A novel MtHSP70-FPR1 fusion protein enhances cytotoxic T lymphocyte responses to cervical cancer cells by activating human monocyte-derived dendritic cells via the p38 MAPK signaling pathway

作者：Xiao, Meizhu; Feng, Ying; Cao, Guangming; Liu, Chongdong; Zhang, Zhenyu^*

来源：Biochemical and Biophysical Research Communications, 2018, 503(3): 2108-2116.

DOI：10.1016/j.bbrc.2018.07.167

• MtHSP70-FPR1
• Monocyte-derived dentritic cells
• Maturation
• Cytotoxic T lymphocyte
• MAPKs pathway
• Cervical cancer

Objective: To evaluate the potential effects of recombinant mycobacterium tuberculosis heat shock protein 70-formyl peptide receptor 1 (MtHSP7O-FPR1) fusion protein on human monocyte-derived dendritic cell (moDC) maturation; cytotoxic T lymphocyte (CTL) responses to cervical cancer (CC) cells; and the roles of the p38 MAPK, ERK, and JNK pathways in its transition.
Methods: Monocytes were positively selected with a MACS column with antiCD14 antibody-conjugated microbeads from umbilical cord blood. MoDCs were stimulated with MtHSP7O-FPR1, MtHSP70, a mix of MtHSP70 and FPRI, FPRI, or phosphate buffer solution (PBS) as control. Flow cytometry was used to analyze the surface molecule expression of moDCs and IFN-gamma-producing CD8(+) T cells. T cell proliferation was assessed using [3][H]-thymidine assays. The cytotoxicity of moDC-activated T cells against CC cells was evaluated by MTT assays. Cytokine production was determined by enzyme-linked immunosorbent assay. Western blotting was used to investigate protein expression.
Results: Compared with MtHSP70, MtHSP70 + FPR1, FPR1, or PBS-mediated moDCs, MtHSP7O-FPR1-pulsed moDCs expressed higher levels of CD80, CD86, CD83, HLA-DR, and CCR7; secreted more IL-12p70, TNF-alpha and IL-1 beta; and elicited stronger CTL priming and proliferation, resulting in an effective, HLA-I-dependent killing effect on CC cells. The p38 MAPK, ERK, and JNK pathways were all activated in MtHSP7O-FPRI-mediated moDC maturation, but the p38 MAPK pathway played a vital role.
Conclusions: The excellent capability of MtHSP70-FPR1 fusion protein to induce phenotypical and functional maturation of moDCs and CC-specific at. responses partly illustrates the potential clinical benefits of DC-based immunotherapy for CC.

• 出版日期2018-9-10
• 单位首都医科大学