Enhancement of specific T-lymphocyte responses by monocyte-derived dendritic cells pulsed with E2 protein of human papillomavirus 16 and human p16INK4A

Nuchsupha Sunthamala; Neeranuch Sankla; Jureeporn Chuerduangphui; Piyawut Swangphon; Wanchareeporn Boontun; Supakpong Ngaochaiyaphum; Weerayut Wongjampa; Tipaya Ekalaksananan; Chamsai Pientong

doi:10.7717/peerj.9213

Enhancement of specific T-lymphocyte responses by monocyte-derived dendritic cells pulsed with E2 protein of human papillomavirus 16 and human p16INK4A

PeerJ. 2020 May 20:8:e9213. doi: 10.7717/peerj.9213. eCollection 2020.

Authors

Nuchsupha Sunthamala^{1

2}, Neeranuch Sankla¹, Jureeporn Chuerduangphui^{2

3}, Piyawut Swangphon^{2

4}, Wanchareeporn Boontun¹, Supakpong Ngaochaiyaphum¹, Weerayut Wongjampa^{2

5}, Tipaya Ekalaksananan^{2

5}, Chamsai Pientong^{2

5}

Affiliations

¹ Department of Biology, Faculty of Science, Mahasarakham University, Maha Sarakham, Thailand.
² HPV&EBV and Carcinogenesis Research Group, Khon Kaen University, Khon Kaen, Thailand.
³ Department of Microbiology, Faculty of Science, Kasetsart University, Bangkok, Thailand.
⁴ Faculty of Medical Technology, Prince of Songkla University, Hat Yai, Songkla, Thailand.
⁵ Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.

Abstract

Introduction: Prophylactic vaccines are already available for prevention of human papillomavirus (HPV) infection. However, we still await development of therapeutic vaccines with high efficiency for stimulating specific T lymphocytes to clear HPV infection.

Objective: This study investigates the potential for subunits of human p16INK4a protein and E2 protein of HPV16 to stimulate dendritic cells and enhance the specific response of T lymphocytes against HPV-infected cells.

Methodology: Immunogenic epitopes of HPV16 E2 and p16INK4a proteins were predicted through the common HLA class I and II alleles present in the Thai population. Then, monocyte-derived dendritic cells (MDCs) were pulsed with HPV16 E2 and/or p16INK4a protein s and their maturity assessed. MDCs pulsed with either or both of these proteins at optimal concentrations were used for activation of autologous T lymphocytes and IFN-γ production was measured for specific response function.

Results: HPV16 E2 and p16INK4a proteins contain various immunogenic epitopes which can be presented by antigen-presenting cells via both HLA class I and II molecules. The stimulation of MDCs with either HPV16 E2 or p16INK4a proteins increased percentages and mean fluorescence intensity (MFI) of CD83⁺ MDCs in a dose-dependent manner. An optimum concentration of 250 ng/mL and 150 ng/mL of HPV16 E2 and p16INK4a proteins, respectively, stimulated MDCs via the MAPK pathway (confirmed by use of MAPK inhibitors). T lymphocytes could be activated by MDCs pulsed with these proteins, leading to high percentages of both CD4⁺ IFN-γ⁺ T lymphocytes and CD8⁺ IFN-γ⁺ T lymphocytes. The production of IFN-γ was higher in co-cultures containing MDCs pulsed with HPV16 E2 protein than those pulsed with p16INK4a. Interestingly, MDCs pulsed with a combination of HPV16 E2 and p16INK4a significantly increased IFN-γ production of T lymphocytes. The IFN-γ production was inhibited by both HLA class I and II blockade, particularly in co-cultures with MDCs pulsed with a combination of HPV16 E2 and p16INK4a.

Conclusions: This suggests that MDCs pulsed with both proteins enhances specific response of both CD4⁺ and CD8⁺ T lymphocytes. This study might provide a strategy for further in vivo study of stimulation of T lymphocytes for therapy of HPV-associated cancer.

Keywords: Dendritic cells; HPV16 E2; Human papillomavirus (HPV); T lymphocytes; p16INK4a.

Associated data

figshare/10.6084/m9.figshare.11493516.v1

Grants and funding

This work was performed using financial support from Khon Kean University (grant numbers 61004603 and 620008005) and scholarships under the Post-doctoral Program from Research Affairs and Graduate School, Khon Kaen University (grant no. 58222 and PD2562-12). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.