Isolation of human antigen‐specific regulatory T cells with high suppressive function

Noyan, Fatih; Lee, Young-Seon; Zimmermann, Katharina; Hardtke‐Wolenski, Matthias; Taubert, Richard; Warnecke, G.; Knoefel, A.; Schulde, E.; Olek, Sven; Manns, Michael P.; Jaeckel, Elmar

doi:10.1002/eji.201344381

Cited by 45 publications

(43 citation statements)

References 41 publications

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“…The expression of IL-10 transcript, together with the relative increase in Lrrc32 and Foxp3 transcripts in the CD154 − population compared to the CD154 + population (Figure 3D and 4C), was consistent with previous reports showing that regulatory T cells lack pre-formed CD154 and that induced regulatory T cells are unlikely to be included in the CD154 + population [43–45]. …”

Section: Resultssupporting

confidence: 91%

“…Expression microarray analysis performed on mycobacteria-specific CD4 + T cells further supported the validity of using CD154 expression as a surrogate marker for antigen-specificity. In addition, microarray analysis revealed that while multiple functional T helper subsets were present in the CD154 + population, antigen-specific regulatory T cells were likely to be excluded, consistent with previous reports in the literature [43–45]. …”

Section: Discussionsupporting

confidence: 88%

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Transcriptome Analysis of Mycobacteria-Specific CD4+ T Cells Identified by Activation-Induced Expression of CD154

Kunnath-Velayudhan

Goldberg

Saini

et al. 2017

The Journal of Immunology

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Analysis of antigen-specific CD4+ T cells in mycobacterial infections at the transcriptome level is informative but technically challenging. While several methods exist for identifying antigen-specific T cells, including intracellular cytokine staining, cell surface cytokine-capture assays, and staining with peptide:MHC-II multimers, all of these have significant technical constraints that limit their usefulness. Measurement of activation-induced expression of CD154 has been reported to detect live, antigen-specific CD4+ T cells but this approach remains underexplored, and to our knowledge has not previously been applied in mycobacteria-infected animals. Here we show that CD154 expression identifies adoptively transferred or endogenous antigen-specific CD4+ T cells induced by Mycobacterium bovis Bacillus Calmette-Guérin (BCG) vaccination. We confirmed that antigen-specific cytokine production was positively correlated with CD154 expression by CD4+ T cells from BCG-vaccinated mice, and show that high quality microarrays can be performed from RNA isolated from CD154+ cells purified by cell sorting. Analysis of microarray data demonstrated that the transcriptome of the CD4+ CD154+ cells was distinct from that of CD154− cells, and showed major enrichment of transcripts encoding multiple cytokines and pathways of cellular activation. One notable finding was the identification of a previously unrecognized subset of mycobacteria-specific CD4+ T cells characterized by the production of interleukin-3. Our results support the use of CD154 expression as a practical and reliable method to isolate live, antigen-specific CD4+ T cells for transcriptomic analysis and potentially for a range of other studies in infected or previously immunized hosts.

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Section: Resultssupporting

confidence: 91%

Section: Discussionsupporting

confidence: 88%

Transcriptome Analysis of Mycobacteria-Specific CD4+ T Cells Identified by Activation-Induced Expression of CD154

Kunnath-Velayudhan

Goldberg

Saini

et al. 2017

The Journal of Immunology

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“…Furthermore, it has been demonstrated that Hsp90-mediated Hsp70 induction is necessary for promoting T reg survival and suppressive function. Strikingly, CD4 + CD25 + LAP + iT regs demonstrated high suppressive and antigen-specific capacity, substantiating a recent report describing LAP + expression on CD4 T cells as a reliable marker for the identification of antigen-specific T regs [28]. Furthermore, Mahalingam et al [67] demonstrated that isolated CD4 + CD25 + LAP + showed higher suppressive activity than CD4 + CD25 + LAP 2 , underscoring the use of LAP expression for identification of a potent, suppressive subgroup of T regs .…”

Section: Discussionsupporting

confidence: 80%

Combined PI3K/Akt and Hsp90 targeting synergistically suppresses essential functions of alloreactive T cells and increases Tregs

Berges

Bedke

Stuehler

et al. 2015

Journal of Leukocyte Biology

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Acute graft-versus-host disease is still a major cause of transplant-related mortality after allogeneic stem cell transplantation. It requires immunosuppressive treatments that broadly abrogate T cell responses, including beneficial ones directed against tumor cells or infective pathogens. Inhibition of the heat shock protein of 90 kDa has been demonstrated to eliminate tumor cells, as well as alloreactive T cells while preserving antiviral T cell immunity. Here, we show that the suppressive effects of heat shock protein of 90 kDa inhibition on alloreactive T cells were synergistically enhanced by concomitant inhibition of the PI3K/Akt signaling pathway, which is also strongly activated upon allogeneic stimulation. Molecular analyses revealed that this antiproliferative effect was mainly mediated by induction of cell-cycle arrest and apoptosis. In addition, we observed an increased proportion of activated regulatory T cells, which critically contribute to acute graft-versus-host disease control, upon combined heat shock protein of 90 kDa/Akt isoforms 1 and 2 or heat shock protein of 90 kDa/PI3K/p110δ isoform inhibition. Moreover, antiviral T cell immunity was functionally preserved after combined heat shock protein of 90 kDa/Akt isoforms 1 and 2 inhibition. Taken together, our data suggest that the combined heat shock protein of 90 kDa/PI3K/Akt inhibition approach represents a reasonable dual strategy to suppress residual tumor growth and efficiently deplete alloreactive T cells and thus, provide a rationale to prevent and treat acute graft-versus-host disease selectively without impairing pathogen-specific T cell immunity.

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“…In recent years, GARP is discovered to express highly on the surface of activated human Tregs in vitro [36]. Freshly isolated human Tregs and Tregs clones express a low level of surface GARP.…”

Section: Treg Markersmentioning

confidence: 99%

GARP: a surface molecule of regulatory T cells that is involved in the regulatory function and TGF-β releasing

Sun

Hao

2016

Oncotarget

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There are many molecules that define regulatory T cells (Tregs) phenotypically and functionally. Glycoprotein A repetitions predominant (GARP) is a transmembrane protein containing leucine rich repeats. Recently, GARP is found to express highly on the surface of activated Tregs. The combination of GARP and other surface molecules isolates Tregs with higher purity. Besides, GARP is a cell surface molecule of Tregs that maintains their regulatory function and homeosatsis. GARP has also been proved to promote the activation and secretion of transforming growth factor β (TGF-β). Moreover, its potential value in cancer immunotherapy is also discussed in this work.

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Isolation of human antigen‐specific regulatory T cells with high suppressive function

Cited by 45 publications

References 41 publications

Transcriptome Analysis of Mycobacteria-Specific CD4+ T Cells Identified by Activation-Induced Expression of CD154

Transcriptome Analysis of Mycobacteria-Specific CD4+ T Cells Identified by Activation-Induced Expression of CD154

Combined PI3K/Akt and Hsp90 targeting synergistically suppresses essential functions of alloreactive T cells and increases Tregs

GARP: a surface molecule of regulatory T cells that is involved in the regulatory function and TGF-β releasing

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