SOCS3 Silencing Attenuates Eosinophil Functions in Asthma Patients
Abstract
:1. Introduction
2. Results
2.1. SOCS3 Down-Regulation in Blood Eosinophils from Asthmatic Patients by siRNA
2.2. Immune Response Elements Altered by Interfering SOCS3 Expression in Eosinophils from Asthmatic Subjects
2.3. Eosinophil Migration Suppressed by SOCS3 Gene Silencing
2.4. SOCS3 Gene Silencing Reduced Eosinophil Adhesion
2.5. Eosinophil Degranulation Recovery after SOCS3 Silencing
2.6. JAK-STAT but not MAPK/ERK Pathway Is Affected by SOCS3 Down-Regulation in Eosinophils from Asthmatics
3. Discussion
4. Experimental Section
4.1. Subjects
Items | Subjects |
---|---|
n | 36 |
Age (years) * | 46 (20–86) |
Male (%) | 11 (28.2) |
Atopy (%) | 29 (74.2) |
FEV1 (%) * | 100 (73–138) |
FEV1/FVC (%) * | 79 (62–100.4) |
FeNO ppb * | 31 (8.5–230) |
Peripheral eosinophilia (%) * | 4.7 (1.5–19.1) |
4.2. Recombinant Proteins and Reagents
4.3. Eosinophil Isolation
4.4. Eosinophil Culture
4.5. RNA Isolation, RT-PCR, and TaqMan Gene Expression Assays
4.6. Fluorescence Microscopy
4.7. Eosinophil Migration
4.8. Eosinophil Adhesion
4.9. Eosinophil Degranulation
4.10. Flow Cytometry
4.11. Immunoblot
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Zafra, M.P.; Cañas, J.A.; Mazzeo, C.; Gámez, C.; Sanz, V.; Fernández-Nieto, M.; Quirce, S.; Barranco, P.; Ruiz-Hornillos, J.; Sastre, J.; et al. SOCS3 Silencing Attenuates Eosinophil Functions in Asthma Patients. Int. J. Mol. Sci. 2015, 16, 5434-5451. https://doi.org/10.3390/ijms16035434
Zafra MP, Cañas JA, Mazzeo C, Gámez C, Sanz V, Fernández-Nieto M, Quirce S, Barranco P, Ruiz-Hornillos J, Sastre J, et al. SOCS3 Silencing Attenuates Eosinophil Functions in Asthma Patients. International Journal of Molecular Sciences. 2015; 16(3):5434-5451. https://doi.org/10.3390/ijms16035434
Chicago/Turabian StyleZafra, Mª Paz, Jose A. Cañas, Carla Mazzeo, Cristina Gámez, Veronica Sanz, Mar Fernández-Nieto, Santiago Quirce, Pilar Barranco, Javier Ruiz-Hornillos, Joaquín Sastre, and et al. 2015. "SOCS3 Silencing Attenuates Eosinophil Functions in Asthma Patients" International Journal of Molecular Sciences 16, no. 3: 5434-5451. https://doi.org/10.3390/ijms16035434