Background/Purpose: Genetic mutations in key regulatory molecules of the innate immune system cause autoinflammatory diseases through propagation of hyperinflammatory responses. Monocytes/ macrophages regulate inflammatory processes during infection and homeostasis by secreting a wide range of cytokines and chemokines that control recruitment and migration of immune cells to inflamed tissue, and their biological functions through immune cell differentiation and polarization. We characterized the cytokine/chemokine profile of peripheral blood monocyte-derived M1-, and M2-macrophages (M1-, M2-MDMs) from patients (pts) with genetically defined systemic autoinflammatory diseases (SAIDs) and controls.

Methods: Whole blood from pts enrolled in the IRB approved study (NCT 02974595) with IL-1 mediated  SAIDs NOMID (n=3), DIRA (n=4), Majeed Syndrome (n=1); interferon-mediated diseases CANDLE (n=3) SAVI (n=3); IL-18 mediated diseases NLRC4-MAS (n=2), IL-18PAP-MAS(n=1), Blau syndrome (n=4), NEMO-NDAS (n=3); and healthy controls (HC) (n=6) were assessed. M1-, M2-MDMs) from culture with GM-CSF or M-CSF for 6 days were assessed with and without LPS+ATP stimulation. Human Cytokine Array and ELISA were used for cytokine/chemokine profiling. SAVI monocytes died upon in vitro M1-M2 MDM differentiation. Differentiation into rounded and flat shaped M1-MDMs and elongated or spindle shaped M2-MDMs was 80-90% and 70-80% respectively for all samples.

Results: IL-1β secretion from M1-MDMs was higher in NOMID, DIRA and Majeed syndrome pts upon stimulation with LPS and ATP compared to HC and other SAID pts. In NOMID pts, IL-1b was equally elevated in M1-and M2 MDMs, whereas IL-18 was produced in M1- and M2-MDMs in NLRC4-MAS and in M2-MDMs in IL-18PAP-MAS. TNF and IL-6 were highest in stimulated M1-MDMs in NOD2 and NLRC4 MAS pts. compared to HC (p< 0.05) and in stimulated M2-MDMs in Majeed, CANDLE, NOD2 and NLRC4 pts. Among chemokines, CCL2/MCP-1 which recruit monocyte into inflamed tissues was constitutively expressed in M1/M2-MDMs of CANDLE and NOD2 and M1-MDM of NLRC4-MAS pts. M2-MDMs derived from Majeed and DIRA pts. produced significantly higher levels of CCL5/RANTES and CXCL10/IP-10 (p< 0.05). IP-10 secretion was not seen in CANDLE or NEMO-NDAS. IL-8 and CXCL1, two neutrophil-attracting chemokines were high in most autoinflammatory disease groups, moreover, M2-MDMs of CANDLE and Majeed pts constitutively released IL-8. As expected, IL-1ra was absent in unstimulated and stimulated DIRA M1-, and M2-MDSs. Moreover, CANLDE MDMs did not show an upregulation of IL-1ra distinct from IL-1 mediated diseases.

Conclusion: Taken together, differentiation of monocytes into M1 and M2 MDMs leads to variable but disease-specific regulation of pro-inflammatory cytokine and chemokine production. The value of this system in assessing monocyte and macrophage polarization defects and in assessing the proinflammatory dysregulation in various SAIDs need to be further explored.  Particularly, whether such systems proof useful in conditions where more than one cytokine/chemokine are dysregulated and in suggesting treatment targets needs to be evaluated in the future.

This work was supported by the DIR, NIAID, NIH.

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ACR Meeting Abstracts - https://acrabstracts.org/abstract/characterization-of-cytokine-chemokine-profile-in-patient-derived-m1-m2-macrophages-to-identify-biomarkers-for-genetically-defined-systemic-autoinflammatory-diseases/