Analysis of Autoreactive CD4 T Cells in Experimental Autoimmune Encephalomyelitis after Primary and Secondary Challenge Using MHC Class II Tetramers

Bischof, Felix; Hofmann, Matthias; Schumacher, Ton N.; Vyth‐Dreese, Florry A.; Weissert, Robert; Schild, Hansjörg; Kruisbeek, Ada M.; Melms, Arthur

doi:10.4049/jimmunol.172.5.2878

Cited by 43 publications

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“…It has long been recognized that the development of autoimmunity in the CNS is preceded by activation of self-specific lymphocytes in peripheral lymphoid organs (5,6), but the characteristics of the peripheral immune response that determine the development CNS autoimmunity have not yet been resolved.…”

Section: Discussionmentioning

confidence: 99%

“…Recombinant MHC class II tetramers were produced essentially as described (6). The proteins were expressed in suspension cultures of Sf9 cells in protein-free insect medium (BD Pharmingen).…”

Section: Production Of Mhc II Tetramersmentioning

confidence: 99%

“…Single-cell suspensions were prepared from draining lymph nodes (DLN) and spleen as described (6). Cells were incubated with 20 g/ml tetramer in 20 l of PBS containing 1% serum albumin and 0.02% NaN 3 at 4°C overnight, stained on ice with additional Abs for 20 min and directly before analysis with 1 g/ml propidium iodide.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…Although PT is not an absolute requirement for EAE induction, it greatly facilitates the development of the disease. Mice immunized with PLP 139 -151 / CFA which received in addition PT on the day of immunization developed severe clinical disease with a relapsing remitting disease course ( In PLP 139 -151 -induced EAE in SJL mice, autoreactive T lymphocytes start to infiltrate the CNS at day 8 after immunization (6). To assess whether the disease-enhancing effect of PT is due to its effects on the peripheral activation of the immune response or by alterations at the blood-brain barrier, PLP 139 -151 /CFA immunized mice received PT on day 6 after immunization, 2 days before entry of self-reactive lymphocytes into the CNS.…”

Section: Pt Induces Eae In Neuroantigen/cfa Immunized Mice When Givenmentioning

confidence: 99%

“…CD4 ϩ helper T cells specific for Ags in the CNS play a central role in orchestrating the effector processes of this self-directed immune response (3,4). Autoimmune CNS inflammation in MS and EAE is preceded by activation of CD4 ϩ T cells specific for CNS Ags in peripheral lymphoid organs (5,6), but peripheral activation of CNS-specific T cells is not necessarily sufficient to induce autoimmunity in the CNS. In this study, we assessed the prerequisites of a peripheral immune response that lead to the development of CNS autoimmunity.…”

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confidence: 99%

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Induction of Autoimmunity by Expansion of Autoreactive CD4+CD62Llow Cells In Vivo

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et al. 2006

The Journal of Immunology

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The prerequisites of peripheral activation of self-specific CD4+ T cells that determine the development of autoimmunity are incompletely understood. SJL mice immunized with myelin proteolipid protein (PLP) 139–151 developed experimental autoimmune encephalomyelitis (EAE) when pertussis toxin (PT) was injected at the time of immunization but not when injected 6 days later, indicating that PT-induced alterations of the peripheral immune response lead to the development of autoimmunity. Further analysis using IAs/PLP139–151 tetramers revealed that PT did not change effector T cell activation or regulatory T cell numbers but enhanced IFN-γ production by self-specific CD4+ T cells. In addition, PT promoted the generation of CD4+CD62Llow effector T cells in vivo. Upon adoptive transfer, these cells were more potent than CD4+CD62Lhigh cells in inducing autoimmunity in recipient mice. The generation of this population was paralleled by higher expression of the costimulatory molecules CD80, CD86, and B7-DC, but not B7-RP, PD-1, and B7-H1 on CD11c+CD4+ dendritic cells whereas CD11c+CD8α+ dendritic cells were not altered. Collectively, these data demonstrate the induction of autoimmunity by specific in vivo expansion of CD4+CD62Llow cells and indicate that CD4+CD62Llow effector T cells and CD11c+CD4+ dendritic cells may be attractive targets for immune interventions to treat autoimmune diseases.

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Section: Discussionmentioning

confidence: 99%

Section: Production Of Mhc II Tetramersmentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

Section: Pt Induces Eae In Neuroantigen/cfa Immunized Mice When Givenmentioning

confidence: 99%

mentioning

confidence: 99%

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Induction of Autoimmunity by Expansion of Autoreactive CD4+CD62Llow Cells In Vivo

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Doster

Lange

et al. 2006

The Journal of Immunology

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Detection of low‐frequency human antigen‐specific CD4⁺ T cells using MHC class II multimer bead sorting and immunoscope analysis

et al. 2004

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MHC class II tetramers are attractive tools to study antigen-specific CD4 + T cell responses in various clinical situations in humans. HLA-DRA1*0101/DRB1*0401 MHC class II heterodimers were produced as empty molecules using the Drosophila melanogaster expression system. Peptide binding experiments revealed that these molecules could be loaded efficiently with appropriate MHC class II tumor epitopes. Interestingly, MHC class II tetramer staining was influenced by modifications in membrane lipid rafts, and could in itself induce activation changes of stained CD4 + T cells at 37 C. In order to increase the threshold of detection of poorly represented peripheral antigen-specific CD4 + T cells, we combined cell sorting using MHC class II multimer beads together with TCR analysis using the immunoscope technology. This strategy greatly increased the sensitivity of detection of specific CD4 + T cells to frequencies as low as 4Â10 -6 among peripheral blood mononuclear cells. Such a combined approach may have promising applications in the immunomonitoring of patients under vaccination protocols to tightly follow induced antigen-specific CD4 + T cells expressing previously identified TCR.

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A new T‐cell activation mode for suboptimal doses of antigen under the full activation of T cells with different specificity

et al. 2015

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Loss of tolerance for autoantigens is a common feature in autoimmune diseases. Bystander T-cell activation is the activation of T cells to produce functional changes through TCR-independent stimulation. Although bystander activation may be related to tolerance loss to multiple autoantigens, the activation mechanism of T cells directed to an autoantigen with limited amount is not clear. We investigated an activation mode of T cells (designated as "associator T cells") directed to a suboptimal dose of cognate antigen X in the presence of fully activated T cells (designated as "responder T cells") directed toan optimal dose of antigen Y. In in vitro coculture, the activation of associator T cells was dependent on the presentation of antigen X, and soluble factors from activated responder T cells were not sufficient. Therefore, we conclude this activation mode is different from bystander activation and named it "extended antigen priming (EAP)". T cells with EAP showed a different phenotype compared to conventionally primed cells, suggesting the unique nature of EAP. Intriguingly, EAP was dependent on the CD40-CD40L signaling pathway. Thus, the EAP model is a T-cell activation mode for suboptimal dose of antigen and presumably related to the immune response to autoantigens in autoimmune status.Keywords: Antigen specificity · Autoimmunity · Autoreactive T cells · Bystander activation · Epitope spreading Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionThe immune system is a network to protect the host from attacks by pathogens. In contrast, immune activation to self-antigens is suppressed by self-tolerance. However, in autoimmunity, selftolerance is broken in certain sets of autoantigens at both T-cell and B-cell levels. In the patients' sera of systemic lupus erythematosus, one of the prototypical systemic autoimmune diseases, Correspondence: Dr. Keishi Fujio e-mail: kfujio-tky@umin.ac.jp more than 100 types of autoantibodies were described [1]. Several types of autoantibodies such as anti-nuclear, anti-ds DNA, anti-phospholipid, anti-Ro, anti-La, anti-ribonucleoprotein, and anti-Sm are detected not only at the time of diagnosis but also before diagnosis of SLE for more than 5 years [2]. In patients with rheumatoid arthritis (RA), a systemic inflammatory disorder that principally affects synovial joints, autoantibodies recognizing the Fc part of IgG, immunoglobulin heavy chain binding protein, citrullinated proteins, and glucose 6 phosphoisomerase are detected [3]. In addition to autoantibodies, T cells activated with antigens presented on MHC play an important role in the development of C 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu 1644 Mihoko Shibuya et al. Eur. J. Immunol. 2015. 45: 1643-1653 inflammation. T cells infiltrate the synovial tissue of RA patients [4] and particular HLA-DR alleles show association with susceptibility to RA [5][6][7][8]. Developmental abnormality of T cells such as neonatal thymectomy a...

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Analysis of Autoreactive CD4 T Cells in Experimental Autoimmune Encephalomyelitis after Primary and Secondary Challenge Using MHC Class II Tetramers

Cited by 43 publications

References 47 publications

Induction of Autoimmunity by Expansion of Autoreactive CD4+CD62Llow Cells In Vivo

Induction of Autoimmunity by Expansion of Autoreactive CD4+CD62Llow Cells In Vivo

Detection of low‐frequency human antigen‐specific CD4⁺ T cells using MHC class II multimer bead sorting and immunoscope analysis

A new T‐cell activation mode for suboptimal doses of antigen under the full activation of T cells with different specificity

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Analysis of Autoreactive CD4 T Cells in Experimental Autoimmune Encephalomyelitis after Primary and Secondary Challenge Using MHC Class II Tetramers

Cited by 43 publications

References 47 publications

Induction of Autoimmunity by Expansion of Autoreactive CD4+CD62Llow Cells In Vivo

Induction of Autoimmunity by Expansion of Autoreactive CD4+CD62Llow Cells In Vivo

Detection of low‐frequency human antigen‐specific CD4+ T cells using MHC class II multimer bead sorting and immunoscope analysis

A new T‐cell activation mode for suboptimal doses of antigen under the full activation of T cells with different specificity

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Detection of low‐frequency human antigen‐specific CD4⁺ T cells using MHC class II multimer bead sorting and immunoscope analysis