A rapid diagnostic test for human regulatory T-cell function to enable regulatory T-cell therapy

Canavan, James B.; Afzali, Behdad; Scottà, Cristiano; Fazekasova, Henrieta; Edozie, Francis C.; MacDonald, Thomas T.; Hernandez-Fuentes, Maria P.; Lombardi, Giovanna; Lord, Graham M.

doi:10.1182/blood-2011-09-380048

Cited by 72 publications

(79 citation statements)

References 39 publications

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“…The FastImmune Regulatory T-Cell Function Kit (BD) was used in combination with CD8 (SK1) and Live/Dead Aqua for the suppression assay, which was performed as described with some modification; the suppression of activation markers expression can also predict the proliferative suppression (31). Autologous PBMCs were used as responder cells; they were cultured either alone or with the Treg subset at a ratio of 1 Treg to 4 responders, in complete medium.…”

Section: Suppression Assay Of Early Activation and Cytokines Productionmentioning

confidence: 99%

Human CD45RA− FoxP3hi Memory-Type Regulatory T Cells Show Distinct TCR Repertoires With Conventional T Cells and Play an Important Role in Controlling Early Immune Activation

Lei

Kuchenbecker

Streitz

et al. 2015

American Journal of Transplantation

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Section: Suppression Assay Of Early Activation and Cytokines Productionmentioning

confidence: 99%

Human CD45RA− FoxP3hi Memory-Type Regulatory T Cells Show Distinct TCR Repertoires With Conventional T Cells and Play an Important Role in Controlling Early Immune Activation

Lei

Kuchenbecker

Streitz

et al. 2015

American Journal of Transplantation

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“…We show that Zap70 phosphorylation occurs within few minutes of Tresp stimulation and is detected for only 10 minutes, similar to the short period of Zap70 phosphorylation previously reported [11]. Treg were reported to require several hours for exerting their effect on Tresp [9], therefore we employed a "2-stages approach" for activating Treg and then measuring Zap70 phosphorylation in Tresp using anti-CD3 and anti-CD28 antibodies for both stages. Whether alternative methods, such as initial activation of Tresp with PMA and ionomycin that act downstream of Zap70, followed by stimulation of the T cell synapse through CD3 and CD28, IL2 or other T cell stimulators, will produce similar or more robust responses than observed here, will need to be studied.…”

Section: Discussionmentioning

confidence: 94%

“…Similarly, IL2 or interferon-gamma production by Tresp, CFSE dilution and [ 3 H]-Thymidine incorporation have been employed to measure the response of activated Tresp [8]. Suppression of Tresp up-regulation of CD154 (the ligand for CD40) and CD69 that can be detected 6-24 hours after activation has been shown to correlate well with the gold standard suppressor cell assays [9]. Zeta-associated protein (Zap)70 is a tyrosine kinase that is central for human and murine T-cell development and activation [10].…”

Section: Introductionmentioning

confidence: 99%

“…Tresp (CD4 + CD25 -) were isolated similarly from the spleens of ADAproficient mice only, as ADA-deficient Tresp have reduced stimulation responses [12]. Tresp were stimulated with 5 µg/ml of anti-CD3 and anti-CD28 antibodies, as described previously [9]. Zap70 phosphorylation (following fixation and permeabilization) and surface CD69 expression were determined by flow cytometry (using the FACSCalibur, Becton-Dickinson, San Jose, CA, USA) at the indicated times.…”

Section: Cells Isolation Stimulation Activation Suppression and Anmentioning

confidence: 99%

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Regulatory T cells Suppress Zap70 Phosphorylation in Responding T cells

Faitelson

Min²,

Grunebaum³

2016

J Clin Cell Immunol

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Objective: Regulatory T cells (Treg) have a critical role in controlling responding T cells (Tresp), thereby preventing autoimmune manifestations including those observed in adenosine deaminase (ADA) enzyme-deficient patients and mice (ADA-KO). Treg suppress various functions of Tresp such as proliferation, cytokine production and expression of activation markers, yet it is not known if Treg suppress the early steps of Tresp activation, such as the phosphorylation of the Zeta-associated protein (Zap)70.Methods: Zap70 phosphorylation and CD69 expression in anti-CD3 and anti-CD28 stimulated mice CD4 + CD25 -Tresp were measured by flow cytometry in the presence or absence of CD4 + CD25 + Treg activated with anti-CD3 and anti-CD28 antibodies. Suppression of Zap70 phosphorylation in Tresp from normal mice cultured with Treg from ADA-KO mice, either treated with PEG-ADA enzyme replacement or untreated, was similarly measured.Results: Zap70 phosphorylation in activated Tresp was markedly (50 ± 13%) decreased 2 hours after culture with Treg, while 51 ± 8% suppression of CD69 expression was only detected after 7 hours. Suppression of Zap70 phosphorylation in activated Tresp correlated with the ratio of Treg to Tresp. Treg from ADA-KO mice had significantly reduced (p=0.012) ability to suppress Zap70 phosphorylation (16.2 ± 16.7%) compared to Treg from healthy littermates (51.6 ± 23.4%), while PEG-ADA treatment restored Treg suppressive ability (45 ± 10%). Conclusions:Treg suppress Zap70 phosphorylation in Tresp, a finding that might help better understand and assess Treg function.

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“…All other markers, including CTLA-4, CD39, CCR7, HELIOS or CD69 were considered as "optional." Inhibition of responder T-cell proliferation or T-cell activation was considered to be a useful functional assay [19,20]. Ki67, the universal proliferation marker, proved to be useful as functional Treg marker but only in vivo, as Treg in mice were shown to proliferate vigorously and express Ki67 [21].…”

Section: Measurements Of Human Tregmentioning

confidence: 99%

Clinical Impact of Regulatory T cells (Treg) in Cancer and HIV

Whiteside

2014

Cancer Microenvironment

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The role of regulatory T cells, (Treg) in human cancer and HIV-1 infections has been under intense scrutiny. While the lack of a marker specific for human Treg has made it challenging to phenotype these cells, combinations of several markers and functional attributes of Treg have made it possible to assess their contributions to immune homeostasis in health and disease. Treg diversity and their plasticity create a challenge in deciding whether they are beneficial to the host by down-regulating excessive immune activation or are responsible for adverse effects such as suppression of anti-tumor immune responses resulting in promotion of tumor growth. Treg are emerging as active participants in several biochemical pathways involved in immune regulation. This review attempts to integrate current information about human Treg in respect to their activities in cancer and HIV-1. The goal is to evaluate the potential of Treg as targets for future immune or pharmacologic therapies for cancer or HIV-1 infections.

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A rapid diagnostic test for human regulatory T-cell function to enable regulatory T-cell therapy

Abstract: Regulatory T cells (CD4 ؉CD25

Cited by 72 publications

References 39 publications

Human CD45RA− FoxP3hi Memory-Type Regulatory T Cells Show Distinct TCR Repertoires With Conventional T Cells and Play an Important Role in Controlling Early Immune Activation

Human CD45RA− FoxP3hi Memory-Type Regulatory T Cells Show Distinct TCR Repertoires With Conventional T Cells and Play an Important Role in Controlling Early Immune Activation

Regulatory T cells Suppress Zap70 Phosphorylation in Responding T cells

Clinical Impact of Regulatory T cells (Treg) in Cancer and HIV

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