Inflammation and macrophage polarization in cutaneous melanoma: Histopathological and immunohistochemical study

Tumor-associated macrophages (TAMs) are considered to affect tumor growth and progression. Macrophages can be classified into two states of polarized activation, namely classically activated M1 macrophages and alternatively activated M2 macrophages. The dynamic balance between TAMs and tumor cells has an important impact on tumor homeostasis and progression. The aim of this study was to characterize the phenotype of TAMs present in different subtypes of superficial spreading cutaneous melanoma and their relationship with the lymphocytic infiltrate in order to identify new histopathological tools for melanoma prognosis and suitable targets for melanoma therapy. We selected four groups of patients with malignant melanoma in order to analyze the profile of polarized macrophage activation using immunohistochemical methods. Histopathological analysis showed that the macrophage polarization state appears to be more related to the lymphocytic infiltrate than to the thickness of the lesions. Further studies are necessary to increase understanding of the immunopathological dynamic of melanoma that may be modulated by future targeted immunotherapies.

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Macrophage Polarization and Osteoporosis: A Review

Nutrients ◽

10.3390/nu12102999 ◽

2020 ◽

Vol 12 (10) ◽

pp. 2999

Author(s):

Joseph Muñoz ◽

Neda S. Akhavan ◽

Amy P. Mullins ◽

Bahram H. Arjmandi

Keyword(s):

Bone Resorption ◽

Bone Mineralization ◽

Macrophage Polarization ◽

Polarization State ◽

M2 Macrophages ◽

Cytokines And Chemokines ◽

Polarization States ◽

Classically Activated Macrophages ◽

Cost Efficient ◽

Alternatively Activated

Over 200 million people suffer from osteoporosis worldwide. Individuals with osteoporosis have increased rates of bone resorption while simultaneously having impaired osteogenesis. Most current treatments for osteoporosis focus on anti-resorptive methods to prevent further bone loss. However, it is important to identify safe and cost-efficient treatments that not only inhibit bone resorption, but also stimulate anabolic mechanisms to upregulate osteogenesis. Recent data suggest that macrophage polarization may contribute to osteoblast differentiation and increased osteogenesis as well as bone mineralization. Macrophages exist in two major polarization states, classically activated macrophages (M1) and alternatively activated macrophage (M2) macrophages. The polarization state of macrophages is dependent on molecules in the microenvironment including several cytokines and chemokines. Mechanistically, M2 macrophages secrete osteogenic factors that stimulate the differentiation and activation of pre-osteoblastic cells, such as mesenchymal stem cells (MSC’s), and subsequently increase bone mineralization. In this review, we cover the mechanisms by which M2 macrophages contribute to osteogenesis and postulate the hypothesis that regulating macrophage polarization states may be a potential treatment for the treatment of osteoporosis.

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Changes in alpha-7 nicotinic acetylcholine receptor and macrophage polarization state participate in the regulation of cervical remodeling in pregnant rats†

Biology of Reproduction ◽

10.1093/biolre/ioz133 ◽

2019 ◽

Author(s):

Jinying Yang ◽

Yumian Lai ◽

Juanhua Chen ◽

Baohua Lin ◽

Bei Zhou ◽

...

Keyword(s):

Western Blotting ◽

Macrophage Polarization ◽

Polarization State ◽

M2 Macrophages ◽

Pregnant Rats ◽

M1 Macrophages ◽

Nicotinic Acetylcholine ◽

Immunohistochemistry Staining ◽

Cervical Remodeling ◽

Alpha 7

Abstract To test the hypothesis that changes in alpha-7 nicotinic acetylcholine receptor (α7nAChR) expression on macrophages and macrophage polarization participate in cervical remodeling during normal pregnancy, pregnant rats from gestational days (GDs) 14, 16, 18, 20, and 22 were used in the present study. The expression of α7nAChR on macrophages and the numbers of M1 and M2 macrophages were detected by double immunofluorescence staining. The levels of α7nAChR and collagens were detected by western blotting. M1 markers (inducible nitric oxide synthase and inflammatory cytokines) and M2 markers (arginase 1, anti-inflammatory cytokines) were detected to evaluate the macrophage polarization state by immunohistochemistry staining, western blotting, and the enzyme-linked immunosorbent assay. Matrix metalloproteinase 9 (MMP-9) expression was determined by immunohistochemistry staining and western blotting. We found that the α7nAChR expression on macrophages significantly decreased on GD22 compared to GDs 14, 16, 18, and 20. There was an increased number of M1 macrophages and decreased number of M2 macrophages in late pregnancy. The expression of M1 macrophage biomarkers was much higher on GDs 20 and 22 than on GDs 14, 16, and 18, but expression of M2 biomarkers decreased on GDs 20 and 22 compared to GDs 14, 16, and 18. MMP-9 expression was higher on GD22 than on GDs 14, 16, 18, and 20, and collagen expression significantly decreased on GDs 18, 20, and 22 compared to GD14. Our results indicated that the decreased expression of α7nAChR and increased number of M1 macrophages are associated with cervical remodeling.

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Macrophage Polarization in Cardiac Tissue Repair Following Myocardial Infarction

International Journal of Molecular Sciences ◽

10.3390/ijms22052715 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2715

Author(s):

Yevgeniy Kim ◽

Sanzhar Nurakhayev ◽

Ayan Nurkesh ◽

Zharylkasyn Zharkinbekov ◽

Arman Saparov

Keyword(s):

Myocardial Infarction ◽

Cardiovascular Disease ◽

Cardiac Tissue ◽

Macrophage Polarization ◽

Scar Tissue ◽

M1 Macrophages ◽

Mortality And Morbidity ◽

Phenotypic Profile ◽

Detrimental Impact ◽

Alternatively Activated

Cardiovascular disease is the leading cause of mortality and morbidity around the globe, creating a substantial socio-economic burden as a result. Myocardial infarction is a significant contributor to the detrimental impact of cardiovascular disease. The death of cardiomyocytes following myocardial infarction causes an immune response which leads to further destruction of tissue, and subsequently, results in the formation of non-contractile scar tissue. Macrophages have been recognized as important regulators and participants of inflammation and fibrosis following myocardial infarction. Macrophages are generally classified into two distinct groups, namely, classically activated, or M1 macrophages, and alternatively activated, or M2 macrophages. The phenotypic profile of cardiac macrophages, however, is much more diverse and should not be reduced to these two subsets. In this review, we describe the phenotypes and functions of macrophages which are present in the healthy, as well as the infarcted heart, and analyze them with respect to M1 and M2 polarization states. Furthermore, we discuss therapeutic strategies which utilize macrophage polarization towards an anti-inflammatory or reparative phenotype for the treatment of myocardial infarction.

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Dynamic macrophage polarization-specific miRNA patterns reveal increased soluble VEGF receptor 1 by miR-125a-5p inhibition

Physiological Genomics ◽

10.1152/physiolgenomics.00098.2015 ◽

2016 ◽

Vol 48 (5) ◽

pp. 345-360 ◽

Cited By ~ 10

Author(s):

David W. Melton ◽

XiuFen Lei ◽

Jonathan A. L. Gelfond ◽

Paula K. Shireman

Keyword(s):

Culture Media ◽

Macrophage Polarization ◽

Expression Patterns ◽

Polarization State ◽

Multiple Time ◽

Vegf Receptor ◽

M1 Macrophages ◽

M1 Macrophage ◽

Macrophage Phenotypes ◽

Multiple Time Points

Dynamic, epigenetic mechanisms can regulate macrophage phenotypes following exposure to different stimulating conditions and environments. However, temporal patterns of microRNAs (miRNAs or miRs) across multiple macrophage polarization phenotypes have not been defined. We determined miRNA expression in bone marrow-derived murine macrophages over multiple time points (0.5, 1, 3, 24 h) following exposure to cytokines and/or LPS. We hypothesized that dynamic changes in miRNAs regulate macrophage phenotypes. Changes in macrophage polarization markers were detected as early as 0.5 and as late as 24 h; however, robust responses for most markers occurred within 3 h. In parallel, many polarization-specific miRNAs were also changed by 3 h and expressed divergent patterns between M1 and M2a conditions, with increased expression in M1 (miR-155, 199a-3p, 214-3p, 455-3p, and 125a) or M2a (miR-511 and 449a). Specifically, miR-125a-5p exhibited divergent patterns: increased at 12–24 h in M1 macrophages and decreasing trend in M2a. VEGF in the culture media of macrophages was dependent upon the polarization state, with greatly diminished VEGF in M2a compared with M1 macrophage culture media despite similar VEGF in cell lysates. Inhibition of miR-125a-5p in media-only controls (MO) and M1 macrophages greatly increased expression and secretion of soluble VEGF receptor-1 (sVEGFR1) leading to diminished VEGF in the culture media, partially converting MO and M1 into an M2a phenotype. Thus, the divergent expression patterns of polarization-specific miRNAs led to the identification and demonstrated the regulation of a specific macrophage polarization phenotype, sVEGFR1 by inhibition of miR-125a-5p.

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Defects in Macrophage Reprogramming in Cancer Therapy: The Negative Impact of PD-L1/PD-1

Frontiers in Immunology ◽

10.3389/fimmu.2021.690869 ◽

2021 ◽

Vol 12 ◽

Author(s):

Hao Cai ◽

Yichi Zhang ◽

Jian Wang ◽

Jinyang Gu

Keyword(s):

Cancer Therapy ◽

Negative Impact ◽

Macrophage Polarization ◽

M2 Macrophage ◽

M2 Macrophages ◽

M1 Macrophages ◽

Programmed Death ◽

Antigen Presenting ◽

Cell Death Protein ◽

Alternatively Activated

Classically activated M1 macrophages and alternatively activated M2 macrophages are two polarized subsets of macrophages at the extreme ends of a constructed continuum. In the field of cancer research, M2 macrophage reprogramming is defined as the repolarization of pro-tumoral M2 to anti-tumoral M1 macrophages. It is known that colony-stimulating factor 1 (CSF1)/CSF1 receptor (CSF1R) and CSF2/CSF2R signaling play important roles in macrophage polarization. Targeting CSF1/CSF1R for M2 macrophage reprogramming has been widely performed in clinical trials for cancer therapy. Other targets for M2 macrophage reprogramming include Toll-like receptor 7 (TLR7), TLR8, TLR9, CD40, histone deacetylase (HDAC), and PI3Kγ. Although macrophages are involved in innate and adaptive immune responses, M1 macrophages are less effective at phagocytosis and antigen presenting, which are required properties for the activation of T cells and eradication of cancer cells. Similar to T and dendritic cells, the “functionally exhausted” status might be attributed to the high expression of programmed death-ligand 1 (PD-L1) or programmed cell death protein 1 (PD-1). PD-L1 is expressed on both M1 and M2 macrophages. Macrophage reprogramming from M2 to M1 might increase the expression of PD-L1, which can be transcriptionally activated by STAT3. Macrophage reprogramming or PD-L1/PD-1 blockade alone is less effective in the treatment of most cancers. Since PD-L1/PD-1 blockade could make up for the defect in macrophage reprogramming, the combination of macrophage reprogramming and PD-L1/PD-1 blockade might be a novel treatment strategy for cancer therapy.

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Iron Released after Cryo-Thermal Therapy Induced M1 Macrophage Polarization, Promoting the Differentiation of CD4+ T Cells into CTLs

International Journal of Molecular Sciences ◽

10.3390/ijms22137010 ◽

2021 ◽

Vol 22 (13) ◽

pp. 7010

Author(s):

Shicheng Wang ◽

Man Cheng ◽

Peng Peng ◽

Yue Lou ◽

Aili Zhang ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Antitumor Immunity ◽

Macrophage Polarization ◽

Thermal Therapy ◽

High Plasticity ◽

Antitumor Effects ◽

M1 Macrophages ◽

Splenic Macrophages ◽

M1 Macrophage

Macrophages play critical roles in both innate and adaptive immunity and are known for their high plasticity in response to various external signals. Macrophages are involved in regulating systematic iron homeostasis and they sequester iron by phagocytotic activity, which triggers M1 macrophage polarization and typically exerts antitumor effects. We previously developed a novel cryo-thermal therapy that can induce the mass release of tumor antigens and damage-associated molecular patterns (DAMPs), promoting M1 macrophage polarization. However, that study did not examine whether iron released after cryo-thermal therapy induced M1 macrophage polarization; this question still needed to be addressed. We hypothesized that cryo-thermal therapy would cause the release of a large quantity of iron to augment M1 macrophage polarization due to the disruption of tumor cells and blood vessels, which would further enhance antitumor immunity. In this study, we investigated iron released in primary tumors, the level of iron in splenic macrophages after cryo-thermal therapy and the effect of iron on macrophage polarization and CD4+ T cell differentiation in metastatic 4T1 murine mammary carcinoma. We found that a large amount of iron was released after cryo-thermal therapy and could be taken up by splenic macrophages, which further promoted M1 macrophage polarization by inhibiting ERK phosphorylation. Moreover, iron promoted DC maturation, which was possibly mediated by iron-induced M1 macrophages. In addition, iron-induced M1 macrophages and mature DCs promoted the differentiation of CD4+ T cells into the CD4 cytolytic T lymphocytes (CTL) subset and inhibited differentiation into Th2 and Th17 cells. This study explains the role of iron in cryo-thermal therapy-induced antitumor immunity from a new perspective.

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POS0333 ALTERED MACROPHAGE POLARIZATION PHENOTYPES IN SYSTEMIC SCLEROSIS

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.3124 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 394.1-394

Author(s):

A. Hukara ◽

M. Rudnik ◽

C. B. Rufer ◽

O. Distler ◽

P. Blyszczuk ◽

...

Keyword(s):

Systemic Sclerosis ◽

Macrophage Polarization ◽

Peritoneal Macrophages ◽

Surface Marker ◽

Surface Marker Expression ◽

Tnf Α ◽

Ifn Γ ◽

Alternatively Activated ◽

Classically Activated

Background:Fos-like 2 (Fosl-2) is a transcription factor of the AP-1 family and has a broad range in inducing cellular changes affecting fibrosis and inflammatory responses. Pathological effects of Fosl-2 have been associated with systemic sclerosis (SSc). In addition, increased expression of Fosl-2 has been detected in human SSc monocyte-derived macrophages [1]. Monocytes and macrophages play a central role in activating and propagating acute inflammation followed by pathological fibrosis and organ dysfunction. The classification of the macrophage polarization phenotype can be assigned based on the stimulus, for example into classically-activated M(LPS), and alternatively-activated M(IL-4) macrophages [2]. However, the role of the Fosl-2 transcription factor in macrophage polarization remains elusive.Objectives:To investigate the role of Fosl-2 in macrophage polarization in SSc using Fosl-2 overexpressing transgenic (Fosl-2 tg) mice and human blood-derived macrophages from SSc patients.Methods:Thiogylcolate-elicited peritoneal macrophages were isolated from wild-type (wt) and Fosl-2 tg mice. Human peripheral CD14+ blood-derived monocytes were isolated and differentiated to macrophages (hMDM) from healthy controls and SSc patients. Murine and human macrophages were polarized with LPS (10 ng/ml), LPS + recombinant mouse IFN-γ (10 ng/ml), recombinant mouse, resp. human IL-4 (10 ng/ml) or remained untreated. Macrophage surface marker expression was assessed by flow cytometry using a mouse (F4/80, CD11b, CD86, CD80, CD38, MHCII, CD206, PD-L1, PD-L2, CD36) or human (CD38, CD40, CD86, PD-L2, PD-L1, CD163, CD206) designed polarization panel. Phagocytic activity was detected with pHrodo Red E.coli particles by flow cytometry. Gene expression and secretion of pro- and anti-inflammatory markers were measured by RT-qPCR, standard ELISAs and Griess Assay for nitric oxide production.Results:After LPS stimulation, mRNA levels of IL-1β (p<0.01, n=11-12), TNF-α (p=0.05, n=11-12) and IFN-γ (p<0.05, n=7) were reduced, whereas expression of IL-10 (p<0.05, n=11-12) was enhanced in Fosl-2 tg peritoneal macrophages in comparison to wt cells. Secretion of TNF-α (p<0.01, n=9-11) and nitric oxide (p<0.01, n=9) was impaired in Fosl-2 tg peritoneal macrophages compared to wt cells after LPS stimulation. Peritoneal macrophages were analyzed directly after isolation for macrophage polarization cell surface marker expression. Fosl-2 tg peritoneal macrophages showed an increase in the F4/80+CD11b+PD-L2+CD36+ cell population (p<0.01, n=3-6) compared to peritoneal macrophages from wt mice.The expression of cell surface markers of non-polarized and IL-4 stimulated SSc hMDM (n=17) showed an increased percentage of CD40+CD86+CD206+PD-L2+CD163+ cells (p<0.05) compared to healthy control hMDM (n=7). Phagocytic activity was enhanced in SSc hMDM (n=7) compared to healthy untreated (p<0.05), LPS (p=0.05) and IL-4 (p<0.05) hMDM (n=5).Conclusion:Our animal data indicates a role of Fosl-2 in regulating macrophage polarization with a shift from a classically-activated to an alternatively-activated phenotype. Similarly, SSc hMDM resemble a functional M(IL-4) alternative macrophage phenotype.Thus, maintaining a balanced proportion of classically- and alternatively-activated macrophage phenotypes may be an effective tool to control macrophage function in SSc.References:[1]Moreno-Moral, A., et al., Changes in macrophage transcriptome associate with systemic sclerosis and mediate GSDMA contribution to disease risk. Ann Rheum Dis, 2018. 77(4): p. 596-601.[2]Kania, G., M. Rudnik, and O. Distler, Involvement of the myeloid cell compartment in fibrogenesis and systemic sclerosis. Nat Rev Rheumatol, 2019. 15(5): p. 288-302.Disclosure of Interests:Amela Hukara: None declared, Michal Rudnik: None declared, Chantal Brigitta Rufer: None declared, Oliver Distler Speakers bureau: Actelion, Bayer, Boehringer Ingelheim, Medscape, Novartis, Roche, Menarini, Mepha, MSD, iQone, Pfizer, Consultant of: Abbvie, Actelion, Acceleron Pharma, Amgen, AnaMar, Arxx Therapeutics, Bayer, Baecon Discovery, Blade Therapeutics, Boehringer, CSL Behring, ChemomAb, Corpuspharma, Curzion Pharmaceuticals, Ergonex, Galapagos NV, GSK, Glenmark Pharmaceuticals, Inventiva, Italfarmaco, iQvia, Kymera, Medac, Medscape, Mitsubishi Tanabe Pharma, MSD, Roche, Sanofi, UCB, Lilly, Target BioScience, Pfizer, Grant/research support from: Actelion, Bayer, Boehringer Ingelheim, Kymera Therapeutics, Mitsubishi Tanabe, Przemyslaw Blyszczuk: None declared, Gabriela Kania: None declared

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Cervus nippon var. mantchuricus water extract treated with digestive enzymes (CE) modulates M1 macrophage polarization through TLR4/MAPK/NF-κB signaling pathways on murine macrophages

European Journal of Inflammation ◽

10.1177/20587392211000898 ◽

2021 ◽

Vol 19 ◽

pp. 205873922110008

Author(s):

Se Hyang Hong ◽

Jin Mo Ku ◽

Ye Seul Lim ◽

Hyo In Kim ◽

Yong Cheol Shin ◽

...

Keyword(s):

Digestive Enzymes ◽

Macrophage Polarization ◽

Water Extract ◽

Cervus Nippon ◽

No Production ◽

M1 Macrophages ◽

Murine Macrophages ◽

M1 Macrophage ◽

Tnf Α ◽

Cytokine Levels

The objective of this study was to investigate the effects of Cervus nippon var. mantchuricus water extract treated with digestive enzymes (CE) on the promotion of M1 macrophage polarization in murine macrophages. Macrophages polarize either to one phenotype after stimulation with LPS or IFN-γ or to an alternatively activated phenotype that is induced by IL-4 or IL-13. Cell viability of RAW264.7 cells was determined by WST-1 assay. NO production was measured by Griess assay. IL-6, IL-12, TNF-α, and iNOS mRNA levels were measured by RT-PCR. IL-6, IL-12, and IL-10 cytokine levels were determined by ELISA. TLR4/MAPK/NF-κB signaling in RAW264.7 cells was evaluated by western blotting. The level of NF-κB was determined by immunoblotting. CE induced the differentiation of M1 macrophages. CE promoted M1 macrophages to elevate NO production and cytokine levels. CE-stimulated M1 macrophages had enhanced IL-6, IL-12, and TNF-α. CE promoted M1 macrophages to activate TLR4/MAPK/NF-κB phosphorylation. M2 markers were downregulated, while M1 markers were upregulated in murine macrophages by CE. Consequently, CE has immunomodulatory activity and can be used to promote M1 macrophage polarization through the TLR4/MAPK/NF-κB signaling pathways.

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