Detection Of Myeloid-derived Suppressor Cells Research Articles

Early Detection of Myeloid-Derived Suppressor Cells in the Lung Pre-Metastatic Niche by Shortwave Infrared Nanoprobes.

Metastatic breast cancer remains a significant source of mortality amongst breast cancer patients and is generally considered incurable in part due to the difficulty in detection of early micro-metastases. The pre-metastatic niche (PMN) is a tissue microenvironment that has undergone changes to support the colonization and growth of circulating tumor cells, a key component of which is the myeloid-derived suppressor cell (MDSC). Therefore, the MDSC has been identified as a potential biomarker for PMN formation, the detection of which would enable clinicians to proactively treat metastases. However, there is currently no technology capable of the in situ detection of MDSCs available in the clinic. Here, we propose the use of shortwave infrared-emitting nanoprobes for the tracking of MDSCs and identification of the PMN. Our rare-earth albumin nanocomposites (ReANCs) are engineered to bind the Gr-1 surface marker of murine MDSCs. When delivered intravenously in murine models of breast cancer with high rates of metastasis, the targeted ReANCs demonstrated an increase in localization to the lungs in comparison to control ReANCs. However, no difference was seen in the model with slower rates of metastasis. This highlights the potential utility of MDSC-targeted nanoprobes to assess PMN development and prognosticate disease progression.

Detection of myeloid-derived suppressor cells by flow cytometry.

Recently discovered heterogeneous myeloid-derived suppressor cells (MDSCs) are some of the most discussed immunosuppressive cells in contemporary immunology, especially in the tumor microenvironment, and are defined primarily by their T cell immunosuppressive function. The importance of these cells extend to other chronic pathological conditions as well, including chronic infection, inflammation, and tissue remodeling. In many of these conditions, their accumulation/expansion correlates with disease progression, poor prognosis, and reduced survival, which highlights the potential of how these cells may be used in a clinical setting as both prognostic factor and therapeutic target. In healthy individuals, these cells are usually not present in the circulation. Therefore, monitoring this cell population is of potential clinical significance, and utility in basic research. However, these cells have a complex phenotype without one single marker of sufficient specificity for their identification. Flow cytometry is a powerful tool allowing multi-parameter analysis of heterogeneous cell populations, which makes it ideally suitable for the complex phenotypic analysis essential for identification and enumeration of circulating MDSCs. This approach has the potential to provide a novel clinically useful tool for assessment of prognosis and treatment outcomes. The protocol in this chapter describes a flow cytometric analysis to identify and quantify MDSCs from human or mouse whole blood leukocytes and peripheral blood mononuclear cells, as well as a single cell suspension from solid tissue, by using multicolor fluorescence-conjugated antibodies against their surface markers.

Preliminary study on the significance of pathological characteristics analysis of breast cancer and detection of peripheral blood myeloid-derived suppressor cells in the evaluation of biological characteristics

ABSTRACT To explore the significance of the analysis of pathological characteristics of breast cancer and the detection of myeloid-derived suppressor cell (MDSC) levels in peripheral blood for the evaluation of biological characteristics. 138 breast cancer patients were enrolled as the research group, while 138 patients with benign breast diseases were included as the control group. All patients underwent pathological analysis and detection of peripheral blood MDSCs levels, progesterone receptor (PR), estrogen receptor (ER), human epidermal growth factor receptor 2 (HER-2), and proliferating cell nuclear antigen (Ki-67). A factorial study of stage I, II, and III breast cancer patients showed significant differences in clinicopathological characteristics, including age, tumor size, lymph node metastases, histological grading, Neuropsychiatric Inventory (NPI) score, pathological type, and family history (P < 0.05). The research group had higher levels of peripheral blood MDSCs and different cell surface markers compared to the control group (P<0.05). Positive expression of biological molecules in breast cancer, such as PR, ER, HER-2, and Ki-67, had significant differences based on lymph node metastasis and tumor size (P < 0.05). The quality of survival scores was higher in stages I and II compared to stage III (P < 0.05). Age, recurrence, metastasis, and other pathological characteristics of breast cancer have a direct impact on clinical outcomes and survival rates. Peripheral blood levels of MDSCs and other cell surface markers are significantly elevated, serving as a crucial benchmark for the subsequent evaluation of breast cancer progression.

Myeloid-derived suppressor cells level and MUC1 expression in de novo acute myeloid leukemia

AbstractBackgroundAcute myeloid leukemia (AML) is the most common acute leukemia occurring in adults. It is an aggressive myeloid neoplasm with maturation arrest of myelopoiesis, leading to an accumulation of myeloblasts in the bone marrow and peripheral blood.ObjectiveTo evaluate alterations in myeloid-derived suppressor cells level and MUC1 gene expression in patients with de novo acute myeloid leukemia concerning disease characteristics and response to induction chemotherapy.Patients and methodsThe study was performed on 50 AML patients and 50 healthy controls. Detection of myeloid-derived suppressor cells (MDSCs) in peripheral blood was performed by mononuclear separation and flow cytometry. MUC1 gene expression was performed by RNA extraction, reverse transcription, and real-time PCR at Hematology Department Medical Research Institute, Alexandria University.ResultsWe have demonstrated that AML patients had both increased presence of MDSCs in peripheral blood as well as MUC1 overexpression in comparison to normal controls. MDSCs showed a significant correlation regarding response to induction chemotherapy on day 28. While MDSCs and not MUC1 are associated with inferior response to induction chemotherapy on day 28.ConclusionThe current data suggested that AML patients exhibit an increased presence of MDSCs as well as MUC1 gene overexpression in comparison with normal controls. While MDSCs showed a significant correlation regarding response to induction chemotherapy on day 28, MDSCs and not MUC1 are associated with inferior response to induction chemotherapy on the same day.

Clinical significance of changes of myeloid-derived suppressor cells and their subtypes in inflammatory bowel disease

Objective To explore the changes and their clinical significance of peripheral blood myeloid-derived suppressor cells (MDSC) and their subtypes in patients with inflammatory bowel disease (IBD). Methods From April 2016 to April 2017, 99 hospitalized IBD patients in 2nd Xiangya Hospital of Central South University were enrolled as observation group one, which included 84 Crohn′s disease (CD) (70 in active phase and 14 in remission phase) and 15 patients with ulcerative colitis(UC). At the same period, 32 healthy controls were enrolled as healthy control group one. The proportion of peripheral blood MDSC and subtypes of CD patients, UC patients and healthy controls were examined. Observation group two including 62 IBD patients (47 CD and 15 UC) were selected from observation group one and 21 healthy individuals were selected from healthy control group one as healthy control. The serum levels of tumor necrosis factor α (TNF-α) and interleukin 8 (IL-8) were detected. Chi square test, t test and one-way analysis of variance were performed for statistical analysis. Pearson correlation was performed for correlation analysis. Results The proportion of MDSC in peripheral blood mononuclear cells of CD and UC patients of observation group one were both higher than that of healthy control group one ((6.30±3.97)% and (7.50±3.12)% vs. (3.94±2.25)%, respectively), and the differences were statistically significant (t=-3.22 and -3.21, both P<0.01). The proportion of granulocytic MDSC in peripheral blood mononuclear cells of CD patients was higher than that of UC patients and healthy control group one ((65.69±20.45)% vs. (50.93±13.56)% and (51.50±11.61)%, respectively), and the differences were statistically significant (t=2.93 and 3.79, both P<0.01). The proportion of monocytic MDSC in peripheral blood mononuclear cells of UC patients was higher than that of CD patients and healthy control group one ((28.41±18.33)% vs. (18.38±17.43)% and (28.17±10.22)%, respectively), and the differences were statistically significant (t=2.22 and 2.93, both P<0.05). The proportion of granulocytic MDSC was higher and the proportion of monocytic MDSC was lower in peripheral blood mononuclear cells of CD patients in active phase than those of CD patients in remission phase ((67.36±2.27)% vs. (46.49±6.32)%, and (17.19±2.02)% vs. (34.33±6.12)%), and the differences were statistically significant (t=3.60 and 3.26, both P<0.01). The serum level of TNF-α of CD patients of observation group two was higher than that of UC patients and healthy control group two ((7.83±6.54) ng/L vs. (4.77±2.12) ng/L and (4.40±2.05) ng/L), and the differences were statistically significant (t=2.01 and 2.53, both P<0.05). The serum level of IL-8 of UC patients of observation group two was higher than that of CD patients and healthy control group two ((65.80±45.14) ng/L vs. (25.80±22.32) ng/L and (26.40±22.37) ng/L), and the differences were statistically significant (t=4.87 and 4.21, both P<0.01). Granulocytic MDSC was positively correlated with TNF-α (r=0.319, P=0.011) and was negatively correlated with IL-8 (r=-0.296, P=0.019). Monocytic MDSC was negatively correlated with TNF-α (r=-0.260, P=0.040) and was positively correlated with IL-8 (r=0.306, P=0.016). Conclusions The proportion of granulocytic MDSC in peripheral blood mononuclear cells significantly increases in active CD patients, while the proportion of monocytic MDSC significantly increases in UC and CD patients in remission phase. Detection of MDSC and their subtypes maybe helpful in the differentiation of CD and UC as well as the diagnosis and treatment of CD. Key words: Inflammatory bowel diseases; Tumor necrosis factor α; Interleukin 8; Myeloid-derived suppressor cells and subsets

Abstract 3679: VEGF-A is increased and correlates with MDSC-driven immune suppression, systemic inflammation, nutritional impairment and poor prognosis in patients with cancer

Abstract Vascular Endothelial Growth Factor (VEGF) is a key factor for tumor progression through induction of angiogenesis and other actions including immune alteration. Myeloid-derived suppressor cells (MDSC) are a major type of immune-suppressing cell that appear in cancer or inflammation and that have been reported to express the VEGF-receptor and to be activated by VEGF. To study the clinical importance of VEGF and MDSC for cancer, peripheral blood was collected from patients with gastrointestinal, ovarian, breast, thyroid and pulmonary cancers. Peripheral blood mononuclear cells (PBMC) were separated using a Ficoll-density gradient and were used for the detection of MDSC (CD11b+CD14-CD33+) using flow cytometry and for cytokine-production assays. For these assays, PBMC were stimulated with PHA and the production of cytokines including IL-12 (Th1 inducer), IL-10, (anti-inflammatory cytokine) and IL-17, (pro-inflammatory cytokine), were measured over 24 h using ELISAs. Serum concentrations of IL-10 and VEGF were also measured using ELISAs. The serum levels of both VEGF and MDSC were significantly increased in almost all types of cancer tested and significantly correlated with each other. Their levels also significantly correlated with neutrophil/lymphocyte ratios (NLR) (inflammation marker) and CRP levels, and were inversely correlated with the PHA-stimulation index (SI) (cell mediated immune responses marker) and serum concentrations of rapid-turnover protein (RTP) (nutrition marker). VEGF levels also correlated with serum concentrations of IL-10 and VEGF, and production of IL-17, and inversely correlated with production of IL-12. The prognosis of stage IV colorectal cancer with high VEGF was significantly worse than that with low VEGF. In thyroid cancer, the number of MDSC was significantly higher, NLR and CRP levels were higher, and RTP levels were lower in patients with undifferentiated carcinoma than in those with differentiated carcinoma including papillary and follicular carcinomas. Thus VEGF was increased in cancer and correlated with immune suppression driven by MDSC, inflammation and malnutrition. Although cancer immunotherapy is currently in use for a number of cancers, MDSC have been reported to be a major inhibitor of cancer immunotherapy even in cases in which an immune checkpoint inhibitor was used. An anti-VEGF treatment strategy has now been established in combination with chemotherapy for many types of cancer. Among various types of anti-MDSC trials, anti-VEGF treatment seems to be an effective adjuvant therapy of cancer immunotherapy. Citation Format: Masahiko Shibata, Kenji Gonda, Takahiro Nakajima, Koji Kono, Hiroyuki Suzuki, Seiichi Takenoshita. VEGF-A is increased and correlates with MDSC-driven immune suppression, systemic inflammation, nutritional impairment and poor prognosis in patients with cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3679. doi:10.1158/1538-7445.AM2017-3679

Pattern of glycosylphosphatidylinositol-anchored 80 kD protein (GPI-80) expression in peripheral blood as a marker for myeloid-derived suppressor cells (MDSCs) in metastatic renal cell cancer (mRCC) patients.

504 Background: Metastatic renal cell cancer (mRCC) patients are afflicted with the immunosuppressed condition, and myeloid-derived suppressor cells (MDSCs) are considered to be associated with this condition. MDSCs are difficult to detect in peripheral blood as their cell structures are not uniform, resembling both neutrophils and monocytes. GPI-80 is known as a regulator of Mac-1 (CD11b/CD18) and closely related with neutrophilic maturation. In this study, we investigated whether the pattern of GPI-80 expression could serve as a useful marker for the detection of MDSCs in peripheral blood among mRCC patients. Methods: This study was approved by the Ethics Committee of Yamagata University’s Faculty of Medicine. Peripheral blood samples, collected from 12 mRCC patients before treatment and 12 healthy volunteers, were stained with several differentiation markers such as CD11b, CD16, CD15, CD33, and GPI-80. The samples were examined using flow cytometry to detect unusual differentiation of myeloid cells. To investigate the immunosuppressive function of GPI-80 positive cells, reactive oxygen species (ROS) was measured, and T cell proliferation assay was performed. Results: The coefficient of variation (CV) of GPI-80 was significantly augmented in the CD16hi neutrophil cell population (p = 0.0001). The mean fluorescence intensity (MFI) of GPI-80 was significantly increased in the CD33hi monocyte cell population (p = 0.0074) in the peripheral blood cells of mRCC patients. Furthermore, the GPI-80 CV in the CD16hi population was inversely correlated with the proliferative ability of T cells (p = 0.0214), and the GPI-80 MFI of the CD33hi population was correlated with ROS production (p = 0.0497). Conclusions: These results showed that the pattern of GPI-80 expression in peripheral blood cells is a simple and useful marker for unusual differentiation, which is linked to MDSC functions among mRCC patients. Further studies are, however, required to identify the role of GPI-80 as a prognostic or predictive marker in such patients.

Circulating myeloid-derived suppressor cells (MDSC) and correlation to poor prognosis, Th2-polarization, inflammation, and nutritional damages in patients with gastric cancer.

3063 Background: Recent studies have shown that myeloid-derived suppressor cells (MDSC), which have been identified in most patients, are potent suppressors of T cell activation. MDSC have been reported to be correlated with tumorigenesis and the progression of tumors. We have previously reported significant correlations of MDSC with nutritional damage and inflammation in many types of cancer. In this study the quantitative detection of MDSC in peripheral blood was analyzed in the prognosis in patients with gastric cancer. Methods: We tested PBMCs from 29 preoperative patients with gastric cancer and 18 healthy volunteers using flow cytometric analysis (CD14-CD11b+CD33+). Markers for nutritional status (serum levels of total protein), inflammation (NLR: neurphil/lymphocyte ratio, serum levels of vascular endothelial growth factor: VEGF) and Th2-polarization (PBMC’s production of IL-6 and IL-10) were measured in this study. The prognosis of the patients was assessed by Kaplan-Meier tests for correlation with levels of MDSC. Results: MDSC levels in 29 preoperative patients with gastric cancer were significantly higher than in normal volunteers. These levels were not associated with pathological characteristics. However they were significantly correlated with the production of IL-6 and IL-10, VEGF levels and NLR. A significant inverse correlation with levels of total protein was obtained. MDSCs were increased in stage IV patients compared with healthy volunteer and 2-year survival rate of the patients with higher levels of MDSC (&gt; 1.104%PBMC) was significantly poorer (median OS, 498 vs 473 days; p = 0.048), while there was no significant difference in patients with stages I, II, and III. Conclusions: These data suggest that increased MDSC is an effective biomarker for immunosuppression due to Th2-polarization, nutritional impairment, systemic inflammation and poor prognosis in advanced patients with gastric cancer.

Abstract 5416: Myeloid-derived suppressor cells (MDSC) are increased and correlated with immune suppression and chronic inflammation in patients with cancer

Abstract Chronic inflammation plays a key role in the progress of malignant diseases and has been reported to relate with immune suppression seen in patients with advanced diseases. MDSC: myeloid-derived suppressor cells found as a new type of immune suppressor cells are enhanced by inflammation and have been reported to be present in blood circulation, lymphnodes and tumor tissues in patients with cancer. The suppression of immune function via depleting arginine and producing immune-suppressing cytokines including IL-10 has also been reported. In order to investigate MDSC in patients, peripheral blood mononuclear cells (PBMC) were collected from 18 normal healthy volunteers, 53 patients with gastrointestinal cancer, 65 with breast cancer, 8 with thyroid cancer, and 5 with ovarian cancer. These cells were used for the detection of MDSC (CD11b+CD14−CD33+) by flow cytometry. PBMC was also used for the PHA-blastogenesis of lymphocytes which is a marker of cell mediated immunity (stimulation indices: SI) and for the production assay of cytokines including IL-6 and IL-10. MDSC (%PBMC) of whole patients (5.68 + 5.92%), those of gastrointestinal cancer patients (3.76 + 4.90%) and those of breast cancer patients (2.92 + 4.13%) were significantly higher than those of normal volunteers (p&amp;lt;0.005, p=0.05, p&amp;lt;0.05, respectively, vs 1.59 + 1.08%). MDSC were inversely correlated with the SI (r=−0.271, p&amp;lt;0.05) in patients with gastrointestinal cancer and positively correlated with IL-6 production in whole patients(r=0.287, p=0.05) and with IL-10 production in patients with breast cancer (r=0.363, p&amp;lt;0.05). They were also significantly correlated to neutrophil counts (r=0.287, p&amp;lt;0.05) and neutrophil/lymphocyte ratios (NLR, r=0.623, p=0.05), and inversely correlated to lymphocyte counts (r=−0.251, p&amp;lt;0.01). Thus MDSC were successfully detected in peripheral blood and were significantly higher in patients with cancer. There were strong relationships among the inhibition of cell-mediated immunity, production of type 2 cytokines and inflammatory markers such as NLR. Therefore, MDSC seemed to work as a suppressor of immune reaction and may be an important marker of chronic inflammation in patients with cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5416. doi:1538-7445.AM2012-5416

Relationship of myeloid-derived suppressor cells (MDSC) and immune suppression, nutritional impairment, and inflammatory markers in patients with gastrointestinal cancer.

675 Background: A suppression of cell mediated immunity and malnutrition are commonly seen in patients with advanced cancer and it has been reported that chronic inflammation plays a key role in induction of these conditions. MDSC: myeloid-derived suppressor cells, found as a new type of immune suppressor cells that is closely related to chronic inflammation, have reported to be present in blood circulation in patients with cancer. MDSC have been reported to suppress immune function through several pathways. Methods: Peripheral blood mononuclear cells (PBMC) were collected from 18 normal healthy volunteers, and 53 patients with gastrointestinal cancer including 5 patients with esophageal, 16 with gastric, 26 with colorectal, 3 with bile duct, 1 with pancreatic and 1 with hepatocellular carcinomas, and these cells were used for the detection of MDSC (CD11b+CD14-CD33+) by flow cytometry. PBMC was also used for the PHA-blastogenesis of lymphocytes which is a marker of cell mediated immunity (stimulation indices: SI). Results: MDSC(%PBMC) of whole patients with gastrointestinal cancer, those with gastric cancer and those with colorectal cancer were higher than those of healthy volunteer (3.76+4.90%, 5.18+1.86%, 4.21+1.43% vs 1.59+1.08%, p&lt;0.05, p&lt;0.05, p&lt;0.10, respectively). MDSC of patients with gastrointestinal cancer were also inversely correlated to the SI (r=−0.271, p&lt;0.05) and to the serum concentrations of total protein (r=−0.490, p&lt;0.005). It also tended to correlate to neutrophil counts (r=0.287, p&lt;0.10) and neutrophil/lymphocyte ratios (NLR, r=0.623, p=0.10), and inversely did to lymphocyte counts (r=−0.251, p&lt;0.10).Thus MDSC was successfully detected in peripheral blood and was significantly higher in patients with gastrointestinal cancer. It was related to the inhibition of cell-mediated immunity, hypoaproteinemia and inflammatory markers such as NLR. Conclusions: MDSC seemed to work in the mechanisms of suppression of immune reaction and malnutrition, typically seen in cancer cachexia, in patients with gastrointestinal cancer and may be important as a marker of advancement of malignant diseases.