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目的探讨负载人肝细胞性肝癌(HCC)组织来源的CD133+肝癌细胞RNA树突状细胞(CD133+HCC RNA-DC)疫苗的免疫活性。方法采用酶消化法从人HCC组织中分离出肝癌细胞,利用流式细胞术分选出CD133+肝癌细胞,制备负载CD133+肝癌细胞RNA树突状细胞疫苗,最后用流式细胞术检测DC的表型,ELISA法测定DC分泌IL-12水平,采用混合淋巴细胞反应法检测DC在体外刺激自体淋巴细胞增殖的能力。结果 CD133+肝癌细胞RNA树突状细胞的HLA-ABC、HLA-DR、CD86、CD80、CD83表达水平分别是(96.52±2.02)%、(92.17±3.04)%、(94.25±3.28)%、(55.14±1.67)%、(40.53±2.31)%,与肝癌细胞RNA树突状细胞和成熟DC比较,差异均无统计学意义(P>0.05)。CD133+肝癌细胞RNA-DC、肝癌细胞RNA-DC、成熟DC和未成熟DC分泌IL-12的量分别为(421.50±3.12)、(418.20±1.10)、(324.20±2.19)和(102.47±4.60)pg/ml,前两者之间差异无统计学意义(P=0.14),前两者均高于后两者,差异有统计学意义(P<0.05)。CD133+肝癌细胞RNA-DC与肝癌细胞RNA-DC刺激自体T淋巴细胞增殖能力分别均强于成熟DC和无DC刺激的自体T淋巴细胞,差异均有统计学意义(P<0.05)。结论CD133+肝癌细胞RNA树突状细胞疫苗具有成熟表型,能够在体外有效刺激自体T淋巴细胞增殖。
Objective To investigate the immunological activities of CD133 + HCC RNA-DC vaccine loaded on human hepatocellular carcinoma (HCC) tissue. Methods Hepatoma cells were isolated from human HCC by enzymatic digestion, and CD133 + hepatocarcinoma cells were sorted by flow cytometry to prepare RNA dendritic cell vaccine loaded with CD133 + hepatocarcinoma cells. The DC phenotypes were detected by flow cytometry , The level of IL-12 secreted by DCs was measured by ELISA, and the ability of DCs to stimulate the proliferation of autologous lymphocytes in vitro was detected by mixed lymphocyte reaction. Results The expression levels of HLA-ABC, HLA-DR, CD86, CD80 and CD83 in dendritic cells of CD133 + hepatocarcinoma cells were (96.52 ± 2.02)%, (92.17 ± 3.04)% and (94.25 ± 3.28)%, ± 1.67)%, (40.53 ± 2.31)%, respectively. There was no significant difference between the dendritic cells and mature DCs of hepatoma cells (P> 0.05). The amounts of IL-12 secreted by CD133 + cells were (421.50 ± 3.12), (418.20 ± 1.10), (324.20 ± 2.19) and (102.47 ± 4.60), respectively pg / ml. There was no significant difference between the former two groups (P = 0.14). The former two groups were higher than the latter two groups (P <0.05). The ability of CD133 + RNA-DC and RNA-DC of liver cancer cells to stimulate autologous T lymphocyte proliferation were significantly higher than that of mature DC and DC-stimulated autologous T lymphocytes, respectively (all P <0.05). Conclusion The dendritic cell vaccine of CD133 + hepatoma cells has a mature phenotype, which can effectively stimulate the proliferation of autologous T lymphocytes in vitro.