Nicotine up-regulated 4-1BBL expression by activating Mek-PI3K pathway augments the efficacy of bone marrow-derived dendritic cell vaccination.
OBJECTIVE
To explore the role of 4-1BBL in nicotine-treated immature dendritic cells (imDCs) mediated anti-tumor effects.


METHODS
Bone marrow-derived imDCs were stimulated with nicotine and 4-1BBL expression was determinated by flow cytometry, Western blot and RT-PCR respectively. Then, the roles of 4-1BBL in nicotine-augmented DCs-dependent T cell proliferation, CTL priming and anti-tumor effects were investigated by BrdU cell proliferation assay, enzyme-linked immunospot assay and in vivo preventive effect on tumor development, respectively. Finally, using relative kinase inhibitors, the mechanism of 4-1BBL up-regulation by nicotine stimulation and the roles of Mek-PI3K signal pathways in nicotine-augmented DCs-dependent T cell proliferation were explored by Western blot and BrdU cell proliferation assay, respectively.


RESULTS
Firstly, nicotine could up-regulate 4-1BBL expression in both protein and mRNA levels. Secondly, the effects of nicotine-augmented DCs-dependent T-cell proliferation, CTL priming and anti-tumor effects could be significantly abolished by blocking CD80, CD86 and 4-1BBL activity, respectively. Thirdly, the combined blockages of CD80/CD86, CD80/4-1BBL, CD86/4-1BBL or CD80/CD86/4-1BBL signals could decrease 53.2 %, 29.6 %, 27.9 % and 54.5 % nicotine-enhanced T cell proliferation, respectively. Importantly, nicotine-induced 4-1BBL up-regulation could be decreased by the usage of Mek-PI3K pathway kinase inhibitors. The pre-treatment of Mek-p38-PI3K kinase inhibitors could obviously abolish nicotine-augmented DCs-dependent T cell proliferation.


CONCLUSIONS
CD80/CD86 and 4-1BBL are critical for nicotine augmented DCs-mediated anti-tumor effects. 4-1BBL and CD80/CD86 could be considered as potential candidates for preventive and therapeutic tumor vaccination.
Department of Immunology, Basic Medicine Science, Medical College, Xiamen University, Xiamen, 361005, Peoples Republic of China.
Hao Jie
HJ
Jin
Hua Xiu
HX
Sui
Yi Nan
YN
Wang
Feng Guang
FG
Gao
D053330:4-1BB Ligand / Q000096:biosynthesis* / Q000235:genetics; D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000627:therapeutic use; D000818:Animals; D016026:Bone Marrow Transplantation / Q000276:immunology* / Q000379:methods; D019496:Cancer Vaccines / Q000008:administration & dosage / Q000276:immunology* / Q000627:therapeutic use; D002460:Cell Line; D002478:Cells, Cultured; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000637:transplantation*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects* / Q000276:immunology; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism* / Q000502:physiology; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D009538:Nicotine / Q000494:pharmacology*; D058539:Phosphatidylinositol 3-Kinase / Q000502:physiology*; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000276:immunology
D053330:4-1BB Ligand; D000276:Adjuvants, Immunologic; D019496:Cancer Vaccines; D009538:Nicotine; D058539:Phosphatidylinositol 3-Kinase; D020930:MAP Kinase Kinase Kinases