Paper Information

Journal:   TEHRAN UNIVERSITY MEDICAL JOURNAL (TUMJ)   2004 , Volume 62 , Number 5; Page(s) 363 To 371.
 
Paper: 

DETECTION OF INTRACELLULAR CYTOKINE EXPRESSION IN WHOLE BLOOD PREPARATIONS FROM NORMAL AND PATIENTS WITH BRUCELLOSIS

 
 
Author(s):  RAFIEE A.R., KEIHANI A.H., KARIMINIA AMINA, KABOUDANIAN ARDESTANI S., MOHRAZ M., AMIRKHANI A.
 
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Abstract: 

Background: Human brucellosis is a orldwide zoonotic infection caused by intracellular bacteria belong to the genus of Brucella. Based on the murine studies it has been shown that host resistance to Brucella depends on Th1 response, whereas Th2 response is involved in the severity of disease. The analysis of cytokine production is a valuable component of studies of immune response to stimulation such as pathogens, vaccines, and other immunological challenges. This article is introduced a simple, rapid throughput of clinical samples and a cheap and reliable assay.
Materials and Methods: For this technique to be useful in clinical setting, we have used un-separated whole blood of healthy normal and patients with acute and chronic brucellosis as the source of cells evaluation of interferon gamma (IFN-
g) and interleukin-13 expression. Diluted whole blood samples of patients with brucellosis (n=15) and, sex and age-matched healthy volunteers (n=14) were cultured in the presence of either mitogen; heat inactivated bacteria or medium alone. IL-13 and IFN- g were measured by specific sandwich ELISA and flow cytometry was detected the number of cytokine-producing CD3+ cells.
Results: It was found that not only IFN-
g production but also the number of IFN-g producing CD3 cells were significantly decreased in response to antigen in chronic group of patients. There was a reverse correlation between the number of IFN-g producing and CD3 IL-13-producing cells only in acute group which shown polarization of immune responses to Thl in them. Although the percentage of CD3 IL-13-producing cells was dramatically high in the chronic group of patients no correlation was found between the number of CD3 IFN-€-producing and CD3 IL-13- producing cells.
Conclusion: In conclusion, the correlation of Th2 cytokines production and progression of chronic human brucellosis was not demonstrated. Nevertheless, diminished production of Th1 cytokines production in chronic group may suggest T cells unresponsiveness to Brucella antigen which helps prolongation of brucellosis in chronic patients.

 
Keyword(s): HUMAN BRUCELLOSIS, WHOLE BLOOD, IFN-Y, IL-13
 
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