Immunological Competence of Dendritic Cell Vaccine Loaded with CD133+ Hepatocellular Carcinoma Cell RNA
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摘要: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。
Abstract:ObjectiveTo investigate the immunological competence of dendritic cell loaded with CD133+ hepatocellular carcinoma cell RNA (CD133+ HCC RNA-DC) vaccine.
MethodsHCC cells were separated from hepatocellular carcinoma tissues through Enzyme Digestion, and then CD133+ HCC cells were sorted by flow cytometry. CD133+ HCC RNA-DC vaccine was obtained. Flow cytometry was used to detect the phenotype of DC, and ELISA was applied to determine the level of DC's secretion of IL-12. Mixed lymphocyte reaction was applied to test the ability of DC to stimulate the proliferation of autologous T lymphocytes in vitro.
ResultsThe expression levels of HLA-ABC, HLA-DR, CD86, CD80 and CD83 in CD133+ HCC RNA-DC group were (96.52±2.02)%, (92.17±3.04)%, (94.25±3.28)%, (55.14±1.67)% and (40.53±2.31)%, respectively. Phenotypes expression levels were not significantly different among CD133+ HCC RNA-DC, HCC-DC and mature DC. The secretion of IL-12 in CD133+ HCC RNA-DC group, HCC-DC group, mature DC group and immature DC group were (421.50±3.12), (418.20±1.10), (324.20±2.19) and (102.47±4.60) pg/ml, respectively; the difference of the former two had no statistical significance (P=0.14), and the amounts of the former two were higher than that of the latter two (P < 0.05). The proliferation of autologous T lymphocytes stimulated by CD133+ HCC RNA-DC and HCC-DC were stronger than those by mature DC and no DC (P < 0.05).
ConclusionThe dendritic cell vaccine loaded with CD133+ hepatocellular carcinoma cell RNA (CD133+ HCC RNA-DC) has mature phenotype and can effectively stimulate the proliferation of autologous T lymphocytes in vitro.
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Key words:
- CD133 /
- Hepatocellular carcinoma /
- RNA /
- Dendritic cell vaccine
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图 1 CD133+肝癌细胞RNA-DC形态图
Figure 1 Morphology of dendritic cell loaded with CD133+ hepatocellular carcinoma cell RNA (CD133+ HCC RNA-DC)
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图 2 负载CD133+肝癌细胞RNA的DC抗原递呈相关分子表达情况
Figure 2 Phenotypes expression of CD133+ HCC RNA-DC
表 1 不同DC的抗原递呈相关分子表达情况 (x±s, %)
Table 1 Phenotypes expression of CD133+ HCC RNA-DCs, HCC-DCs and mature DCs (x±s, %)
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