Methods to Measure MDSC Immune Suppressive Activity <i>In Vitro</i> and <i>In Vivo</i>

Solito, Samantha; Pinton, Laura; Sanctis, Francesco De; Ugel, Stefano; Bronte, Vincenzo; Mandruzzato, Susanna; Marigo, Ilaria

doi:10.1002/cpim.61

Cited by 42 publications

(21 citation statements)

References 48 publications

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“…Myeloid cells were identified as described in Fig 1B and purified using FACS. Average purity of sorted samples was > 80%, which is within the normal expected range when using this method [27]. Representative dot plots of pre and post-sort samples are provided in S1 Fig.…”

Section: Physical Activity Alters the Suppressive Function Of Mdscssupporting

confidence: 63%

Physical activity delays accumulation of immunosuppressive myeloid-derived suppressor cells

et al. 2020

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Background Myeloid-derived suppressor cells (MDSCs) are potent suppressors of immune function and may play a key role in the development and progression of metastatic cancers. Aerobic exercise has been shown to have anticancer effects, yet the mechanisms behind this protection are largely unknown. Therefore, we examined the effects of physical activity on MDSC accumulation and function. Methods Female BALB/c mice were assigned to one of two primary groups: sedentary tumor (SED+ TUM) or wheel run tumor (WR+TUM). After 6 weeks of voluntary wheel running, all animals were randomly subdivided into 4 different timepoint groups; 16, 20, 24, and 28 days posttumor injection. All mice were inoculated with 4T1 mammary carcinoma cells in the mammary fat pad and WR groups continued to run for the specified time post-injection. Spleen, blood, and tumor samples were analyzed using flow cytometry to assess proportions of MDSCs. Results Cells expressing MDSC biomarkers were detected in the spleen, blood, and tumor beginning at d16. However, since there was no evidence of immunosuppressive function until d28, we refer to them as immature myeloid cells (IMCs). Compared to SED+TUM, levels of IMCs in the spleen were significantly lower (p < 0.05) in WR+TUM at day 16 (33.0 ± 5.2%; 23.1 ± 10.2% of total cells, respectively) and day 20 (33.9 ± 8.1%; 24.3 ± 5.1% of total cells, respectively). Additionally, there were fewer circulating IMCs in WR+TUM at day 16 and MDSC levels were significantly lower (p < 0.05) in the tumor at day 28 in WR+TUM. Additionally, a non-significant 62% and 26% reduction in metastatic lung nodules was observed at days 24 and 28, respectively. At day 28, MDSCs harvested from SED+TUM significantly

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Section: Physical Activity Alters the Suppressive Function Of Mdscssupporting

confidence: 63%

Physical activity delays accumulation of immunosuppressive myeloid-derived suppressor cells

et al. 2020

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“…Therefore, functional tests cannot be considered as an adjunct to MDSC identification but, on the contrary, have a prioritizing role for definition of MDSC subsets. However, since standardizing functional tests among different laboratories might represent a challenging hurdle [38], we focused our subsequent efforts in identifying genes and molecular pathways that can represent clues to their immunosuppressive ability.…”

Section: Discussionmentioning

confidence: 99%

Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3

Trovato¹,

Fiore²,

Sartori³

et al. 2019

j. immunotherapy cancer

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154

143

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Background: Pancreatic ductal adenocarcinoma (PDAC) is a highly devastating disease with an overall 5-year survival rate of less than 8%. New evidence indicates that PDAC cells release pro-inflammatory metabolites that induce a marked alteration of normal hematopoiesis, favoring the expansion and accumulation of myeloid-derived suppressor cells (MDSCs). We report here that PDAC patients show increased levels of both circulating and tumor-infiltrating MDSC-like cells. Methods: The frequency of MDSC subsets in the peripheral blood was determined by flow cytometry in three independent cohorts of PDAC patients (total analyzed patients, n = 117). Frequency of circulating MDSCs was correlated with overall survival of PDAC patients. We also analyzed the frequency of tumor-infiltrating MDSC and the immune landscape in fresh biopsies. Purified myeloid cell subsets were tested in vitro for their T-cell suppressive capacity. Results: Correlation with clinical data revealed that MDSC frequency was significantly associated with a shorter patients' overall survival and metastatic disease. However, the immunosuppressive activity of purified MDSCs was detectable only in some patients and mainly limited to the monocytic subset. A transcriptome analysis of the immunosuppressive M-MDSCs highlighted a distinct gene signature in which STAT3 was crucial for monocyte reprogramming. Suppressive M-MDSCs can be characterized as circulating STAT3/arginase1-expressing CD14 + cells. Conclusion: MDSC analysis aids in defining the immune landscape of PDAC patients for a more appropriate diagnosis, stratification and treatment.

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“…T cell co-culture was performed as previously described 67 with minor modifications. Briefly, tissue-culture plates were coated with 5 μg/mL purified anti-CD3 (clone HIT3a, Biolegend) at 4°C overnight and subsequently washed twice with 1X PBS.…”

Section: T Cell Co-culture and Proliferation Assaymentioning

confidence: 99%

Induction of a regulatory myeloid program in bacterial sepsis and severe COVID-19

Reyes

Filbin

Bhattacharyya

et al. 2020

Preprint

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A recent estimate suggests that one in five deaths globally are associated with sepsis1. To date, no targeted treatment is available for this syndrome, likely due to substantial patient heterogeneity2,3 and our lack of insight into sepsis immunopathology4. These issues are highlighted by the current COVID-19 pandemic, wherein many clinical manifestations of severe SARS-CoV-2 infection parallel bacterial sepsis5–8. We previously reported an expanded CD14+ monocyte state, MS1, in patients with bacterial sepsis or non-infectious critical illness, and validated its expansion in sepsis across thousands of patients using public transcriptomic data9. Despite its marked expansion in the circulation of bacterial sepsis patients, its relevance to viral sepsis and association with disease outcomes have not been examined. In addition, the ontogeny and function of this monocyte state remain poorly characterized. Using public transcriptomic data, we show that the expression of the MS1 program is associated with sepsis mortality and is up-regulated in monocytes from patients with severe COVID-19. We found that blood plasma from bacterial sepsis or COVID-19 patients with severe disease induces emergency myelopoiesis and expression of the MS1 program, which are dependent on the cytokines IL-6 and IL-10. Finally, we demonstrate that MS1 cells are broadly immunosuppressive, similar to monocytic myeloid-derived suppressor cells (MDSCs), and have decreased responsiveness to stimulation. Our findings highlight the utility of regulatory myeloid cells in sepsis prognosis, and the role of systemic cytokines in inducing emergency myelopoiesis during severe bacterial and SARS-CoV-2 infections.

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Methods to Measure MDSC Immune Suppressive Activity In Vitro and In Vivo

Cited by 42 publications

References 48 publications

Physical activity delays accumulation of immunosuppressive myeloid-derived suppressor cells

Physical activity delays accumulation of immunosuppressive myeloid-derived suppressor cells

Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3

Induction of a regulatory myeloid program in bacterial sepsis and severe COVID-19

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