Immune Netw.  2011 Dec;11(6):399-405. 10.4110/in.2011.11.6.399.

Maturation-Resistant Dendritic Cells Ameliorate Experimental Autoimmune Uveoretinitis

• Oh K
• Kim YS
• Lee DS

• KMID: 2150726
• DOI: http://doi.org/10.4110/in.2011.11.6.399

Abstract

BACKGROUND
Endogenous uveitis is a chronic inflammatory eye disease of human, which frequently leads to blindness. Experimental autoimmune uveoretinitis (EAU) is an animal disease model of human endogenous uveitis and can be induced in susceptible animals by immunization with retinal antigens. EAU resembles the key immunological characteristics of human disease in that both are CD4+ T-cell mediated diseases. Dendritic cells (DCs) are specialized antigen-presenting cells that are uniquely capable of activating naive T cells. Regulation of immune responses through modulation of DCs has thus been tried extensively. Recently our group reported that donor strain-derived immature DC pretreatment successfully controlled the adverse immune response during allogeneic transplantation.
METHODS
EAU was induced by immunization with human interphotoreceptor retinoid-binding protein (IRBP) peptide(1-20). Dendritic cells were differentiated from bone marrow in the presence of recombinant GM-CSF.
RESULTS
In this study, we used paraformaldehyde-fixed bone marrow-derived DCs to maintain them in an immature state. Pretreatment with fixed immature DCs, but not fixed mature DCs, ameliorated the disease progression of EAU by inhibiting uveitogenic CD4+ T cell activation and differentiation.
CONCLUSION
Application of iBMDC prepared according to the protocol of this study would provide an important treatment modality for the autoimmune diseases and transplantation rejection.

Keyword

Dendritic cells; Maturation; Experimental Autoimmune Uveoretinitis (EAU)

MeSH Terms

Animals
Antigen-Presenting Cells
Autoimmune Diseases
Blindness
Bone Marrow
Dendritic Cells
Disease Models, Animal
Disease Progression
Eye Diseases
Eye Proteins
Graft Rejection
Humans
Immunization
Retinaldehyde
Retinol-Binding Proteins
T-Lymphocytes
Tissue Donors
Uveitis
Eye Proteins
Retinaldehyde
Retinol-Binding Proteins

Figure

• 

  Figure 1 Mature-resistant DC ameliorates EAU in mice. (A) Histology of eyes. C57BL/6 (B6) mice were intravenously injected with 5×106 BM-derived fixed immature DCs (iBMDC), fixed mature DCs (mBMDC), or media. Two days after the BMDC transfer, mice were immunized with human IRBP peptide1-20, Both eyes were removed, fixed in 4% paraformaldehyde, and embedded in paraffin. Sections (4µm) were prepared and stained with hematoxylin and eosin (original magnification, ×100). Midsagittal section for each eyes were used. V, vitreous; GL, ganglion layer; INL, inner nuclear layer; ONL, outer nuclear layer; RPE, retinal pigmented epithelium (RPE); Ch, choroid. Retina folding and infiltrations (arrows). (B) Histopathological score of EAU. Data are presented as the mean±SEM of n=10 mice for each group from two independent experiments. (C) IRBP-specific proliferation of CD4+ T cells. CD4+ T cells from draining lymph nodes were purified with magnetic beads on the 7th day after immunization, then co-cultured (2×105 cells/well) with irradiated, syngeneic splenocytes (4×105 cells/well) with 30 mM of IRBP peptide. The cultures were incubated for 3 days and pulsed with [3H]-thymidine (1 mCi/well) for the last 12 hrs. Data represent means±SD of two independent determinations with draining lymph nodes from n=3 mice/group.

• 

  Figure 2 iBMDC-pretreatment reduced the activation of CD4+ T cells. Draining lymph node cells from the mice pretreated with mBMDC, iBMDC, or media and immunized with human IRBP peptide were harvested on the 7th day after immunization. T-cell activation was analyzed using flowcytometer after staining with anti-CD4 and anti-CD62L antibodies. The data are representative of three independent determinations.

• 

  Figure 3 Intracellular cytokine expression of uveitogenic CD4+ T cells. Draining lymph node cells were harvested from the mice received mBMDC, iBMDC, or media on the 7th day after immunization with IRBP peptide and were restimulated in vitro with PMA and ionomycin as described in Materials and Methods. Cells were analyzed using flowcytometer after staining with anti-CD4 and anti-IFN-γ antibodies. The data are representative of two independent determinations.

• 

  Figure 4 Both mBMDC and iBMDC migrate into T-cell zone of the spleen and lymph nodes. B6 mice were received mBMDC and iBMDC prepared from GFP transgenic mice. On 14 days after injection with DCs, spleens and inguinal lymph nodes were frozen in OCT compound. Sections (4µm) were examined under fluorescence microscopy (original magnification, ×200). Spleen (A) and inguinal lymph node (B) from the mice received iBMDC. Spleen (C) and inguinal lymph node (D) from the mice received mBMDC. T cell zone (T) and B cell zone (B) were indicated.

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