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Eosinophil major basic protein enhances the expression of neutrophil CR3 and p150,95
J N Moy1, L L Thomas, L C Whisler
1Department of Immunology/Microbiology, Rush Medical College, Chicago, IL 60612-3864.
Background:
We examined the effect of major basic protein (MBP) stimulation on the expression of the neutrophil B2-integrins: LFA-1 (CD11a/CD18), CR3 (CD11b/CD18), and p150,95 (CD11c/CD18).
Methods:
Incubation of neutrophils with 0.75 to 3.0 mumol/L MBP for 30 minutes at 37 degrees C resulted in concentration-dependent increases in CR3 expression as detected by staining with the CD11b-specific monoclonal antibody, Leu-15.
Results:
The expression of CR3 was significantly (p < 0.001) higher when neutrophils were stimulated with 1.5 mumol/L (53% +/- 9% increase) or 3.0 mumol/L (100% +/- 17% increase) MBP as compared with unstimulated neutrophils. The kinetics of MBP-stimulated CR3 expression were rapid and were similar to those of 100 nmol/L. N-formyl-methionyl-leucyl-phenylalanine-stimulated enhancement of CR3 expression. Incubation of neutrophils with reduced and alkylated MBP resulted in significantly lower (p < 0.05) increases in CR3 expression as compared with stimulation with native MBP. In addition, neither eosinophil cationic protein nor eosinophil-derived neurotoxin altered neutrophil CR3 expression. MBP stimulated minimal increases in LFA-1 expression. However, staining with the monoclonal antibody Leu-M5 (anti-CD11c) revealed that MBP stimulated significant increases in the expression of p150,95.
Conclusions:
These results indicate that MBP stimulates the increased expression of neutrophil adhesion molecules, which in turn may enhance the inflammatory role of neutrophils in the late-phase events of allergic diseases.
Insights
Major basic protein (MBP) significantly increases neutrophil CR3 and p150,95 expression, enhancing their inflammatory role in allergic diseases. This study reveals MBP
Area of Science:
- Immunology
- Cell Biology
- Allergy Research
Background:
- Neutrophils play a key role in inflammatory responses.
- Neutrophil B2-integrins, including LFA-1, CR3, and p150,95, are crucial for immune cell adhesion and function.
- Major basic protein (MBP) is a key mediator released by eosinophils during allergic inflammation.
Purpose of the Study:
- To investigate the effect of major basic protein (MBP) on the expression of neutrophil B2-integrins.
- To determine how MBP influences the adhesion molecules LFA-1, CR3, and p150,95 on neutrophils.
Main Methods:
- Neutrophils were incubated with varying concentrations of MBP.
- Flow cytometry and monoclonal antibody staining (Leu-15 for CD11b, Leu-M5 for CD11c) were used to quantify B2-integrin expression.
- Comparisons were made between native MBP, reduced/alkylated MBP, and other eosinophil granule proteins.
Main Results:
- MBP significantly increased CR3 expression in a concentration-dependent manner.
- MBP-induced CR3 expression showed rapid kinetics, similar to N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation.
- MBP also significantly increased p150,95 expression, with minimal effect on LFA-1.
- Native MBP was more effective than reduced/alkylated MBP in increasing CR3 expression.
Conclusions:
- MBP stimulates the expression of neutrophil adhesion molecules CR3 and p150,95.
- This upregulation of adhesion molecules may contribute to the inflammatory role of neutrophils in late-phase allergic reactions.
- MBP is identified as a key factor in modulating neutrophil adhesion during allergic responses.
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