Masooma Abdullahi; Reza Ranjbaran; Soheaila Alyasin; Zeinab Keshavarz; Amin Ramezani; Abbas Behzad-Behbahani; Sedigheh Sharifzadeh
Volume 13, Issue 1 , March 2016, , Pages 27-36
Abstract
Background: Asthma is very common in children and its diagnosis is based on clinical manifestations, which can be misdiagnosed as other respiratory diseases with similar signs and symptoms. Objective: To analyze the expression of ST2L and CD203c in the diagnosis of pediatric asthma. Methods: Basophils ...
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Background: Asthma is very common in children and its diagnosis is based on clinical manifestations, which can be misdiagnosed as other respiratory diseases with similar signs and symptoms. Objective: To analyze the expression of ST2L and CD203c in the diagnosis of pediatric asthma. Methods: Basophils were purified from whole blood samples of patients and healthy controls using Ficol-Paque gradient and Basophil Isolation Kit. RNA extraction was done by RNX-Plus solution and after synthesis of cDNA, the gene expression was analyzed by means of real time PCR. Results: Patients expressed significantly higher levels of CD203c than healthy controls (p=0.01). Although there was an increase in the transcription level of ST2L gene in patients, the results were not statistically significant compared to those obtained from the healthy controls (p>0.05). A Specificity of 60% and a sensitivity of 73% were foundusing ROC curve for CD203c expression. Patients with positive family history of asthma exhibited more CD203c and ST2L expression (p<0.05). Conclusion: It is proposed that determining CD203c expression by real time PCR may be an effective technique for diagnosis of pediatric asthma.
Esmaeil Sadroddiny; Jafar Ai; Kathleen Carroll; Trong Khoa Pham; Phillip Wright; Ashutosh Pathak; Birgit Helm
Volume 9, Issue 1 , March 2012, , Pages 1-31
Abstract
Background: Secretory proteins of IgE receptor activated mast cells and basophils play a pivotal role in the generation of immediate and long term immune responses in allergy and type I hypersensitivity. Objective: The present study aims to generate a 2-D map and profile of proteins secreted from a high ...
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Background: Secretory proteins of IgE receptor activated mast cells and basophils play a pivotal role in the generation of immediate and long term immune responses in allergy and type I hypersensitivity. Objective: The present study aims to generate a 2-D map and profile of proteins secreted from a high secretory variant of the rat basophilic leukemia cell line, RBL-2H3.1, which in view of the difficulty associated with gaining adequate numbers of pure primary mast cell and basophiles, represents an accepted model system for the study and standardization of the methodology to characterize the secretome of these cell types. Methods: A 2-D map of secretory proteins was generated by 2-D PAGE and a shotgun mass spectrometric approach carried out for protein identi fication. Results: Study resulted into identification of 299 proteins released from resting and IgE receptor activated RBL-2H3.1 cells after 90 s, 30 min and 3 h antigen challenge. Further sequence analysis identified ~53% of total proteins as secretory proteins which could be attributed to classical and non-classical secretory pathways. Additionally, functional classification of classic secretory proteins verified the presence of proteins belonged to cytokines, receptors, membrane proteins, lysosomal proteins and proteins associated with specific sub-cellular localizations such as endoplasmic reticulum, mitochondria, nucleus, cytoplasm and ribosome. According to this data the presence of some secretory proteins such as cytokines (e.g. MCP-2, PF-4, CSF-1 and TGF-β1) are all subject to Ag challenge which may point to their importance toward pathogenesis in allergic diseases. Conclusion: In view of both a beneficial and adverse role of mast cell mediators in health and disease, an identification of temporal changes in the secretory pattern may form the basis for future tailor made intervention strategies that may enable us to harvest the therapeutic potential inherent in mast cell exocytosis while inhibiting/attenuating negative outcomes.
