Identification and isolation of murine antigen‐reactive T cells according to CD154 expression

Kirchhoff, Dennis; Frentsch, Marco; Leclerk, Patrick; Bumann, Dirk; Rausch, Sebastian; Hartmann, Susanne; Thiel, Andreas; Scheffold, Alexander

doi:10.1002/eji.200737322

Cited by 55 publications

(70 citation statements)

References 29 publications

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“…Anti-CD40 mAb (BD Biosciences) and dialyzed anti-CD154-PE (BD Biosciences) were added to each culture (20 mg/ml each mAb) and incubated overnight at 37˚C. Anti-CD154 was included in the cultures to stabilize CD154 expression on the surface of recently activated CD4 + cells (21)(22)(23). Cells were stained with anti-CD69-FITC, anti-CD154-PE, anti-B220-allophycocyanin, anti-CD11b-allophycocyanin, 7-aminoactinomycin D, anti-CD25-PE-Cy7, and anti-CD4-allophycocyanin-Cy7 (all from BD Biosciences) and analyzed by flow cytometry using a FACSCanto (BD Biosciences).…”

Section: Ex Vivo Ag Recall Assaymentioning

confidence: 99%

EC144, a Synthetic Inhibitor of Heat Shock Protein 90, Blocks Innate and Adaptive Immune Responses in Models of Inflammation and Autoimmunity

Yun¹,

Harning²,

Giza³

et al. 2011

The Journal of Immunology

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Heat shock protein 90 (Hsp90) is a molecular chaperone involved in folding and stabilizing multiple intracellular proteins that have roles in cell activation and proliferation. Many Hsp90 client proteins in tumor cells are mutated or overexpressed oncogenic proteins driving cancer cell growth, leading to the acceptance of Hsp90 as a potential therapeutic target for cancer. Because several signal transduction molecules that are dependent on Hsp90 function are also involved in activation of innate and adaptive cells of the immune system, we investigated the mechanism by which inhibiting Hsp90 leads to therapeutic efficacy in rodent models of inflammation and autoimmunity. EC144, a synthetic Hsp90 inhibitor, blocked LPS-induced TLR4 signaling in RAW 264.7 cells by inhibiting activation of ERK1/2, MEK1/2, JNK, and p38 MAPK but not NF-κB. Ex vivo LPS-stimulated CD11b+ peritoneal exudate cells from EC144-treated mice were blocked from phosphorylating tumor progression locus 2, MEK1/2, and ERK1/2. Consequently, EC144-treated mice were resistant to LPS administration and had suppressed systemic TNF-α release. Inhibiting Hsp90 also blocked in vitro CD4+ T cell proliferation in mouse and human MLRs. In vivo, semitherapeutic administration of EC144 blocked disease development in rat collagen-induced arthritis by suppressing the inflammatory response. In a mouse collagen-induced arthritis model, EC144 also suppressed disease development, which correlated with a suppressed Ag-specific Ab response and a block in activation of Ag-specific CD4+ T cells. Our results describe mechanisms by which blocking Hsp90 function may be applicable to treatment of autoimmune diseases involving inflammation and activation of the adaptive immune response.

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Section: Ex Vivo Ag Recall Assaymentioning

confidence: 99%

EC144, a Synthetic Inhibitor of Heat Shock Protein 90, Blocks Innate and Adaptive Immune Responses in Models of Inflammation and Autoimmunity

Yun¹,

Harning²,

Giza³

et al. 2011

The Journal of Immunology

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“…2D) [19]. LTbR-deficient mice generated antigenspecific T cells faster than their splenectomized counterparts, presumably because they exploited the spleen for T-cell priming.…”

Section: Cells Likely Reflect the Higher Bacterial Loads Inmentioning

confidence: 99%

Secondary lymphoid organs are dispensable for the development of T‐cell‐mediated immunity during tuberculosis

et al. 2010

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Tuberculosis causes 2 million deaths per year, yet in most cases the immune response successfully contains the infection and prevents disease outbreak. Induced lymphoid structures associated with pulmonary granuloma are observed during tuberculosis in both humans and mice and could orchestrate host defense. To investigate whether granuloma perform lymphoid functions, mice lacking secondary lymphoid organs (SLO) were infected with Mycobacterium tuberculosis (MTB). As in WT mice, granuloma developed, exponential growth of MTB was controlled, and antigen-specific T-cell responses including memory T cells were generated in the absence of SLO. Moreover, adoptively transferred T cells were primed locally in lungs in a granuloma-dependent manner. T-cell activation was delayed in the absence of SLO, but resulted in a normal development program including protective subsets and functional recall responses that protected mice against secondary MTB infection. Our data demonstrate that protective immune responses can be generated independently of SLO during MTB infection and implicate local pulmonary T-cell priming as a mechanism contributing to host defense.Key words: Granuloma . Lymphoid neogenesis . T-cell development . Tuberculosis Supporting Information available online IntroductionMycobacterium tuberculosis (MTB), the causative agent of tuberculosis (TB), currently persists in one-third of the world's population [1]. In most cases, the immune response generated upon exposure contains, but does not eradicate, the bacilli and an asymptomatic persistent infection develops, termed latent TB. Approximately one in ten latently infected individuals undergo reactivation to active TB disease during their lifetime. However, Ã These authors contributed equally to this work. ÃÃ These authors share equal senior authorship. [2,3]. Granuloma are considered central for control of MTB as loss of granuloma structure is associated with poor disease outcome in humans and animal models [3,4]. Yet, the precise mechanisms by which granuloma mediate control of MTB are poorly understood. Induction of an adaptive immune response is essential for host protection against MTB. Antigen-specific T-cell responses are generated within secondary lymphoid organs (SLO), such as lymph nodes (NO) and spleen. SLO maximize efficiency of encounters between APC and naïve T lymphocytes [5]. Therefore, SLO are critical for the generation of adaptive immunity. Ectopic accumulations of lymphoid cells, known as tertiary lymphoid organs, sometimes arise during chronic inflammation or infection. Tertiary lymphoid organs resemble SLO in structure and cell composition and evidence exists that some are capable of generating adaptive immunity [5,6]. However, whether immune responses generated outside of SLO are truly effective and thus beneficial in defense against pathogens is unclear.Recent reports have described lymphoid features associated with murine and human granuloma, such as B-cell follicles and the peripheral node addressin, which is expressed on high endothel...

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“…ϩ T cells within the T eff and T reg cell populations, we examined the expression of CD154 as a marker of antigenspecific CD4 ϩ T-cell activation (15,26). As the detection of CD154 directly ex vivo is not possible, we used an optimized in vitro protocol (26) (Fig.…”

Section: Foxp3mentioning

confidence: 99%

“…As the detection of CD154 directly ex vivo is not possible, we used an optimized in vitro protocol (26) (Fig. 5A and B).…”

Section: Foxp3mentioning

confidence: 99%

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Functional Analysis of Effector and Regulatory T Cells in a Parasitic Nematode Infection

et al. 2008

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Identification and isolation of murine antigen‐reactive T cells according to CD154 expression

Cited by 55 publications

References 29 publications

EC144, a Synthetic Inhibitor of Heat Shock Protein 90, Blocks Innate and Adaptive Immune Responses in Models of Inflammation and Autoimmunity

EC144, a Synthetic Inhibitor of Heat Shock Protein 90, Blocks Innate and Adaptive Immune Responses in Models of Inflammation and Autoimmunity

Secondary lymphoid organs are dispensable for the development of T‐cell‐mediated immunity during tuberculosis

Functional Analysis of Effector and Regulatory T Cells in a Parasitic Nematode Infection

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