糖基化修饰的树突状细胞疫苗激发的骨髓瘤特异性T细胞免疫反应

Glycosylation-modified dendritic cell vaccine primes the myeloma-specific T cell response

探讨经糖基化修饰的肿瘤相关糖抗原冲击树突状细胞(DC)后,所得DC疫苗对骨髓瘤患者自身T细胞的刺激作用。采用化学方法及细胞生物工程法,使骨髓瘤细胞表达新肿瘤相关抗原N-丙酰多聚唾液酸(NPrPSA);在无血清培养条件下用GM-CSF/IFN-α及TNF-α诱导培养多发性骨髓瘤(MM)患者外周血单核细胞DC,继用表达新抗原的肿瘤细胞冲击制备DC疫苗,并与MM患者自身T细胞共同温育,流式细胞仪分析CD4+CD29+、CD8+CD28+及CD69+T细胞。结果显示糖基化修饰的骨髓瘤DC疫苗与正常细胞相比,可明显诱导CD4+及CD8+T细胞的活化。糖基化修饰的DC疫苗可激发骨髓瘤特异性T细胞免疫反应,将为靶向性杀伤骨髓瘤细胞奠定基础。

To investigate the stimulatory effect of the glycosylation-modified dendritic cells vaccine pulsed with tumor-associated carbohydrate antigen （TACA） on the activation of autologous T cells of patients with myeloma, the new TACA N-propionyl polysialic acid （NPrPSA） was firstly rendered to express on myeloma cells with chemical and bio-engineering mehtods for tumor cells, and mononucleated cells from peripheral blood of patients with multiple myeloma （MM） were cultivated in serumfree culture medium containing GM-CSF/IFN-α, and TNF-α to develop dendritic cells （DC）, and then these DC were pulsed with tumor cells expressing the new TACA, thus preparing the DC vaccine. After the DC vaccine was co-cultivated with the autologous T cells of patients with MM, the CD4^＋CD29^＋ , CD8^＋ CD28^＋ and CD69^＋ T cell were analysed by cytometery. It was demonstrated that there were 6 out of 11 patients in the CD138 NPr-DC group and no one in 7 patients in the CD138^- NPr-DC group showing an increased expression of CD4CD69 （P〈0.05）; while there were 8 out of 11 patients in the CD138^＋ NPr-DC group and 1 out of 7 patients in the CD138 NPr-DC group showing an increased expression of CD8CD28. （P〈0.05） Furthermore, there were also 8 out of 11 patients in CD138 NPr-DC group and 2 out of 7 patients in the CD138 NPr-DC group showing an increased expression of CD69（P〈0.05）. These results indicate that the glycosylation-modified DC1 vaccine can activate obviously CD4^＋ and CD8^＋T cells in comparison with the normal cells, thus providing a foundation for the specific eradiation of myeloma cells.