Oligoclonal myelin-reactive T-cell infiltrates derived from multiple sclerosis lesions are enriched in Th17 cells - PubMed (original) (raw)

Oligoclonal myelin-reactive T-cell infiltrates derived from multiple sclerosis lesions are enriched in Th17 cells

Monica Montes et al. Clin Immunol. 2009 Feb.

Erratum in

Abstract

In this study, acute and chronic brain and spinal cord lesions, and normal appearing white matter (NAWM), were resected post-mortem from a patient with aggressive relapsing-remitting multiple sclerosis (MS). T-cell infiltrates from the central nervous system (CNS) lesions and NAWM were separated and characterized in-vitro. All infiltrates showed a proliferative response against multiple myelin peptides. Studies of the T-cell receptor (TCR)Vbeta and Jbeta usage revealed a very skewed repertoire with shared complementarity-determining region (CDR)3 lengths detected in all CNS lesions and NAWM. In the acute lesion, genomic profiling of the infiltrating T-cells revealed up-regulated expression of TCRalpha and beta chain, retinoic acid-related orphan nuclear hormone receptor C (RORC) transcription factor, and multiple cytokine genes that mediate Th17 cell expansion. The differentially expressed genes involved in regulation of Th17 cells represent promising targets for new therapies of relapsing-remitting MS.

PubMed Disclaimer

Conflict of interest statement

Conflict of interest: The authors do not have conflict of interest.

Figures

Figure 1.

Figure 1.

Histopathological characteristics of the AP and CP MS lesions. A) Hematoxylin and Eosin staining of the AP lesion detected perivascular inflammatory infiltrates, activated microglia and a decreased number of oligodendrocytes. B) LFB staining revealed active demyelination with phagocytosed LFB-positive myelin within macrophages (arrow). C) Axonal staining showed a relatively preserved axonal network. D), E) and F) The perivascular inflammatory infiltrate was composed of CD68+ macrophages and predominantly CD8+ lymphocytes. G) The CP lesion showed cystic changes with scattered mononuclear cells. H) Myelin staining revealed complete demyelination. I) Axonal staining showed profound axonal loss with axonal swelling and spheroids (asterisk). J), K) and L) The residual inflammatory infiltrate, which was less prominent when compared to the AP lesion, was composed of CD68+ macrophages and CD3+ lymphocytes.

Figure 2.

Figure 2.

Phenotypic characterization of lesion infiltrates. Flow cytometry plots represent CD3+ gated viable cells derived from acute and chronic lesions upon PHA expansion (10 days). CD4+ and CD8+ cell percentages of gated cells are indicated in the plot.

Figure 3.

Figure 3.

Proliferation of lesion-infiltrating T-cells against myelin peptides. The antigen specificity of NAWM and MS-lesion-derived T-cells was tested against a panel of 13 immunodominant myelin-derived peptides. The results are presented as average cpm from three wells±SD.

Figure 4.

Figure 4.

TCRVβ repertoire of infiltrating T cells derived from MS lesions and NAWM. A) Alterations of CDR3-LD are represented in a global bi-dimensional and a three-dimensional representation. The X axis displays the 26 human Vβ families, the 13 possible CDR3 lengths per Vβ family are on the Y axis, and the Vβ/HPRT transcript ratios are on the Z axis. The percentages of alterations are represented by a color code ranging from deep blue (≤ −30%) to dark red (≥ +30%) in the integrated landscapes. Black arrows indicate an example of “shared” alteration (shared CDR3 length of eight amino-acids for Vβ15 in all the lesions and NAWM). Red arrows indicate an example of lesion alteration (Vβ11 with a CDR3 length of eight amino-acids present in all lesions, but not in NAWM). B) Recurrent Vβ and CDR3 length distributions were detected in multiple infiltrates from MS lesions and NAWM. The figure presents 8 Vβ families containing shared CDR3 lengths of expanded infiltrating cells in five tested CNS regions. Shared alterations (CDR3 lengths shared by all CNS regions) are highlighted by orange boxes; shared alterations present in MS lesions, but not in NAWM are presented as pink boxes; while private alterations, corresponding to a CDR3 length that is present only in one studied lesion are presented as blue boxes.

Figure 5.

Figure 5.

Gene expression profiling of T-cell infiltrates derived from MS lesions in comparison to NAWM. Tree graphs summarize the gene expression results for IR genes with >5-fold change in MS lesions in comparison to NAWM. They represent normalized expression levels for all IR DEGs in colored bars and the clustering of genes according to their expression patterns. Superscript symbols denote DEGs in multiple lesions: *denotes DEGs shared between AP and CTh lesion, ^denotes DEGs shared between AP and CP lesion, and ~ denotes DEGs shared between CTh and CP lesion. Complete gene names are listed in the Supplementary Table 1.

Figure 6.

Figure 6.

Protein expression profiling of the T-cell infiltrate derived from the AP MS lesion. Gene array results from T-cell infiltrate derived from the acute MS lesion were compared to protein array data obtained from the supernatants of cells used for GA analysis. GA results for 17 genes were confirmed at the protein level for the genes and proteins with fold changes >1.5-fold.

Similar articles

Cited by

References

    1. Sospedra M, and Martin R, Immunology of multiple sclerosis. Annu Rev Immunol 23 (2005) 683–747. - PubMed
    1. Bielekova B, Goodwin B, Richert N, Cortese I, Kondo T, Afshar G, Gran B, Eaton J, Antel J, Frank JA, McFarland HF, and Martin R, Encephalitogenic potential of the myelin basic protein peptide (amino acids 83–99) in multiple sclerosis: results of a phase II clinical trial with an altered peptide ligand. Nat Med 6 (2000) 1167–75. - PubMed
    1. Markovic-Plese S, and McFarland HF, Immunopathogenesis of the multiple sclerosis lesion. Curr Neurol Neurosci Rep 1 (2001) 257–62. - PubMed
    1. Arenz M, Herzog-Hauff S, Meyer zum Buschenfelde KH, and Lohr HF, Antigen-independent in vitro expansion of T cells does not affect the T cell receptor V beta repertoire. J Mol Med 75 (1997) 678–86. - PubMed
    1. Muraro PA, Wandinger KP, Bielekova B, Gran B, Marques A, Utz U, McFarland HF, Jacobson S, and Martin R, Molecular tracking of antigen-specific T cell clones in neurological immune-mediated disorders. Brain 126 (2003) 20–31. - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources