Phenotypic Characterization and Isolation of Myeloid‐Derived Suppressor Cells

Reuven, Or; Mikula, I.; Ashkenazi‐Preiser, Hadas; Twaik, Nira; Ben‐Meir, Kerem; Meirow, Yaron; Daniel, Leonor; Kariv, Guy; Kurd, Mahdi; Baniyash, Michal

doi:10.1002/cpz1.561

Cited by 3 publications

(23 citation statements)

References 12 publications

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“…Control and chronically inflamed mice Splenocyte single-cell suspension (see companion Current Protocols article by Reuven et al, 2022) 25-G needles 1-ml syringes Mouse restrainer for tail vein injection Additional reagents and equipment for mouse euthanasia (see Current Protocols article by Donovan & Brown, 2005) CTL adoptive transfer 1. Obtain CTLs as described in Basic Protocol 4.…”

Section: Additional Materials (Also See Basic Protocol 4)mentioning

confidence: 99%

“…Select methods describe manipulation of the entire splenocyte population or of isolated T cells as described here (also see a companion Current Protocols article by Reuven et al, 2022). When harvesting other tissue, use appropriate tissue-specific methods, as additional use of enzymatic digestion and/or mechanical force might be required.…”

Section: Evaluating Mdsc Suppressive Functionmentioning

confidence: 99%

“…1. Acquire splenocyte single-cell suspension (see companion Current Protocols article by Reuven et al, 2022). Resuspend cells in RPMI 1640 full medium.…”

Section: Sample Preparation and Platingmentioning

confidence: 99%

“…This protocol can be adapted for both isolated T cells and single‐cell suspensions of splenocytes acquired using Basic Protocol 2 in this manuscript and in protocols presented in a companion Current Protocols article by Reuven et al. (2022).…”

Section: Introductionmentioning

confidence: 99%

“…Herein, we describe methods for the analysis of MDSC immunosuppressive features and functions, measuring different mediators that contribute to their activities and how they impact on T cell function. The protocols described are a continuation to those in a companion Current Protocols article by Reuven et al (2022), which uses a generated single-cell suspension and isolated cells to test their activity.…”

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confidence: 99%

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Functional Assays Evaluating Immunosuppression Mediated by Myeloid‐Derived Suppressor Cells

Reuven¹,

Mikula²,

Ashkenazi‐Preiser³

et al. 2022

Current Protocols

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Myeloid-derived suppressor cells (MDSCs) are heterogenous populations of immature myeloid cells that can be divided into two main subpopulations, polymorphonuclear (PMN) MDSCs and monocytic (M) MDSCs. These cells accumulate during chronic inflammation, characterizing an array of pathologies such as cancer, inflammatory bowel disease, and infectious and autoimmune diseases, and induce immunosuppression. The suppressive effects of MDSCs on the immune system are studied mainly when focusing on their features, functions, and impact on target cells such as T cells, natural killer cells, and B cells, among others. Herein, we describe methods for the analysis of MDSC immunosuppressive features and functions, measuring different mediators that contribute to their activities and how they impact on T cell function. The protocols described are a continuation to those in a companion Current Protocols article by Reuven et al. (2022), which uses a generated single-cell suspension and isolated cells to test their activity.

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Section: Additional Materials (Also See Basic Protocol 4)mentioning

confidence: 99%

Section: Evaluating Mdsc Suppressive Functionmentioning

confidence: 99%

“…1. Acquire splenocyte single-cell suspension (see companion Current Protocols article by Reuven et al, 2022). Resuspend cells in RPMI 1640 full medium.…”

Section: Sample Preparation and Platingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Functional Assays Evaluating Immunosuppression Mediated by Myeloid‐Derived Suppressor Cells

Reuven¹,

Mikula²,

Ashkenazi‐Preiser³

et al. 2022

Current Protocols

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An In Vivo Mouse Model for Chronic Inflammation–Induced Immune Suppression: A “Factory” for Myeloid‐Derived Suppressor Cells (MDSCs)

Ben‐Meir¹,

Twaik²,

Meirow³

et al. 2022

Current Protocols

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Myeloid-derived suppressor cells (MDSCs) represent a heterogeneous population of immature myeloid cells known to play a role in perpetuating a wide range of pathologies, such as chronic infections, autoimmune diseases, and cancer. MDSCs were first identified in mice by the markers CD11b + Gr1 + , and later, based on their morphology, they were classified into two subsets: polymorphonuclear MDSCs, identified by the markers CD11b + Ly6G + Ly6C Low , and monocytic MDSCs, detected as being CD11b + Ly6G -Ly6C Hi . MDSCs are studied as immunosuppressive cells in various diseases characterized by chronic inflammation and are associated with disease causes/triggers such as pathogens, autoantigens, and cancer. Therefore, different diseases may diversely affect MDSC metabolism, migration, and differentiation, thus influencing the generated MDSC functional features and ensuing suppressive environment. In order to study MDSCs in a pathology-free environment, we established and calibrated a highly reproducible mouse model that results in the development of chronic inflammation, which is the major cause of MDSC accumulation and immune suppression. The model presented can be used to study MDSC phenotypes, functional diversity, and plasticity. It also permits study of MDSC migration from the bone marrow to peripheral lymphatic and non-lymphatic organs and MDSC crosstalk with extrinsic factors, both in vivo and ex vivo. Furthermore, this model can serve as a platform to assess the effects of anti-MDSC modalities.

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Role of myeloid-derived suppressor cells in prediction of the effectiveness of biologics in children with psoriasis

Kuptsova,

Radygina,

Freidlin

et al. 2023

Russian Journal of Immunology

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Psoriasis is a chronic inflammatory skin disease characterized by increased proliferation of epidermal cells, impaired keratinization, and an inflammatory reaction in the dermis due to activation of T-lymphocytes and synthesis of proinflammatory cytokines. Pathophysiology of psoriasis is associated not only with activation of proinflammatory reactions, but also with decreased anti-inflammatory functions of immunosuppressive cells. It is known that Treg, Breg and MDSCs do not perform their classical homeostatic functions in psoriasis. In recent years, there have been more and more cases of developing resistance to ongoing therapy with genetically engineered biological drugs (GEBD) in childhood, requiring replacement or discontinuation of the drug. The aim of our work was to estimate the content of MDSCs subpopulations and their functional activity in the peripheral blood of children with psoriasis at different effectiveness of biotherapeutic drugs. We examined 110 children with psoriasis vulgaris before the appointment of biologics, at 16 and 52 weeks of therapy with adalimumab, etanercept and ustikinumab, aged 6 to 18 years. Сomparison group consisted of 34 healthy children. The effectiveness of therapy was assessed by the achievement of PASI 75 by one year of therapy. Contents of myeloid-derived suppressor cells (MDSCs) and their subpopulations, and the activity of arginase-1 were assessed by multicolor flow cytometry. An increased content of MDSCs was found in children with psoriasis against the comparison group (p = 0.000). Analysis of the effectiveness of biologics in children with psoriasis, according to PASI, showed a significant reduction in disease severity in the group of patients with good effect, both at week 16 of therapy (p = 0.000) and by one year (p = 0.017). In the group of patients with good effect of biological therapy, percentage of total MDSCs population was higher, both before start of treatment and by 52nd week of therapy (p 0.01). Children with psoriasis showed increased immunosuppressive function of MDSCs by arginase-1 activity versus the comparison group (p = 0.000). The arginase-1 activity in patients with psoriasis at the stage of disease regression (PASI 10) was significantly increased relative to children in progressive stage of psoriasis (PASI10; p = 0.001). Thus, the content of MDSCs and their suppressive activity in children with psoriasis is an informative efficiency predictor of the biological drugs. Fading of biotherapy effect after the induction course is accompanied by decreased number of MDSCs and their functional activity.

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Phenotypic Characterization and Isolation of Myeloid‐Derived Suppressor Cells

Cited by 3 publications

References 12 publications

Functional Assays Evaluating Immunosuppression Mediated by Myeloid‐Derived Suppressor Cells

Functional Assays Evaluating Immunosuppression Mediated by Myeloid‐Derived Suppressor Cells

An In Vivo Mouse Model for Chronic Inflammation–Induced Immune Suppression: A “Factory” for Myeloid‐Derived Suppressor Cells (MDSCs)

Role of myeloid-derived suppressor cells in prediction of the effectiveness of biologics in children with psoriasis

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