Regulation of the maturation of human monocytes into immunosuppressive macrophages

Bayik, Defne; Tross, Debra; Haile, Lydia; Verthelyi, Daniela; Klinman, Dennis M.

doi:10.1182/bloodadvances.2017011221

Cited by 24 publications

(30 citation statements)

References 56 publications

(82 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Hence, efforts to increase the frequency of immunosuppressive M2-like macrophages represents a potentially useful strategy for the treatment of SLE. Our lab previously showed that the TLR2/1 agonist PAM3 was unique among TLR ligands in promoting the generation of M2-like macrophages from normal human monocytes [14,15]. This work extends those findings to show that PAM3 similarly induces monocytes from lupus patients to preferentially differentiate into M2-like macrophages.…”

Section: Introductionsupporting

confidence: 76%

“…10 5 purified monocytes/well were seeded into 96-well flat bottom plates (Corning) and stimulated for 5 days at 37°C with 1 μg/ml PAM3CSK4 (PAM3) (Invivogen), 250 ng/ml MCSF (Miltenyi Biotec), 250 ng/ml IFNγ (Miltenyi Biotec) or 1 μl/ml R848 (Invivogen). Previous studies established that these were the optimally stimulatory concentrations of each agent ( [14,15] and data not shown). Cells were maintained in medium consisting of RPMI 1640 (Lonza) supplemented with 2% heat-inactivated fetal calf serum 25 nmol/L HEPES, 1 mmol/L sodium pyruvate, non-essential amino acids (NEAAs) and 0.0035% 2mercaptoethanol (2-ME).…”

Section: Isolation and Differentiation Of Human Monocytesmentioning

confidence: 69%

“…Previous studies showed that normal human monocytes preferentially matured into M1-like macrophages when stimulated by most TLR agonists [8,20]. The one exception to this rule was PAM3 which uniquely triggered human monocytes to mature into immunosuppressive M2-like macrophages [14,15]. Given the excess of inflammatory monocytes detected in the blood of lupus patients coupled with evidence from lupus-prone mice that boosting the frequency of M2 macrophages can reduce disease severity [13,21], experiments were undertaken to determine whether PAM3 could stimulate monocytes from lupus patients to mature into M2-like macrophages.…”

Section: Pam3 Induces Lupus Monocytes To Preferentially Mature Into Pmentioning

confidence: 98%

See 2 more Smart Citations

PAM3 supports the generation of M2-like macrophages from lupus patient monocytes and improves disease outcome in murine lupus

Horuluoglu

Bayik

Kayraklioglu

et al. 2019

Journal of Autoimmunity

Self Cite

View full text Add to dashboard Cite

Systematic Lupus Erythematosus (SLE) is an autoimmune syndrome of unclear etiology. While T and B cell abnormalities contribute to disease pathogenesis, recent work suggests that inflammatory M1-like macrophages also play a role. Previous work showed that the TLR2/1 agonist PAM3CSK4 (PAM3) could stimulate normal human monocytes to preferentially differentiate into immunosuppressive M2-like rather than inflammatory M1like macrophages. This raised the possibility of PAM3 being used to normalize the M1:M2 ratio in SLE. Consistent with that possibility, monocytes from lupus patients differentiated into M2-like macrophages when treated with PAM3 in vitro. Furthermore, lupus-prone NZB x NZW F1 mice responded similarly to weekly PAM3 treatment. Normalization of the M2 macrophage frequency was associated with delayed disease progression, decreased autoantibody and inflammatory cytokine synthesis, reduced proteinuria and prolonged survival in NZB x NZW F1 mice. The ability of PAM3 to bias monocyte differentiation in favor of immunosuppressive macrophages may represent a novel approach to the therapy of SLE. not support inflammatory responses. 'Non-classical' monocytes express a CD14 low CD16 + phenotype and are inflammatory. An 'intermediate' or transitional monocyte population also exists that is CD14 + CD16 + and has both phagocytic and inflammatory properties [7]. In response to stimulation, monocytes differentiate into macrophages with diverse phenotypes and functions. These range from 'classical' pro-inflammatory M1-like macrophages that protect the host from infection to 'alternatively activated' M2like macrophages that are immunosuppressive, support tissue remodeling, and phagocytose apoptotic cells [8-10]. Recent studies suggest that monocytes and macrophages can contribute to the pathogenesis of SLE. Indeed, lupus flares are associated with a relative increase in the frequency of M1 vs M2 macrophages [11]. The frequency of inflammatory monocytes is also elevated in SLE patients and the concentration of serum cytokines that promote macrophage differentiation (such as CSF-1 and MCP-1) correlates with disease severity [12]. Normalizing the M1:M2 ratio by adoptive transfer of M2 macrophages is of benefit

show abstract

Section: Introductionsupporting

confidence: 76%

Section: Isolation and Differentiation Of Human Monocytesmentioning

confidence: 69%

Section: Pam3 Induces Lupus Monocytes To Preferentially Mature Into Pmentioning

confidence: 98%

See 1 more Smart Citation

PAM3 supports the generation of M2-like macrophages from lupus patient monocytes and improves disease outcome in murine lupus

Horuluoglu

Bayik

Kayraklioglu

et al. 2019

Journal of Autoimmunity

Self Cite

View full text Add to dashboard Cite

show abstract

“…PAM3 induces murine monocytes to mature into M2 macrophages over four days in vivo {data not shown, section 4.2}. Of note, BMDM cultured with PAM3 for four days in vitro primarily generate M1 macrophages [30][31][32] .This differs from the effect of PAM3 on peripheral blood human monocytes ex vivo and murine monocytes in vivo, where this TLR agonist preferentially generates M2 rather than M1 macrophages 10,18,19 . Other TLR 2/1 agonists also induce the generation of immunosuppressive macrophages whereas ligands that interact with TLR2/6 heterodimers (such as FSL-1) do not (data not shown).…”

Section: Discussionmentioning

confidence: 99%

“…normal murine and human monocytes to preferentially mature into M2 macrophages 18,19 . As an excess of M1 macrophages can contribute to the pathogenesis of colitis 15 , the possibility that the protective activity of PAM3 was mediated via an increase in the number of M2 macrophages was examined.…”

Section: Effect Of Pam3 On M2 Macrophage Frequency Previous Studies mentioning

confidence: 99%

PAM3 protects against DSS-induced colitis by altering the M2:M1 ratio

Horuluoglu

Kayraklioglu

Tross

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

Inflammation of the gastrointestinal tract contributes to the development of inflammatory bowel disease (IBD). Human IBD is modeled by administering dextran sulfate sodium (DSS) to mice. In humans and mice, inflammatory M1 macrophages contribute to the progression of IBD whereas immunosuppressive M2 macrophages protect against colitis. The TLR2/1 agonist PAM3CSK4 (PAM3) induces human and murine monocytes to differentiate into immunosuppressive M2 macrophages, suggesting that PAM3 might be of benefit in the prevention/treatment of colitis. PAM3 was therefore administered to mice treated with DSS. As hypothesized, the number of M2 macrophages rose and disease severity decreased. The critical role of M2 macrophages in this process was established by transferring purified M2 macrophages from PAM3 treated control donors into DSS recipients and reducing colitis. These findings suggest that PAM3 may represent a novel approach to the treatment of human IBD. Ulcerative colitis and Crohn's disease are chronic inflammatory disorders of the gastrointestinal tract 1. In both types of IBD, activation of the innate rather than adaptive immune system is critical, with macrophages and dendritic cells contributing to the induction of inflammation 2-5. Intestinal macrophages occupy the interface between the host's GI tract and the resident microbiome. These macrophages can contribute to IBD pathogenesis by failing to eliminate inflammation-inducing microbes and/or failing to support the resolution of inflammation that arises via other mechanisms 6. The DSS-induced model of murine colitis is widely used to study human IBD due to its rapidity, simplicity and reproducibility. DSS disrupts the colonic epithelium and facilitates the invasion of intestinal microbes through the mucosa, causing inflammation characterized clinically by weight loss, diarrhea and rectal bleeding 7-9. 'Classical' or pro-inflammatory M1-like macrophages protect the host from infection whereas 'alternatively activated' M2-like macrophages act to suppress inflammation and support tissue remodeling 10-13. Studies suggest that M1 and M2 macrophages have opposing roles in DSS-induced colitis 14. M1 macrophages contribute to disease pathogenesis by secreting pro-inflammatory cytokines and causing tissue damage whereas M2 macrophages protect mice by secreting anti-inflammatory factors that aid in the resolution of inflammation 4,14-17. Depending upon the stimulus, monocytes can differentiate into either M1 or M2 macrophages. Our lab previously demonstrated that the TLR2/1 agonist PAM3CSK4 (PAM3) preferentially stimulated normal human and murine monocytes to mature into M2-like macrophages 10,18,19. The murine M2 macrophages co-expressed CD206 and F480. Purified cells of this phenotype were functionally M2 (based on their production of IL-10, phagocytic activity, and inability to produce inflammatory cytokines) 10. By comparison, F480 + macrophages that lacked CD206 were functionally M1 (producing IL-12 and IFNg, lysing tumor targets but lacking phagocytic activity) 18...

show abstract

In Vivo Editing of Macrophages through Systemic Delivery of CRISPR‐Cas9‐Ribonucleoprotein‐Nanoparticle Nanoassemblies

Lee

Mout

Luther

et al. 2019

Advanced Therapeutics

View full text Add to dashboard Cite

Macrophages are key effectors of host defense and metabolism, making them pro mising targets for transient genetic therapy. Gene edit ing through delivery of the Cas9-ribonucleoprotein (RNP) provides mu ltip le advantages over gene delivery-based strategies for introducing CRISP R machinery to the cell. There are, however, significant physiological, cellular, and intracellular barriers to the effective delivery of the Cas9 protein and guide RNA (sgRNA) that have to date, restricted in vivo Cas9 proteinbased approaches to local/topical delivery applications. Herein we describe a new nanoassembled platform featuring co-engineered nanoparticles and Cas9 protein that has been developed to provide efficient Cas9-sgRNA delivery and conco mitant CRISPR edit ing through systemic tail-vein inject ion into mice, achieving >8% gene editing efficiency in macrophages of the liver and spleen.Received: ((will be filled in by the editorial staff))Revised: ((will be filled in by the editorial staff))

show abstract

Regulation of the maturation of human monocytes into immunosuppressive macrophages

Abstract: • NF-kB and Akt regulate human monocyte into macrophage differentiation; p38 MAPK and PTGS2 promote the generation of suppressive macrophage.• The Toll-like receptor 2/1 agonist PAM3 induces human monocytes to mature into immunosuppressive macrophages in vitro and in vivo.

Cited by 24 publications

References 56 publications

PAM3 supports the generation of M2-like macrophages from lupus patient monocytes and improves disease outcome in murine lupus

PAM3 supports the generation of M2-like macrophages from lupus patient monocytes and improves disease outcome in murine lupus

PAM3 protects against DSS-induced colitis by altering the M2:M1 ratio

In Vivo Editing of Macrophages through Systemic Delivery of CRISPR‐Cas9‐Ribonucleoprotein‐Nanoparticle Nanoassemblies

Contact Info

Product

Resources

About