Clinical-grade regulatory T cells: Comparative analysis of large-scale expansion conditions

Velaga, Sarvari; Alter, Christina; Dringenberg, Ulrike; Thiesler, Christina T.; Kuhs, Sandra; Olek, Sven; Ukena, Sya N.; Franzke, Anke

doi:10.1016/j.exphem.2016.09.008

Cited by 12 publications

(12 citation statements)

References 18 publications

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“…Regulatory T (Treg) cells present a special interest for a potential cellular therapy of allergic asthma, as they directly and specifically suppress effector Th2 cells thus lowering or even abrogating the allergen sensitization-induced tissue effects (6,(11)(12)(13). The surface and intracellular markers of Treg cells are currently well defined (11)(12)(13)(14), which permits their isolation and in vitro expansion for the subsequent use in immunotherapy (15,16). The Foxp3 molecule, which determines the Treg cell lineage commitment, has been a main focus in Treg cell detection and characterization.…”

Section: Introductionmentioning

confidence: 99%

“…The Foxp3 molecule, which determines the Treg cell lineage commitment, has been a main focus in Treg cell detection and characterization. The regulation of this molecule expression by different factors plays an important role in Treg cell differentiation, expansion, and their ability to perform suppressive activity (11)(12)(13)(14)(15)(16)(17). Studies in murine models have shown that one of the members of the neuroimmune semaphorin (Sema) family proteins, Sema4A, critically regulates the stability and function of mouse Treg cells (17,18).…”

Section: Introductionmentioning

confidence: 99%

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Semaphorin 4A Stabilizes Human Regulatory T Cell Phenotype via Plexin B1

Chapoval

Hritzo

et al. 2019

ImmunoHorizons

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We previously reported that neuroimmune semaphorin (Sema) 4A regulates the severity of experimental allergic asthma and increases regulatory T (Treg) cell numbers in vivo; however, the mechanisms of Sema4A action remain unknown. It was also reported that Sema4A controls murine Treg cell function and survival acting through neuropilin 1 (NRP-1) receptor. To clarify Sema4A action on human T cells, we employed T cell lines (HuT78 and HuT102), human PBMCs, and CD4 + T cells in phenotypic and functional assays. We found that HuT78 demonstrated a T effector-like phenotype (CD4 + CD25 low Foxp3 2), whereas HuT102 expressed a Treg-like phenotype (CD4 + CD25 hi Foxp3 +). Neither cell line expressed NRP-1. HuT102 cells expressed Sema4A counter receptor Plexin B1, whereas HuT78 cells were Sema4A +. All human peripheral blood CD4 + T cells, including Treg cells, expressed PlexinB1 and lacked both NRP-1 and-2. However, NRP-1 and Sema4A were detected on CD3 negative CD4 intermediate human monocytes. Culture of HuT cells with soluble Sema4A led to an upregulation of CD25 and Foxp3 markers on HuT102 cells. Addition of Sema4A increased the relative numbers of CD4 + CD25 + Foxp3 + cells in PBMCs and CD4 + T cells, which were NRP-1 negative but PlexinB1 + , suggesting the role of this receptor in Treg cell stability. The inclusion of anti-PlexinB1 blocking Ab in cultures before recombinant Sema4A addition significantly decreased Treg cell numbers as compared with cultures with recombinant Sema4A alone. Sema4A was as effective as TGF-b in inducible Treg cell induction from CD4 + CD25 depleted cells but did not enhance Treg cell suppressive activity in vitro. These results suggest strategies for the development of new Sema4A-based therapeutic measures to combat allergic inflammatory diseases. ImmunoHorizons, 2019, 3: 71-87.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Semaphorin 4A Stabilizes Human Regulatory T Cell Phenotype via Plexin B1

Chapoval

Hritzo

et al. 2019

ImmunoHorizons

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“…As applications of Tregs are reaching the clinic in increasing frequency, this simple modification/correction of an established method proposed here allows for the selection of pure fractions of specified Tregs or defined mixtures of such fractions for clinical studies and applications thereof (Gitelman & Bluestone, ; Mathew et al, ; Putnam et al, ; Safinia et al, ; Velaga et al, ; Wing et al, ). In this respect it is of interest that CD4CD25 high CD127 low/− CD45RA + cells were found to be better precursors for the development ex‐vivo of robust Tregs from the peripheral blood of Type 1 diabetes patients than their CD45RO + counterparts, all performed under good manufacturing practice conditions, suitable for clinical application (Gitelman & Bluestone, 2016; Putnam et al, ).…”

Section: Discussionmentioning

confidence: 99%

A modified flow cytometry method for objective estimation of human CD4⁺ regulatory T cells (CD4⁺ Tregs) in peripheral blood, via CD4/CD25/CD45RO/FoxP3 labeling

Petsiou

Paschou

Vartholomatos

et al. 2019

Cytometry Part B Clinical

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Background: Several methods exist for flow-cytometric estimation of human peripheral blood CD4 + T regulatory cells (CD4 + Tregs). Methods:We report our experience with the estimation of human CD4 + Tregs via three different characterizations using flow cytometry (CD25 high FoxP3 + , CD25 high CD127 low/− FoxP3 + , and CD4 + CD25 high/int CD45ROFoxP3 + ) in normal subjects. We have used these methods on the control populations from two studies (32 and 36 subjects, respectively), the latter two methods retrospectively on the subjects of the first study. The six CD4 + T cell fractions obtained by the third method were differentially colored to ascertain the Abbreviations: CD45RA-RO, CD45RA or CD45RO; eTregs, effector Tregs (CD4 + CD25 high CD45RA − , or CD4 + CD25 high CD45RO + ), or active Tregs (aTregs); IPEX, immune deficiencypolyendocrinopathy-enteropathy-X-linked; iTregs, induced Tregs; mAb, monoclonal antibody; MFI, mean fluorescence intensity; nTregs, naïve Tregs (CD4 + CD25 int CD45RA + , or CD4 + CD25 int CD45RO − )

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“…Accordingly, new treatments to induce tolerance in MG could be a way forward to prevent progression and possibly offer cure from disease ( 3 ). Indeed, several publications suggest that restoring a functional Treg population could be a curative therapeutic intervention in MG ( 51 – 53 ). One such approach has focused on identifying the immunodominant epitopes in the AChR to use these for protective immunizations ( 3 , 54 ).…”

Section: Discussionmentioning

confidence: 99%

A Sensitive Method for Detecting Peptide-specific CD4+ T Cell Responses in Peripheral Blood from Patients with Myasthenia Gravis

et al. 2017

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Myasthenia gravis (MG) is an autoimmune neurological disorder typified by skeletal muscle fatigue and most often production of autoantibodies against the nicotinic acetylcholine receptor (AChR). The present study was undertaken to assess the extent of AChR-peptide recognition in MG patients using co-culturing (DC:TC) of autologous monocyte-derived dendritic cells (moDCs) and highly enriched CD4+ T cells from the blood as compared to the traditional whole peripheral blood mononuclear cell (PBMC) cultures. We found that the DC:TC cultures were highly superior to the PBMC cultures for detection of reactivity toward HLA-DQ/DR-restricted AChR-peptides. In fact, whereas DC:TC cultures identified recognition in all MG patients the PBMC cultures failed to detect responsiveness in around 40% of the patients. Furthermore, reactivity to multiple peptides was evident in DC:TC cultures, while PBMC cultures mostly exhibited reactivity to a single peptide. No healthy control (HC) CD4+ T cells responded to the peptides in either culture system. Interestingly, whereas spontaneous production of IFNγ and IL-17 was observed in the DC:TC cultures from MG patients, recall responses to peptides enhanced IL-10 production in 9/13 MG patients, while little increase in IFNγ and IL-17 was seen. HCs did not produce cytokines to peptide stimulations. We conclude that the DC: TC culture system is significantly more sensitive and better identifies the extent of responsiveness in MG patients to AChR-peptides than traditional PBMC cultures.

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Clinical-grade regulatory T cells: Comparative analysis of large-scale expansion conditions

Cited by 12 publications

References 18 publications

Semaphorin 4A Stabilizes Human Regulatory T Cell Phenotype via Plexin B1

Semaphorin 4A Stabilizes Human Regulatory T Cell Phenotype via Plexin B1

A modified flow cytometry method for objective estimation of human CD4⁺ regulatory T cells (CD4⁺ Tregs) in peripheral blood, via CD4/CD25/CD45RO/FoxP3 labeling

A Sensitive Method for Detecting Peptide-specific CD4+ T Cell Responses in Peripheral Blood from Patients with Myasthenia Gravis

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Clinical-grade regulatory T cells: Comparative analysis of large-scale expansion conditions

Cited by 12 publications

References 18 publications

Semaphorin 4A Stabilizes Human Regulatory T Cell Phenotype via Plexin B1

Semaphorin 4A Stabilizes Human Regulatory T Cell Phenotype via Plexin B1

A modified flow cytometry method for objective estimation of human CD4+ regulatory T cells (CD4+ Tregs) in peripheral blood, via CD4/CD25/CD45RO/FoxP3 labeling

A Sensitive Method for Detecting Peptide-specific CD4+ T Cell Responses in Peripheral Blood from Patients with Myasthenia Gravis

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Product

Resources

About

A modified flow cytometry method for objective estimation of human CD4⁺ regulatory T cells (CD4⁺ Tregs) in peripheral blood, via CD4/CD25/CD45RO/FoxP3 labeling