Endogenous Toll-like Receptor 2 Modulates Th1/Treg-Promoting Dendritic Cells in Mice Corneal Transplantation Model

ABSTRACT

Purpose

Endogenous toll-like receptor (TLR) 2 is linked to allograft rejection in corneal transplantation. TLR2 also could modulate dendritic cell (DC) phenotype, resulting in T cell polarization. Thus, we investigated the role of endogenous TLR2 on DC development and T cell polarization during corneal rejection.

Materials and Methods

Corneas of BALB/c mice were transplanted into the eyes of C57BL/6 wild-type (WT) recipients and TLR2^−/- (KO) recipients. Graft survival and TLR2 mRNA expression were assessed. At day 14 after transplantation, to study endogenous TLR2 effects on DC development and function, surface expression of MHC classⅡ (MHCⅡ), CD86, CD80 and CD40 in ipsilateral cervical draining lymph nodes (DLNs) is measured by flow cytometry, and DC phenotype in corneas is detected by immunofluorescence. The levels of IL-12, IL-10 and IL-4 in corneas were measured by real time-qPCR (RT-qPCR). The ability of DCs to stimulate T cell polarization was assessed by IFN-γ expressions via RT-qPCR and immunohistochemistry.

Results

TLR2 mRNA expression in corneas was peaked at day 14 post-transplantation in WT group. KO group improved corneal allograft survival compared to the WT group. In addition, the KO group decreased expression of CD86, CD80 and CD40 on DCs compared to the WT group. There was no difference in MHCⅡ expression in two groups. The CD11c⁺MHCⅡ⁺CD40^high DCs could not be detected in corneas of the KO group. Moreover, the KO group decreased IL-12 (Th1-promoting cytokines) mRNA expression and increasing IL-10 (Treg-promoting cytokines) mRNA expression compared to the WT group. IL-4 (Th2-promoting cytokines) mRNA expression gained no difference between the two groups. The IFN-γ (Th1 cytokines) expression was significantly decreased in the KO group compared to the WT group.

Conclusions

Endogenous TLR2 may contribute to allogeneic corneal rejection via Th1 immunity by activating Th1-promoting DCs and suppressing Treg-promoting DCs.

KEYWORDS:

• Toll-like receptor 2
• dendritic cell
• cornea
• allograft; rejection

Disclosure

The authors reported no proprietary or commercial interest in any product mentioned or concept discussed in this article.

Additional information

Funding

This research is supported by the National Natural Science Foundation of China [Grant No. 81170887], the Natural Science Foundation of Guangdong Province [Grant No. 2017A030313602] and the Horizontal Topic Matching Funds of Nanfang Hospital, Southern Medical University [Grant No. G201202].