Experimental study of the anti-melanoma effect of dentritic cells pulsed with RPL8 peptide
10.3969/j.issn.1007-3969.2014.04.007
- VernacularTitle:负载核糖体蛋白L8的树突状细胞对黑色素瘤的抑制作用
- Author:
Yan LI
;
Lingsi YIN
;
Huan YUE
;
Junqiong HUANG
;
Yonglin HU
- Publication Type:Journal Article
- Keywords:
Ribosomal protein L8;
Dendritic cells;
Melanoma;
Immunotherapy
- From:China Oncology
2014;(4):279-283
- CountryChina
- Language:Chinese
-
Abstract:
Background and purpose:Studies have shown that ribosomal protein L8 (RPL8) is shared by melanomas, gliomas and ovarian carcinomas. A peptide of RPL8 signiifcantly stimulated proliferation and cytokine expression of the hepler T cell clone and lymphocytes in melanoma patients. RPL8 may stimulate anti-tumor immunity, making RPL8 an attractive candidate for therapeutic intervention. In this study, we prepared DC pulsed by RPL8 (RPL8-DC) and investigate the anti-tumor effect of RPL8-DC on melanoma in mice.Methods: The recombinant protein was achieved through IPTG induction in E. coli and identiifed with Western blot. Bone marrow-derived DC was loaded with RPL8 protein. RPL8 and CD11c, CD80, MHC-Ⅰ, MHC-Ⅱmolecules on dentritic cells were monitored by lfuorescence microscope and FACS analysis, respectively. The anti-tumor effect of T cells in vitro was detected by MTT assay. Subcutaneous tumors were induced in C57BL/6 mice using B16 cells. The tumor volumes were measured after injection with RPL8-DC. Results:The puriifed protein was combined with speciifc antibodies. DCs pulsed by RPL8 were visualized under lfuorescent microscopy. CD11c, CD80, MHC-Ⅰ, MHC-Ⅱmolecules on DCs were up-regulated after stimulation with RPL8 and LPS. B16 cells were inhibited by T cells stimulated with RPL8-DC. The inhibition rate of tumor cells was 70%in RPL8-DC group when effector-to-target ratio was 30∶1, which was higher than PBS and DC groups. Inhibition of growth could be observed more signiifcantly in mice after the treatment with RPL8-DC. The mice receiving the therapy of RPL8-DC were able to survive much longer than the mice receiving control therapy. Conclusion:The DC pulsed by RPL8 protein can inhibit the growth of melanoma.
