Immune Netw.  2016 Aug;16(4):233-241. 10.4110/in.2016.16.4.233.

Vanilloid Receptor 1 Agonists, Capsaicin and Resiniferatoxin, Enhance MHC Class I-restricted Viral Antigen Presentation in Virus-infected Dendritic Cells

• Lee YH
• Im SA
• Kim JW
• Lee CK

• KMID: 2349884
• DOI: http://doi.org/10.4110/in.2016.16.4.233

Abstract

DCs, like the sensory neurons, express vanilloid receptor 1 (VR1). Here we demonstrate that the VR1 agonists, capsaicin (CP) and resiniferatoxin (RTX), enhance antiviral CTL responses by increasing MHC class I-restricted viral antigen presentation in dendritic cells (DCs). Bone marrow-derived DCs (BM-DCs) were infected with a recombinant vaccinia virus (VV) expressing OVA (VV-OVA), and then treated with CP or RTX. Both CP and RTX increased MHC class I-restricted presentation of virus-encoded endogenous OVA in BM-DCs. Oral administration of CP or RTX significantly increased MHC class I-restricted OVA presentation by splenic and lymph node DCs in VV-OVA-infected mice, as assessed by directly measuring OVA peptide SIINFEKL-Kb complexes on the cell surface and by performing functional assays using OVA-specific CD8 T cells. Accordingly, oral administration of CP or RTX elicited potent OVA-specific CTL activity in VV-OVA-infected mice. The results from this study demonstrate that VR1 agonists enhance anti-viral CTL responses, as well as a neuro-immune connection in anti-viral immune responses.

Keyword

Antigen presentation; Dendritic cell; MHC class I; VR1 agonist

MeSH Terms

Administration, Oral
Animals
Antigen Presentation*
Capsaicin*
Dendritic Cells*
Lymph Nodes
Mice
Ovum
Sensory Receptor Cells
T-Lymphocytes
Vaccinia virus
Capsaicin

Figure

• 

  Figure 1 VR1 agonists increase MHC class I-restricted OVA presentation in DCs infected with VV-OVA. (A) BM-DCs (1×105/well) generated from C57BL/6 (H-2b) mice were infected with VV-OVA (MOI=5), and cultured for 6 h in the presence of the indicated amounts of CP or RTX. The amounts of H-2Kb-OVA peptide complexes were assessed using CD8 OVA1.3 cells. (B, C) VR1 agonists increased MHC class I-restricted DC presentation of OVA peptides in mice infected with VV-OVA. C57BL/6 mice were orally administered CP (10 mg/kg) or RTX (0.1 mg/kg) was for 3 days, and then then were infected with VV-OVA (5×104 CFUs/mouse, i.v.). After virus infection, the mice were orally administrated the same doses of CP or RTX for consecutive 5 days. On the final day of VR1 agonist administration, DCs were isolated from the lymph nodes and spleens of the mice, and the amounts of Kb-OVA peptide complexes on the cell surface were evaluated using OVA-specific CD8 T hybridoma cells (B3Z cells). **p<0.01.

• 

  Figure 2 VR1 agonists increased the expression of SIINFEKL-H-2Kb complexes in mice infected with VV-OVA. Mice were orally administered CP or RTX for 3 days, infected with VV-OVA, and then treated with the same doses of CP or RTX for consecutive 5 days, as described in Fig. 1. DCs were isolated from the lymph nodes and spleens. (A) The amounts of Kb-OVA peptide complexes on the cell surface were directly determined using the F(ab')2 fragment of a SIINFEKL-H-2Kb-specific mAb (25-D1.16). (B) Expression levels of H-2Kb and I-Ab molecules. Blue dotted line, isotype control; green shaded area, DCs from mice not treated with a VR1 agonist (CP or RTX); red line, DCs from VR1 agonist-treated mouse.

• 

  Figure 3 VR1 agonists increased OVA-specific CTL activity in mice injected with VV-OVA. Mice were orally administered CP or RTX for 3 days, infected with VV-OVA, and then treated with the same doses of CP or RTX for consecutive 5 days, as described in Fig. 1. On the final day of VR1 agonist administration, and in vivo CTL assays were performed using CFSE-labeled syngeneic target cells. (A) Representative histograms of lymph node and spleen cells, and the percentage of specific killing of OVA[257-264] peptide-pulsed target cells in the lymph nodes and spleens are shown (n=5 for each group). Each data point represents the mean±SD of values obtained from 2 individual experiments. *p<0.05; **p<0.01.

• 

  Figure 4 CP increased the proliferation of adaptively transferred OTI cells. Mice were orally administered CP, infected with VV-OVA, and then treated with the same doses of CP for consecutive 5 days, as described in Fig. 1. On the final day of CP administration, CFSE-labeled OT-I cells were injected through the tail vein. After 4 days, proliferation of OT-I cells in the spleens and lymph nodes was determined by flow cytometry.

• 

  Figure 5 CP increased IFN-γ producing CD8 T cells in mice infected with rVV-OVA. Mice were orally administered CP, infected with VV-OVA, and then treated with the same doses of CP for consecutive 5 days, as described in Fig. 1. On the final day of CP administration, CFSE-labeled OT-I cells were injected through the tail vein. After 4 days, the lymph node and spleen cells were isolated and restimulated for 6 h with OVA[257-264] peptide-pulsed DCs. The cells were stained with an anti-CD8 mAb, then permeabilized and stained with an anti-IFN-γ mAb. (A) Representative histograms are shown. (B) The percentages of IFN-γ+ cells among total CD8+ T cells are shown graphically (n=5 for each group). **p<0.01.

• 

  Figure 6 VR1 agonists increases class I, but not class II, MHC-restricted presentation of exogenous antigen in BM-DCs. (A) BM-DCs generated from BM cells of C57BL/6 mice were treated with the indicated concentrations of CP or RTX for 4 h, and then allowed to phagocytose OVA-microspheres for 2 h. After fixing, the amounts of H-2Kb-OVA peptide complexes were assessed using CD8 OVA 1.3 cells. (B) BM-DCs generated from BM cells of BALB/c mice were treated with the indicated concentrations of CP or RTX for 4 h, and then allowed to phagocytose OVA-microspheres for 2 h. After fixing, the amounts of I-Ad-OVA peptide complexes were assessed using DOBW cells. (C, D) Effects of CP and RTX on phagocytic activity. BM-DCs generated from BM cells of C57BL/6 mice were incubated with FITC-labeled OVA-microspheres, washed, harvested, and analyzed by flow cytometry. A representative set of histograms from 3 individual experiments is shown. Shaded histograms, phagocytic activity of DCs in the absence of CP or RTX; thick line histograms, phagocytic activity of DCs in the presence of CP or RTX.

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