also has immunosuppressive effect on immunocytes possibly due to the presence of suppressive components in TCL, and identifying and eliminating these components may potentially improve the anti-tumor actovoty of MSHP65-TCL vaccine.