Additional file 3: of Granulocyte macrophage colony-stimulating factor receptor Îą expression and its targeting in antigen-induced arthritis and inflammation

Genes significantly changed in CD115+ cells from day 4 AIP following CAM-3003, CAT-004 or PBS treatment (day -1). CD115+ PECs were sorted from the peritoneal cavity of C57BL/6 on day 4 and subjected to microarray analysis. Highlighted genes were increased in CAM-3003 vs. CAT-004-treated mice; all other genes were decreased in CAM-3003- vs. CAT-004- or PBS-treated mice. (PDF 25 kb

T‐cell responses and therapies against SARS‐CoV‐2 infection

Coronavirus disease 2019 (COVID-19) is caused by SARS-CoV-2, a novel coronavirus strain. Some studies suggest that COVID-19 could be an immune-related disease, and failure of effective immune responses in initial stages of viral infection could contribute to systemic inflammation and tissue damage, leading to worse disease outcomes. T cells can act as a double-edge sword with both pro- and anti-roles in the progression of COVID-19. Thus, better understanding of their roles in immune responses to SARS-CoV-2 infection is crucial. T cells primarily react to the spike protein on the coronavirus to initiate antiviral immunity; however, T-cell responses can be suboptimal, impaired or excessive in severe COVID-19 patients. This review focuses on the multifaceted roles of T cells in COVID-19 pathogenesis and rationalizes their significance in eliciting appropriate antiviral immune responses in COVID-19 patients and unexposed individuals. In addition, we summarize the potential therapeutic approaches related to T cells to treat COVID-19 patients. These include adoptive T-cell therapies, vaccines activating T-cell responses, recombinant cytokines, Th1 activators and Th17 blockers, and potential utilization of immune checkpoint inhibitors alone or in combination with anti-inflammatory drugs to improve antiviral T-cell responses against SARS-CoV-2. Other Information Published in: Immunology License: http://creativecommons.org/licenses/by/4.0/See article on publisher's website: http://dx.doi.org/10.1111/imm.13262</p

Transcriptomic Profiling of Circulating HLA-DR^– Myeloid Cells, Compared with HLA-DR⁺ Myeloid Antigen-presenting Cells

Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of cells with potent immunosuppressive functions, which can inhibit the activation of immune responses under a steady-state condition and pathological conditions. We performed transcriptomic profiling of circulating CD33+HLA-DR+ myeloid antigen-presenting cells (APCs) and CD33+HLA-DR– myeloid cells (potentially MDSCs) in healthy individuals. We sorted both subpopulations from peripheral blood mononuclear cells (PBMCs) of 10 healthy donors and performed RNA sequencing (RNA-Seq). We found that several signaling pathways associated with the positive regulation of immune responses, such as antigen presentation/processing, FcγR-mediated phagocytosis and immune cell trafficking, phosphoinositide 3-kinase (PI3K)/Akt signaling, DC maturation, triggering receptor expressed on myeloid cells 1 (TREM1) signaling, nuclear factor of activated T cells (NFAT) and IL-8 signaling were downregulated in CD33+HLA-DR– myeloid cells. In contrast, pathways implicated in tumor suppression and anti-inflammation, including peroxisome proliferator-activated receptor (PPAR) and phosphatase and tensin homolog (PTEN), were upregulated in CD33+HLA-DR– myeloid cells. These data indicate that PPAR/PTEN axis could be upregulated in myeloid cells to keep the immune system in check in normal physiological conditions. Our data reveal some of the molecular and functional differences between CD33+HLA-DR+ APCs and CD33+HLA-DR– myeloid cells in a steady-state condition, reflecting the potential suppressive function of CD33+HLA-DR– myeloid cells to maintain immune tolerance. For future studies, the same methodological approach could be applied to perform transcriptomic profiling of myeloid subsets in pathological conditions.Other InformationPublished in: Immunological InvestigationsLicense: http://creativecommons.org/licenses/by-nc-nd/4.0/See article on publisher's website: https://dx.doi.org/10.1080/08820139.2020.1795875</p

Additional file 1: of CCL17 blockade as a therapy for osteoarthritis pain and disease

Synovial cell populations from joints at week 1 from WT, Irf4 −/− , Ccl17 E/E and GM-CSF −/− mice undergoing CiOA were sorted and gene expression measured. (A) Representative FACS plots showing synovial cell sorting strategy. CD45+ cells (II) were sorted into neutrophils (CD11b+Ly6G+) (III) and macrophages (CD11b+Ly6G−F4/80+CD64+) (IV); CD45− cells (I) were sorted into endothelial cells (CD31+mEF-SK4−) (V), fibroblasts (CD31−mEF-SK4+) (VII) and other cells (CD31−mEF-SK4−) (VI). (B-D) mRNA expression in sorted synovial cell populations. (B) Col1a1, (C) Mmp3 and (D) Mmp13. Results are expressed as the mean ± SEM; n = 3–6 mice per strain. N.D. not detected. *p < 0.05, **p < 0.01, WT vs. Irf4−/−, Ccl17 E/E or GM-CSF−/−mice. (PDF 257 kb

Additional file 2: of CCL17 blockade as a therapy for osteoarthritis pain and disease

Proportions of synovial cell populations in knee joints from CiOA mice at day 7. (PDF 85 kb

Differential gene expression of tumor-infiltrating CD33+ myeloid cells in advanced- versus early-stage colorectal cancer

Colorectal cancer (CRC) has high mortality rates, especially in patients with advanced disease stages, who often do not respond to therapy. The cellular components of the tumor microenvironment are essentially responsible for dictating disease progression and response to therapy. Expansion of different myeloid cell subsets in CRC tumors has been reported previously. However, tumor-infiltrating myeloid cells have both pro- and anti-tumor roles in disease progression. In this study, we performed transcriptomic profiling of cells of myeloid lineage (CD33+) from bulk CRC tumors at varying disease stages. We identified differentially expressed genes and pathways between CRC patients with advanced stage and early stages. We found that pro-angiogenic and hypoxia-related genes were upregulated, while genes related to immune and inflammatory responses were downregulated in CD33+ myeloid cells from patients with advanced stages, implying that immune cell recruitment and activation could be compromised in advanced disease stages. Moreover, we identified a unique “poor prognosis CD33+ gene signature” by aligning top upregulated and downregulated genes in tumor-infiltrating myeloid cells from our analyses with data from The Cancer Genome Atlas. Our results showed that this gene signature is an independent prognostic indicator for disease-specific survival in CRC patients, potentially reflecting its clinical importance.Other Information Published in: Cancer Immunology, Immunotherapy License: https://creativecommons.org/licenses/by/4.0See article on publisher's website: http://dx.doi.org/10.1007/s00262-020-02727-0</p

MOESM3 of Transcriptomic profiling disclosed the role of DNA methylation and histone modifications in tumor-infiltrating myeloid-derived suppressor cell subsets in colorectal cancer

Additional file 3: Table S2.: DAVID analysi

MOESM2 of Transcriptomic profiling disclosed the role of DNA methylation and histone modifications in tumor-infiltrating myeloid-derived suppressor cell subsets in colorectal cancer

Additional file 2: Table S1. Gene cluster analysis

MOESM1 of Transcriptomic profiling disclosed the role of DNA methylation and histone modifications in tumor-infiltrating myeloid-derived suppressor cell subsets in colorectal cancer

Additional file 1: Figure S1. Adherent junction pathway in tumor-infiltrating PMN-MDSCs. The upregulated genes in PMN-MDSCs from 2 patients were uploaded in DAVID to identify the biological pathways. Adherent junction is the top KEGG pathway regulated in PMN-MDSCs, compared with APCs within the TME. The black ovals highlight the KEGG identified functional pathways that are regulated by PMN-MDSCs, within the TME. The functional consequences of related genes are shown in red. Figure S2. HIF-1 signaling pathway in tumor-infiltrating I-MDSCs. The downregulated genes in PMN-MDSCs, compared with I-MDSCs from two patients were uploaded in DAVID to identify the biological pathways. HIF-1 signaling is the top KEGG pathway regulated in I-MDSCs, within the TME. The black ovals highlight the KEGG identified functional pathways that are regulated by I-MDSCs, within the TME. The functional consequences of related genes are shown in red. Figure S3. Validation of differential gene expression and functional network analyses of APCs, PMN-MDSCs and I-MDSCs in CRC patients. Heat maps show the TPM representing fold change to the mean expression of WNT signaling, SNARE signaling and JNK pathway activation in I-MDSCs (A). Heat maps show the TPM representing fold change relative to the mean expression of colorectal cancer-, cell migration-, NFκB-, IL-1β production-related genes in PMN-MDSCs, compared with APCs (B). Heat map shows the TPM representing fold change relative to the mean expression of tumor progression-, migration and metastasis- and DNA methylation-related genes in PMN-MDSCs (C). Results obtained from four CRC patients (#09, #12, #13, and #16). Figure S4. CD11a expression in tumor-infiltrating PMN-MDSCs. Heat map shows the TPM representing fold change relative to the mean expression of CD11a gene (ITGAL) in PMN-MDSCs (A). Cells isolated from TT of #07 and #08 patients were stained for myeloid cell markers and CD11a, and analyzed by flow cytometry. Representative flow cytometric plots show the gating strategy employed to identify I-MDSCs and PMN-MDSCs expressing CD11a (B)

Expression of immune checkpoints and T cell exhaustion markers in early and advanced stages of colorectal cancer

Despite recent advances in colorectal cancer (CRC) treatment, a large proportion of patients show limited responses to therapies, especially in advanced stages. There is an urgent need to identify prognostic biomarkers and/or therapeutic targets in advanced stages, aiming to improve the efficacy of current treatments. We aimed to determine prognostic biomarkers in tumor tissue and circulation of CRC patients, with a special focus on T cell exhaustion markers. We found that mRNA levels of PD-1, TIM-3, CTLA-4, TIGIT, CD160, CD244, KLRG1, TOX2, TOX3, Ki-67, and PRDM1 were elevated in CRC tumor tissues. We also investigated differences in gene expression between early and advanced disease stages. We found that TOX and potentially TIM-3, CTLA-4, VISTA, TIGIT, KLRG1, TOX2, SIRT1, Ki-67, and Helios mRNA levels in tumor tissue were elevated in advanced disease stages, suggesting their potential roles in CRC progression. In contrast, PD-1 and CD160 levels in tumor tissue were downregulated in advanced stages. In the circulation of CRC patients, mRNA levels of PD-1, VISTA and LAG-3 were higher than those of healthy individuals. Moreover, in circulation, PD-1, CTLA-4 and TIGIT mRNA levels were reduced in advanced stages. Interestingly, levels of PD-1 in both tumor tissue and circulation were reduced in advanced stages, suggesting that targeting PD-1 in patients with advanced stages could be less effective. Altogether, these findings suggest some potential T cell exhaustion markers that could be utilized as prognostic biomarkers and/or therapeutic targets for CRC. However, further investigations and validations in larger cohorts are required to confirm these findings.Other Information Published in: Cancer Immunology, Immunotherapy License: https://creativecommons.org/licenses/by/4.0See article on publisher's website: http://dx.doi.org/10.1007/s00262-020-02593-w</p