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Archive > Volume 33 Issue 5 > 2026,33(5):486-493. DOI:10.3872/j.issn.1007-385X.2026.05.002 Prev Next
Basic Research
The specific immune response of dendritic cell vaccine pulsed with ALDH1A1 mRNA on nasopharyngeal carcinoma stem cells in vitro
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Abstract:
[Abstract] Objective: To investigate the ability of dendritic cell (DC) vaccine loaded with aldehyde dehydrogenase 1A1 (ALDH1A1) mRNA to enhance anti-nasopharyngeal carcinoma stem cell immune responses in vitro. Methods: ALDH1A1 mRNA was transcribed in vitro and electroporated into human peripheral blood-derived DCs to construct ALDH1A1-DC vaccine; EGFP mRNA was used as a transfection control. Flow cytometry was employed to analyze DC phenotypes, lymphocyte subsets, cytotoxic T lymphocyte (CTL) proliferation, and cytokine secretion. ALDH + and ALDH -C666-1 cells were sorted; expression of stem cell markers (CD24, CD133) and stemness-associated genes (OCT4, SOX2, Nanog) was analyzed. The specific cytotoxic capacity induced by ALDH1A1-DC was evaluated using the Cell Counting Kit-8 (CCK-8) assay. Results: EGFP-DC exhibited significantly higher fluorescence expression than untransfected DCs (P < 0.000 1). Moreover, ALDH1A1-DC significantly enhanced the expression of antigen-presenting molecules HLA-ABC, co-stimulatory molecules CD86, CD40, and CCR7 compared to untransfected DCs (P = 0.001 3, P = 0.005 9, P = 0.000 4, P = 0.001 9). Compared with the T lymphocyte group (T) and the DC-T group (DC-induced T lymphocytes), ALDH1A1-DC-induced T lymphocytes (ALDH1A1-DC-T) exhibited the highest proportion of CTL (P < 0.000 1), the highest proliferation index (P = 0.000 2 and P = 0.000 3, respectively), the highest proportions of CD8 +CD69 + cells (P = 0.000 5 and P < 0.000 1, respectively), and the highest proportions of CD8 + IFN-γ + subsets (P = 0.012 6 and P = 0.001 9, respectively). CCK-8 assay demonstrated that ALDH1A1-DC-T exhibited significantly enhanced cytotoxicity against ALDH + C666-1 suspension spheres compared to ALDH - spheres (P = 0.001 0). Furthermore, ALDH1A1-DC-T elicited the most potent anti-ALDH + C666-1 immune response among all lymphocyte groups tested (P = 0.001 0 and · · 486 谢云青, 等. ALDH1A1 mRNA负载树突状细胞疫苗体外诱导抗鼻咽癌干细胞免疫反应 P < 0.000 1, respectively). However, HLA-ABC blockade significantly attenuated this specific cytotoxicity (P < 0.000 1). Conclusion: mRNA transfected into DCs can be translated into protein and promote the maturation of DCs and enhance the ability of DCs to induce specific CTL immune response.
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