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Isolation, Transfection, and Culture of Primary Human Monocytes

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Monocyte Dll4 Contributes to Systemic Inflammation in People Living With HIV.

Shumin Wang¹, Meera V Singh², Craig N Morrell¹

¹Aab Cardiovascular Research Institute and Department of Medicine, University of Rochester School of Medicine and Dentistry, NY (S.W., C.N.M., J.R., T.W., L.A.M., J.J.X., T.N., S.T., J.P.).

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|June 12, 2026

Summary

Monocyte delta-like 4 (Dll4)-Notch signaling drives inflammation in people living with HIV (PLWH) by suppressing antiviral responses and increasing monocyte migration. Monocyte-derived Dll4 is a potential therapeutic target for systemic inflammation in PLWH.

Keywords:

endothelial cells gene expression inflammation lipopolysaccharides monocyte

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Area of Science:

Immunology
Cell Biology
Molecular Biology

Background:

Persistent systemic inflammation in people living with HIV (PLWH) contributes to comorbidities like cardiovascular disease.
Monocyte-mediated inflammation is a key driver of chronic inflammation in PLWH, but the mechanisms are not fully understood.
This study investigates the role of delta-like 4 (Dll4)-Notch signaling in monocyte-driven inflammation.

Purpose of the Study:

To investigate the role of Dll4-Notch signaling in monocyte-mediated inflammation in PLWH.
To determine the mechanisms by which Dll4 influences inflammatory responses and monocyte behavior.
To identify Dll4 as a potential biomarker and therapeutic target for systemic inflammation.

Main Methods:

Quantified monocyte membrane-bound Dll4 (mDll4) and plasma extracellular Dll4 (exDll4) in PLWH.
Analyzed Dll4-Notch1 signaling activation and its effects on inflammatory gene expression, monocyte adhesion, and transmigration in vitro.
Utilized RNA sequencing after Dll4 silencing to identify downstream signaling pathways.

Main Results:

PLWH showed increased mDll4 expression and Dll4-Notch1 signaling activation.
Plasma exDll4 was elevated in males with HIV, correlating with monocyte mDll4 and proinflammatory monocytes.
Dll4-Notch signaling suppressed interferon-related antiviral responses and enhanced monocyte adhesion and transmigration.

Conclusions:

Proinflammatory signals upregulate monocyte Dll4, activating Dll4-Notch signaling.
This signaling suppresses antiviral responses and promotes monocyte transmigration, sustaining inflammation in PLWH.
Monocyte-derived Dll4 represents a promising biomarker and therapeutic target for managing systemic inflammation.