Résumé
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.
Résumé
The complete nucleotide sequence of a cDNA clone encoding weever ribosomal protein L8, WeevL8, is described here. The WeevL8 cDNA has 848 nucleotides in full length and encodes a 257 amino-acid protein with a calculated molecular mass of 28.02 kDa. Two conserved domains have been identified in WEEVL8. It was concluded fromsequence alignment that WeevL8 gene was quite conservative, consistent with its role as a house-keeping gene. A phylogenetic analysis made by L8 protein showed the similar phylogenetic relationship to that with 18s rDNA. The high similarity supports the notion that ribosomal protein L8 can also be used as phylogenetic criterion.
Résumé
The complete nucleotide sequence of a cDNA clone encoding weever ribosomal protein L8, WeevL8, is described here. The WeevL8 cDNA has 848 nucleotides in full length and encodes a 257 amino-acid protein with a calculated molecular mass of 28.02 kDa. Two conserved domains have been identified in WEEVL8. It was concluded fromsequence alignment that WeevL8 gene was quite conservative, consistent with its role as a house-keeping gene. A phylogenetic analysis made by L8 protein showed the similar phylogenetic relationship to that with 18s rDNA. The high similarity supports the notion that ribosomal protein L8 can also be used as phylogenetic criterion.
Résumé
The complete nucleotide sequence of a cDNA clone encoding weever ribosomal protein L8, WeevL8, is described here. The WeevL8 cDNA has 848 nucleotides in full length and encodes a 257 amino-acid protein with a calculated molecular mass of 28.02 kDa. Two conserved domains have been identified in WEEVL8. It was concluded fromsequence alignment that WeevL8 gene was quite conservative, consistent with its role as a house-keeping gene. A phylogenetic analysis made by L8 protein showed the similar phylogenetic relationship to that with 18s rDNA. The high similarity supports the notion that ribosomal protein L8 can also be used as phylogenetic criterion.