optimized method for ex vivo generation of dendritic cells

Dendritic cells are the most potent antigen presenting cells. They capture, process and present antigens to T cells and secrete various cytokines and soluble factors to initiate adaptive immune responses. Dendritic cells are also important in induction of immunological tolerance to self- antigens. Due to their crucial role in immune responses during infections, cancers, transplantations, allergies, and autoimmune diseases, they have become important targets for many biological and clinical studies. Usually large numbers of dendritic cells is essential for these studies; however, small numbers of these cells in blood and tissues makes the isolation very difficult. Therefore, In vitro generation of these cells is useful for research and clinical applications. The aim of this study was in vitro generation of dendritic cells from bone marrow-hematopoietic progenitor cells using a simple and efficient method. Murine bone marrow cells were cultured in medium supplemented with Granulocyte-macrophage colony-stimulating factor and Interleukin 4 without depletion of any cell population for 9 days. Fresh medium containing cytokines was added every 3 days. Analysis of the morphology and immunophenotype of cultured cells showed the generation of dendritic cells from day 2 of the culture period. But, the number of cells that possess morphological characteristics and typical cell surface markers [CD11c, MHC-II and CD86] of dendritic cells was elevated by increasing the culture period. The purity of dendritic cells was 86% by the end of 9 days culture. This method can be used as an efficient method for ex vivo generation of dendritic cells

[Comparative evaluation of PCR and standard serological methods in detection of brucellosis in cattle]

Brucellosis is one of the most dangerous infectious zoonotic diseases which at present affect most areas of Iran directly or indirectly. The aim of the present study was evaluate the efficacy of PCR test in detection of brucellosis. Suspected animal sera [n=20] to brucellosis were collected from clinics and subjected to Rose Bengal and Wright Agglutination tests and to PCR analysis. While 6 samples [30%] were positive with titer 1:80. To confirm, all samples were subjected to PCR which revealed 5 samples of 1:80 and 6 samples of less than 1:80 were in agreement with wright test. However, 8 samples of 1:80 and 1 sample of less than 1:80 were in disagreement with wright test. The results of this study revealed that considerable percent of submitted samples were positive for brucellosis and positivity of ones were confirmed with PCR test we have shown 55% agreement between SAT [Standard Agglutrination Test] and PCR results

Detection of anti-E coli, rota virus and corona virus antibodies in sera samples of diarrheic and normal calves under 1 month of age

The aim of the present study was to evaluate the presence anti- E. coli, -rotavirus and -corona virus, in calves' sera. A total of 184 calves under 1 month of age [84 diarrheic and 100 normal] was studied. Serological tests including: direct ELISA for detection of anti-K99 E. coli, - rotavirus and - corona virus and tube agglutination test for detection anti-O157 E. coli, antibodies were used. Data were analyzed by chi-square, fisher test and t-student tests. Anit-K99 E. coli antibodies were detected in 56% and 66% of diarrheic and normal calves, respectively. Tube agglutination test showed the presence of anti- O157-E. coli antibodies in 82% and 69% of diarrheic and normal calves respectively. Anti-rotavirus antibodies were detected in 100% and 99% of diarrheic and normal calves, respectively. Anti- corona virus antibodies were detected in 82% and 72% of diarrheic and normal calves respectively. Conclusion: The results of the present study may indicate the high exposure of the examined dairy cattle population to E. coli, rotavirus and corona virus and also the absence of correlation between such serological responses with the prevention of calves' diarrhea