Phagocytosis via complement receptor 3 enables microbes to evade killing by neutrophils.
Asya SmirnovKylene P DailyMary C GrayStephanie A RaglandLacie M WernerM Brittany JohnsonJoshua C EbyErik L HewlettRonald P TaylorAlison K CrissPublished in: Journal of leukocyte biology (2023)
Complement receptor 3 (CR3; CD11b/CD18; αmβ2 integrin) is a conserved phagocytic receptor. The active conformation of CR3 binds the iC3b fragment of complement C3 as well as many host and microbial ligands, leading to actin-dependent phagocytosis. There are conflicting reports about how CR3 engagement affects the fate of phagocytosed substrates. Using imaging flow cytometry, we confirmed that binding and internalization of iC3b-opsonized polystyrene beads by primary human neutrophils was CR3-dependent. iC3b-opsonized beads did not stimulate neutrophil reactive oxygen species (ROS), and most beads were found in primary granule-negative phagosomes. Similarly, Neisseria gonorrhoeae (Ngo) that does not express phase-variable Opa proteins suppresses neutrophil ROS and delays phagolysosome formation. Here, binding and internalization of Opa-deleted (Δopa) Ngo by adherent human neutrophils was inhibited using blocking antibodies against CR3 and by adding neutrophil inhibitory factor, which targets the CD11b I-domain. No detectable C3 was deposited on Ngo in the presence of neutrophils alone. Conversely, overexpressing CD11b in HL-60 promyelocytes enhanced Δopa Ngo phagocytosis, which required CD11b I domain. Phagocytosis of Ngo was also inhibited in mouse neutrophils that were CD11b-deficient or treated with anti-CD11b. Phorbol ester treatment upregulated surface CR3 on neutrophils in suspension, enabling CR3-dependent phagocytosis of Δopa Ngo. Neutrophils exposed to Δopa Ngo had limited phosphorylation of Erk1/2, p38, and JNK. Neutrophil phagocytosis of unopsonized Mycobacterium smegmatis, which also resides in immature phagosomes, was CR3-dependent and did not elicit ROS. We suggest that CR3-mediated phagocytosis is a silent mode of entry into neutrophils, which is appropriated by diverse pathogens to subvert phagocytic killing.
Keyphrases
- reactive oxygen species
- cell death
- endothelial cells
- signaling pathway
- nk cells
- flow cytometry
- dna damage
- binding protein
- mycobacterium tuberculosis
- transcription factor
- high resolution
- oxidative stress
- microbial community
- induced pluripotent stem cells
- multidrug resistant
- molecular dynamics simulations
- adverse drug
- dna binding
- crystal structure