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目的:研究乳腺癌患者腋下淋巴结单个核细胞来源的树突状细胞(DC)诱导抗原特异性细胞毒T淋巴细胞(CTL)的能力。方法:以多种细胞因子联合诱导腋下淋巴结单个核细胞中的贴壁细胞为DC,非贴壁细胞与IL-2共培养后诱导成为肿瘤区域引流淋巴结细胞(TDLNC),用自体乳腺癌细胞冻融抗原刺激DC,并和TDLNC共培养,以诱导肿瘤抗原特异性CTL。结果:淋巴结细胞经体外培养后,贴壁细胞在DC诱导前,其特异性表面标志物CD1a、CD83、CD86的百分比(%)分别为11.0±2.4、26.6±5.2和33.0±6.1,经与rhGM-CSF、rhIL-4共同培养,并经自体肿瘤冻融抗原加TNF-α诱导后3种标志物的水平升高,其百分比(%)分别为50.2±5.7、60.5±16.5和56.2±16.4(P<0.01)。TDLNC中CD3+和CD8+细胞含量(%)分别为73.9±2.2和32.8±3.2;经DC和肿瘤抗原诱导后,DC-Ag-TDLNC中的CD3+和CD8+细胞含量升高,其百分比(%)分别为82.7±2.8和62.5±2.5(P<0.01)。结论:乳腺癌患者腋下引流淋巴结中的单个核细胞在rhGM-CSF和rhIL-4刺激活化和自体肿瘤抗原及TNF-α的诱导下,可以分化为典型的DC。成熟的DC具有较强的抗原提呈功能,可以促进TDLNC增殖、分化为抗原特异性CTL。
Objective: To investigate the ability of dendritic cells (DCs) derived from axillary lymph node cells of breast cancer to induce antigen-specific cytotoxic T lymphocytes (CTLs) in breast cancer patients. Methods: The adherent cells in axillary lymph node mononuclear cells were induced by a combination of cytokines. The non-adherent cells were co-cultured with IL-2 and then induced to become the draining lymph node cells (TDLNC) in the tumor area. Autologous breast cancer cells Frozen-thawed antigen stimulated DCs and co-cultured with TDLNC to induce tumor antigen-specific CTLs. Results: The percentage of specific surface markers CD1a, CD83, and CD86 of adherent cells after cultured in vitro was 11.0 ± 2.4,26.6 ± 5.2 and 33.0 ± 6.1, respectively. After treated with rhGM -CSF and rhIL-4 were co-cultured and the levels of three markers increased after induction of TNF-α by autologous tumor freeze-thaw and the percentages (%) were 50.2 ± 5.7, 60.5 ± 16.5 and 56.2 ± 16.4 P <0.01). The percentage (%) of CD3 + and CD8 + cells in TDLNC were 73.9 ± 2.2 and 32.8 ± 3.2, respectively. After induced by DC and tumor antigen, the percentage of CD3 + and CD8 + cells in DC-Ag- 82.7 ± 2.8 and 62.5 ± 2.5 (P <0.01). Conclusion: The mononuclear cells in axillary lymph node of breast cancer patients can differentiate into typical DCs under the stimulation of rhGM-CSF and rhIL-4 stimulation and autologous tumor antigen and TNF-α. Mature DC has a strong antigen-presenting function, can promote TDLNC proliferation, differentiation into antigen-specific CTL.