摘要
目的:考察Hsp70L1对肿瘤细胞免疫原性的增强作用。方法:用RT-PCR的方法,从小鼠黑色素瘤B16细胞和C57BL/6小鼠脾脏中获得TRP2153-243及Hsp70L1基因。分别插入pcDNA3.1/V5-His真核表达载体,构建pHSP70L1、pTRP和pTRP2-Hsp3种表达载体。分别转染B16肿瘤细胞并制备坏死或凋亡瘤苗,免疫C57BL/6小鼠后移植B16肿瘤细胞,观察肿瘤生长曲线,采用流式细胞术(FCM)或微量细胞毒的方法检测荷瘤小鼠细胞因子INF-γ和CTL活性。结果:将经过HSP70L1、TRP2及TRP2-HSP基因修饰的坏死或凋亡的B16肿瘤细胞免疫正常小鼠后,观察到Hsp70L1及TRP2-Hsp基因修饰的坏死瘤苗可显著抑制荷瘤鼠肿瘤的生长,并显著促进荷瘤鼠脾脏淋巴细胞CTL活性和IFN-γ产生(P<0.05,P<0.01);HSP70L1免疫刺激作用在坏死瘤苗中更明显。结论:Hsp70L1可明显提高B16瘤苗的免疫原性,且对坏死细胞瘤苗的作用更显著。
AIM:To explore the immune enhancement of Hsp70L1 in the tumor cell vaccines.METHODS:TRP2153-243 and Hsp70L1 genes were obtained by RT-PCR from B16 cells in murine melanoma and from spleens of C57BL/6 mice and then were inserted into pcDNA3.1/V5-His eukaryotic expression vectors respectively.The recombinants of pTRP2,pHSP70L1 or pTRP2-Hsp fusion gene were obtained and transfected into B16 cells respectively.TRP2153-243,HSP70L1 or TRP2-Hsp fusion gene-expressing B16 cells were then induced to necrosis by freezing-thawing or to apoptosis by mitomycin C.C57BL/6 mice were immunized with the necrotic or apoptotic B16 cells twice,then the live B16 tumor cells were transplanted into the immunized mice and the tumor growth was observed in some tumor-bearing mice.IFN-γ-producing cells in splenocytes were measured by flow cytometry and the CTL activity of spleno-lymphocyte was detected by LDH release assay.RESULTS:After the normal mice were immunized with the necrotic or apoptotic tumor vaccines modified with TRP2153-243,Hsp70L1 or TRP2-Hsp fusion genes,CTL lysis activity and IFN-γproduction from the splenic lymphocytes were promoted in the groups of Hsp70L1 and TRP2-Hsp modified tumor vaccines(P〈0.05 or P〈0.01).Additionally,the tumor growth was inhibited obviously in the groups of mice immunized with necrotic tumor vaccines(P〈0.05 or P〈0.01).However,no marked inhibition of tumor growth was observed in the groups of mice immunized with apoptotic tumor vaccines(P〈0.05).CONCLUSION:Hsp70L1 remarkably improves the immunogenicity of B16 tumor vaccines,especially that of necrotic tumor vaccines.
作者
尤红煜
张开霞
王俊霞
张焕铃
连伟光
吴为
宋淑霞
YOU Hong-yu;ZHANG Kai-xia;WANG Jun-xia;ZHANG Huan-ling;LIAN Wei-guang;WU Wei;SONG Shu-xia(Department of Molecular Biology&Key Lab.of Experimental Animal Sciences,Hebei Medical University,Shijiazhuang 050017,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2010年第4期340-343,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
河北省科技支撑资助项目(09276418D-10
052761990)
