year 2, Issue 3 And 4 (3-2009)
Iran J Med Microbiol 2009, 2(3 And 4): 25-30 |
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Karami S, Siadat S D, Tabaraie B, Norouzian D, Harzandi N, Aghasadeghi M, et al . Evaluation of antibodyresponse to outer membrane proteins of Brucella abortus S99 by Enzyme Linked Immunosorbent Assay. Iran J Med Microbiol 2009; 2 (3 and 4) :25-30
URL: http://ijmm.ir/article-1-125-en.html
URL: http://ijmm.ir/article-1-125-en.html
Sahar Karami1 
, Seyed Davar Siadat2 , Bahman Tabaraie3 , Dariush Norouzian4 , Naser Harzandi1 , Mohammadreza Aghasadeghi3 , Jalal Izadi Mobarakeh3 , Mohammadreza Razavi3 , Mayam Kheirandish5 , Seyed Mehdi Sadat1
, Seyed Davar Siadat2 , Bahman Tabaraie3 , Dariush Norouzian4 , Naser Harzandi1 , Mohammadreza Aghasadeghi3 , Jalal Izadi Mobarakeh3 , Mohammadreza Razavi3 , Mayam Kheirandish5 , Seyed Mehdi Sadat1
1- Department of Microbiology, Isalmic Azad University, Karaj Branch, Iran
2- Bacterial vaccine and antigen preparation unit, Research and Production Complex, Pasteur Institute of Iran, Tehran, Iran ,d.siadat@gmail.com
3- Department of hepatitis and aids, Pasteur Institute of Iran, Tehran, Iran
4- Bacterial vaccine and antigen preparation unit, Research and Production Complex, Pasteur Institute of Iran, Tehran, Iran
5- Research center, Iranian Blood Transfusion Organization
2- Bacterial vaccine and antigen preparation unit, Research and Production Complex, Pasteur Institute of Iran, Tehran, Iran ,
3- Department of hepatitis and aids, Pasteur Institute of Iran, Tehran, Iran
4- Bacterial vaccine and antigen preparation unit, Research and Production Complex, Pasteur Institute of Iran, Tehran, Iran
5- Research center, Iranian Blood Transfusion Organization
Abstract: (16160 Views)
Background and objectives: Currently, antigenic determinants of Brucella cell wall such as outer membrane
proteins (OMPs) and lipopolysaccharide (LPS) are considered as potential candidates to develop subunit
vaccines. It has been shown that OMPs of Brucella abortus along with other bacterial components could
stimulate synthesis of specific IgG antibody in rabbits. The aim of present study was evaluation of antibody
response to OMPs of Brucella abortus S99by ELISA method.
Materials and Methods: OMPs were extracted by deoxycholate extraction technique and further purification performed by sequential centrifugation and ultracentrifugation. Protein concentration determined by using the Nano drop ND – 10000. Sodium dodecyle sulfate polyacryl amid gel electrophoresis (SDS – PAGE) was used to produce a profile of OMPs proteins which these were further differentiated on the basis of molecular weight. By using this method, the class II proteins belongs to porins, which comprised the major OMPs in B. abortus were purified. The groups consisted of 3 rabbits were immunized with three different compounds comprises of OMPs only, combined with LPS and combined with complete Freund's adjuvant (CFA). Sera from immunized animals were then evaluated for specific antibody response.
Results: the most immunogenic compounds were LPS+ porins and CFA+ porins and could promoted specific antibody response in experimental animals. The level of specific antibodies in rabbits immunized with above mentioned combinations was significantly higher compared to immunized rabbits with porins only (P<0.05) .
conclusion: Based on the results, the combination of LPS+ porins can produce long term protective immunity against Brucella. So it could be probably considered for developing of subunit vaccine for prevention of human brucellosis.
Materials and Methods: OMPs were extracted by deoxycholate extraction technique and further purification performed by sequential centrifugation and ultracentrifugation. Protein concentration determined by using the Nano drop ND – 10000. Sodium dodecyle sulfate polyacryl amid gel electrophoresis (SDS – PAGE) was used to produce a profile of OMPs proteins which these were further differentiated on the basis of molecular weight. By using this method, the class II proteins belongs to porins, which comprised the major OMPs in B. abortus were purified. The groups consisted of 3 rabbits were immunized with three different compounds comprises of OMPs only, combined with LPS and combined with complete Freund's adjuvant (CFA). Sera from immunized animals were then evaluated for specific antibody response.
Results: the most immunogenic compounds were LPS+ porins and CFA+ porins and could promoted specific antibody response in experimental animals. The level of specific antibodies in rabbits immunized with above mentioned combinations was significantly higher compared to immunized rabbits with porins only (P<0.05) .
conclusion: Based on the results, the combination of LPS+ porins can produce long term protective immunity against Brucella. So it could be probably considered for developing of subunit vaccine for prevention of human brucellosis.
Type of Study: Original Research Article |
Subject:
Medical Bacteriology
Received: 2013/11/15 | Accepted: 2013/11/16 | ePublished: 2013/11/16
Received: 2013/11/15 | Accepted: 2013/11/16 | ePublished: 2013/11/16
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