Acquired immunostimulatory phenotype of migratory CD103+ DCs promotes alloimmunity following corneal transplantation
Tomás Blanco, Hayate Nakagawa, Aytan Musayeva, Mark Krauthammer, Rohan Bir Singh, Akitomo Narimatsu, Hongyan Ge, Sara I. Shoushtari, Reza Dana
Tomás Blanco, Hayate Nakagawa, Aytan Musayeva, Mark Krauthammer, Rohan Bir Singh, Akitomo Narimatsu, Hongyan Ge, Sara I. Shoushtari, Reza Dana
View: Text | PDF
Research Article Immunology Ophthalmology

Acquired immunostimulatory phenotype of migratory CD103+ DCs promotes alloimmunity following corneal transplantation

Abstract

After transplantation, Th1-mediated immune rejection is the predominant cause of graft failure. Th1 cell sensitization occurs through complex and context-dependent interaction among antigen-presenting cell subsets, particularly CD11b+ DCs (DC2) and CD103+ DCs (DC1). This interaction necessitates further investigation in the context of transplant immunity. We used well-established preclinical models of corneal transplantation and identified distinct roles of migratory CD103+ DC1 in influencing the outcomes of the grafted tissue. In recipients with uninflamed corneal beds, migratory CD103+ DC1 demonstrate a tolerogenic phenotype that modulates the immunogenic capacity of CD11b+ DC2 primarily mediated by IL-10, suppressing alloreactive CD4+ Th1 cells via the PD-L1/PD-1 pathway and enhancing Treg-mediated tolerance via αvβ8 integrin–activated TGF-β1, thus facilitating graft survival. Conversely, in recipients with inflamed and vascularized corneal beds, IFN-γ produced by CD4+ Th1 cells induced migratory CD103+ DC1 to adopt an immunostimulatory phenotype, characterized by the downregulation of regulatory markers, including αvβ8 integrin and IL-10, and the upregulation of IL-12 and costimulatory molecules CD80/86, resulting in graft failure. The adoptive transfer of ex vivo induced tolerogenic CD103+ DC1 (iDC1) effectively inhibited Th1 polarization and preserved the tolerogenic phenotype of their physiological counterparts. Collectively, our findings underscore the essential role played by CD103+ DC1 in modulating host alloimmune responses.

Authors

Tomás Blanco, Hayate Nakagawa, Aytan Musayeva, Mark Krauthammer, Rohan Bir Singh, Akitomo Narimatsu, Hongyan Ge, Sara I. Shoushtari, Reza Dana

×

Figure 4

The effect of migratory CD103+ DC1 on corneal transplant immunity.

Options: View larger image (or click on image) Download as PowerPoint
The effect of migratory CD103+ DC1 on corneal transplant immunity. (A) B...
(A) BALB/C LR or HR recipients were transplanted as above. Allograft recipient mice were then randomized; 1 group was i.p. injected with anti–M290-SAP, while the other group was treated with anti–IgG-SAP. (B–D) Two weeks after transplantation, mice were euthanized (n = 6/group), the intracellular expression of IFN-γ by CD4+ T cells from the DLN was analyzed by flow cytometry (B), frequency (C), and expression (MFI) (D) were measured. (E) Additional mice were followed(n = 8/group) for 8 weeks to evaluate graft opacity, and Kaplan-Meier curves were plotted to evaluate graft survival. (F) Two weeks after transplantation, CD103+ DC1 were sorted by FACS from the migratory compartment of the DLN of HR recipient mice. BMDC-derived CD11b+ DC2 (iDC2) were generated with GM-CSF and stimulated with LPS. Naive CD4+CD25– T cells were sorted by MACS from the spleen of naive BALB/C mice. (G and H) Cells were cocultured for 3 days, and intracellular expression of IFN-γ by CD4+ T cells was analyzed by flow cytometry, ELISA, and MFI, and protein concentration in the supernatant was measured. (I–K) Gated CD4–CD11b+ DC2 were separated from CD103+ DC1 and analyzed by flow cytometry for MHC-II and IL-12 expression. Similar MLR was performed with iDC1 pretreated with or without IFN-γ. (L and M) CD4–CD11b+ DC2 were separated from CD103+ DC1 and analyzed by flow cytometry for MHC-II (L) and IL-12 (M) expression. (N) iDC2 and naive CD4+CD25– T cells were cocultured in the lower and iDC1 pretreated with or without IFN-γ were plated in the upper chamber of transwell system. (O and P) Gates on CD4–CD11b+ DC2 were separated from CD103+ DC1 and analyzed by flow cytometry for CD86 (O) and IL-12 (P) expression. Results are of 2 sets of triplicates with cells pooled from 3 mice on each set. Plots represent the mean(± SEM). **P < 0.01, ****P < 0.0001 (2-tailed t test [C, D, J, and K] or 1-way ANOVA [G, H, L, M, O, P] with Bonferroni correction).