Abstract

THE EFFICIENCY OF EXTRACELLULAR POLYMERIC SUBSTANCE EXTRACTED FROM PANTOEA AGGLOMERANS IN ACTIVATION IMMUNE RESPONSE

Layla S. Abdulhassan and *Mayyada F. Darweesh

Abstract

During the period from December 2014 to July 2015, 24isolates of Pantoea agglomerans were founded to be associated with infection like hemodialysis inflammation(3 isolates), chronic diarrhea (3isolates), UTI(2isolates), otitis media (3), burn (5), wound (2), Also 6 isolates from environmental hospital from AL-Hakeem Hospital and Al-Sadar Medical City in AL-Najaf province, All isolates were identify by morphology properties after culturing on MacConkey and Blood agar, conventional biochemical test and carbohydrate fermentation then finally by using Vitek GN-card. All isolates were examined to choose the most efficient ones in the formation of biofilm and select it to extract EPS. The result illustrated that P.agglomerans No.4 had greater biofilm formation (0.816) and elected to EPS extraction which divided into two type Bound-EPS that gave protein concentration 486.12 g/ml, Carbohydrate concentration 72.21 g/ml while second type Free-EPS with protein concentration 247.6 g/ml, Carbohydrate concentration 56.42 g/ml. Bound-EPS and Free-EPS used as immunogenes to stimulate immune system by injected the Rats intraperitonealy with 1ml of LD50 % from both immunogenes then the efficiency of immunogenes in stimulation the immune response has been evaluated by determine the total and differential count of WBC and Cytokines(IL-4 and IL-12) levels in addition to determined Arthus and DTH. The total count of WBC has shown the significant increase compared to control group in the rats treated with Bound-EPS and Free-EPS, The third dose gave maximum WBC count 13431.26 and11834.33cells/ mm3 respectively, due to the elevated count of neutrophils and lymphocytes. The results of Arthus reaction revealed 1.43 mm of Bound-EPS and 1.321mm of Free-EPS compared with control 0.54 mm, also the higher mean of DTH reaction was revealed of Bound-EPS and Free-EPS compared to control group. The level of IL-4 estimated by ELISA and show that there was a significant increase compared with the control group in the rats treated with second and third dose of Bound-EPS and third dose of Free-EPS (2.510, 2.532, 1.984)pg/ml respectively, The results also reveal that the concentration of IL-12 is significantly higher in the serum of rats treated with Bound-EPS and Free-EPS as compared with control group. In conclusion Bound –EPS more efficient in stimulating immune response than Free-EPS and can gave protection against most biofilm infection. P.agglomerans, Extracellular polymeric substances, vaccine, immune system.

Keywords: P.agglomerans, Extracellular polymeric substances, vaccine, immune system.