Activation of antigen-presenting cells by microbial products breaks self tolerance and induces autoimmune disease - PubMed (original) (raw)

Activation of antigen-presenting cells by microbial products breaks self tolerance and induces autoimmune disease

Hanspeter Waldner et al. J Clin Invest. 2004 Apr.

Abstract

We describe the generation of mice that express a transgenic T cell receptor (TCR) (5B6) specific for the encephalitogenic myelin proteolipid protein (PLP) peptide 139-151, on the experimental autoimmune encephalomyelitis-resistant (EAE-resistant) B10.S background. Despite harboring a high frequency of self-reactive T cells, 5B6 transgenic mice on the B10.S background rarely develop spontaneous EAE, which is in striking contrast to 5B6 transgenic mice on the EAE-susceptible SJL background. The relative resistance to spontaneous EAE in transgenic B10.S mice is not due to deletion or anergy of T cells, but appears to be controlled by APCs. Analysis of APCs revealed a lower activation state and a lower T cell-activating capacity for APCs from B10.S mice than for those from EAE-susceptible SJL mice. When APCs in 5B6 transgenic B10.S mice were activated, for example, via TLR9 or TLR4, T cell tolerance was broken, resulting in EAE. Our findings demonstrate that activation of APCs via innate immune receptors can break self tolerance and trigger the development of autoimmunity even in a genetically resistant strain. These findings suggest that the development of autoimmune diseases such as multiple sclerosis is determined at least partly by the endogenous activation state of APCs.

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Figures

Figure 1

Figure 1

Flow cytometry analysis of thymocytes and peripheral T cells in 5B6 transgenic B10.S mice. (A and B) T cells from thymi (A) and spleens (B) of mice were stained with the indicated antibodies (PE- or allophycocyanin-conjugated anti-CD4, FITC-conjugated anti-CD8, and PE-conjugated anti–TCR Vβ6). Dot plots representing two-color flow cytometry analysis of 5B6 transgenic mice with or without RAG-2 deficiency (TG/RAG-2–/– or TG, respectively) and nontransgenic littermates (NLM) are shown. Numbers in quadrants refer to percentages of gated cell populations. Total numbers of spleen cells are shown above dot plots.

Figure 2

Figure 2

Responses of 5B6 transgenic T cells to PLP139–151. (A) Proliferative response of 5B6 transgenic T cells to PLP139–151 presented by DAS cells. CD4-enriched T cell samples from 5B6 transgenic SJL and B10.S mice were stimulated with the indicated numbers of PLP139–151–pulsed DAS cells. T cell proliferation was assessed by [3H]thymidine incorporation assay. The mean cpm ± SD of triplicate cultures are shown. (B) Proliferative response of T cells from 5B6 transgenic B10.S or SJL mice to PLP139–151 presented by syngeneic APCs. Splenocyte samples from unimmunized 5B6 transgenic and nontransgenic littermate (NLM) B10.S or SJL mice were enriched for CD4+ T cells and were cultured with irradiated splenocytes from nontransgenic littermates in the presence of the indicated concentrations of PLP139–151 or control peptide PLP178–191. Proliferative responses were determined by [3H]thymidine incorporation assay. The mean cpm ± SD of triplicate cultures of one experiment representative of three are shown. (C) IL-2 response to PLP139–151 of T cells from 5B6 transgenic B10.S or SJL mice. Supernatants from cultures in B were assayed in duplicate by ELISA for cytokine production. Representative data from one of three experiments are shown. (D) INF-γ response of spleen cells from 5B6 transgenic B10.S or SJL mice to PLP139–151 stimulation. Culture supernatants from 5B6 transgenic SJL or B10.S whole spleen cells stimulated with the indicated concentrations of PLP139–151 were assayed in duplicate by ELISA for INF-γ production.

Figure 3

Figure 3

Activation/maturation state and T cell-activating capacity of APCs. (A–E) Spleen cells (A–C) or BM-derived APCs (D and E) from wild-type B10.S and SJL mice were stained with PE-conjugated antibodies against the indicated subsets of APCs and biotinylated anti–I-Aq/s. The level of MHC class II expression was determined on gated APC subsets as indicated (%) (A and D) by flow cytometry and is shown as mean of fluorescence intensity (MFI) (B, C, and E). Data from one of three experiments with similar data are shown. (F) The indicated numbers of irradiated spleen cells from wild-type SJL or B10.S mice as APCs in presence of PLP139–151 were incubated with 5B6 transgenic SJL T cells and were subsequently pulsed with [3H]thymidine. T cell proliferation was measured by [3H]thymidine incorporation assay. Values are shown as mean cpm ± SD of triplicate wells. One experiment representative of three is shown.

Figure 4

Figure 4

CpG ODN induces expansion of APC subsets and upregulation of MHC class II and CD86. Spleen cells from B10.S mice were stimulated with CpG ODN or non–CpG ODN. Spleen cells were subsequently stained with FITC-conjugated anti-CD11b, anti-CD11c, and anti-CD19 and PE-conjugated anti-CD86 or biotinylated anti–I-Aq/s. Expression of CD86 and MHC class II was determined on CD11c+-, CD11b+-, or CD19+-gated cells. Dotted lines indicate isotype controls. Numbers refer to mean of fluorescence intensity of specific staining (upper number) shown on gated APC subsets (%).

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