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Cell-specific, activation-dependent regulation of neutrophil CD32A ligand-binding function
S Nagarajan1, K Venkiteswaran, M Anderson
1Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Georgia Institute of Technology, Atlanta, GA 30322, USA.
Insights
Neutrophils regulate immune responses by switching the affinity of FcgammaRIIA (CD32A) from low to high upon activation. This allows selective engagement of Fc receptors for efficient function.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Neutrophils express two low-affinity Fc gamma receptors (FcγRs): FcγRIIIB (CD16B) and FcγRIIA (CD32A).
- These receptors differ in membrane anchoring and signaling capabilities, but the biological significance of their coexpression is unclear.
- Understanding FcγR regulation is crucial for comprehending neutrophil-mediated immune functions.
Purpose of the Study:
- To investigate the functional significance of coexpressed FcγRIIIB and FcγRIIA on neutrophils.
- To determine if neutrophils modulate FcγR affinity to regulate their functions.
- To elucidate the distinct roles of CD16B and CD32A in neutrophil immune complex binding and signaling.
Main Methods:
- Utilized neutrophils from a CD16B-deficient donor and normal neutrophils treated with anti-CD16 monoclonal antibodies.
- Assessed neutrophil rosetting with sheep erythrocytes opsonized with rabbit IgG (EA).
- Measured FcγR-dependent immunophagocytosis and immune complex (IC) binding.
- Investigated CD32A function on Chinese hamster ovary (CHO) cells for comparison.
Main Results:
- CD16B-deficient neutrophils exhibited poor EA rosetting and impaired immunophagocytosis.
- Neutrophil activation with fMLP significantly increased CD32A-dependent EA rosetting and IC binding in both CD16B-deficient and normal neutrophils.
- This affinity modulation of CD32A was specific to neutrophils and not observed in CHO cells expressing CD32A.
Conclusions:
- Resting neutrophils primarily utilize CD16B for immune complex binding, with CD32A in a low-affinity state.
- Upon activation, neutrophils convert CD32A to a high-affinity state, enhancing CD32A-dependent ligand binding and signaling.
- Neutrophils employ a novel strategy of selective FcγR engagement via affinity modulation for efficient immune functions.
Abstract:
Neutrophils express 2 low-affinity FcgammaR, FcgammaRIIIB (CD16B), and FcgammaRIIA (CD32A). CD16B is a glycosyl-phosphatidyl inositol-anchored molecule, whereas CD32A is a polypeptide-anchored molecule. These 2 receptors also differ in their signaling. The biological significance of coexpression of 2 FcgammaRs with distinct membrane anchors and signaling capacities is not clearly understood. Using neutrophils from a CD16B-deficient donor and normal neutrophils treated with anti-CD16 monoclonal antibodies, the authors demonstrated that affinity modulation of CD32A is one of the mechanisms by which neutrophils regulate their FcgammaR-dependent functions. Neutrophils isolated from a CD16B(- )donor rosetted poorly with sheep erythrocytes opsonized with rabbit IgG (EA) (12% +/- 2% versus 80% +/- 6% for control) and were unable to mediate immunophagocytosis. However, activation of CD16B(-) neutrophils with fMLP, a bacterial chemotactic peptide, increased the CD32A-dependent EA rosetting to 58%. The CD32A-dependent rosetting of fMLP-activated normal neutrophils also increased nearly 5-fold, but there was no increase in CD32A expression. The CD32A-dependent immune complex (IC) binding was also increased in activated neutrophils. This affinity regulation was not observed with CD32A expressed on Chinese hamster ovary cells. These results suggest that in resting neutrophils CD32A is in a low-affinity state and that these cells primarily engage CD16B for IC binding. However, once the neutrophils are activated, the CD32A is converted to a high-affinity state that leads to CD32A-dependent ligand binding and signaling. These results suggest that neutrophils adopt a novel strategy to engage the 2 different FcgammaR selectively during physiologic and pathologic conditions to carry out their functions efficiently.
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