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CONSTRUCTION OF LIVE ATTENUATED SHIGELLA VACCINE STRAINS THAT EXPRESS CFA/I ANTIGENS (CFAB AND CFAE) AND THE B SUBUNIT OF HEAT-LABILE ENTEROTOXIN (LTB) FROM ENTEROTOXIGENIC E. COLI

With the goal of creating a combination vaccine against Shigella and other diarrheal pathogens we have constructed a prototype vaccine strain of Shigella flexneri 2a (SC608) that can serve as a vector for the expression and delivery of heterologous antigens to the mucosal immune system. SC608 is an asd derivative of SC602, a well-characterized vaccine strain, which has recently undergone several phase 1 and 2 trials for safety and immunogenicity. Using non-antibiotic asd-based plasmids, we have created novel constructs for the expression of antigens from enterotoxigenic E. coli (ETEC), including CFA/I (CfaB and CfaE) and the B-subunit from heat-labile enterotoxin (LTB) in Shigella vaccine strain SC608. Heterologous protein expression levels and cellular localization are critical to immune recognition and have been verified by immunoblot analysis. Following intranasal immunization (SC608(CFAI) and SC608(CFAI/LTB) of guinea pigs, serum IgG and IgA immune responses to both the Shigella LPS and ETEC antigens can be detected by ELISA. In addition, ELISPOT analysis for ASCs from cervical lymph nodes and spleen showed similar responses. All vaccine strains conferred high levels of protection against challenge with wild-type S. flexneri 2a using the Sereny test. Furthermore, serum from guinea pigs immunized with SC608 expressing CfaB and LTB contained antibodies capable of neutralizing the cytological affects of heat-labile toxin (HLT) on Chinese Hamster Ovary (CHO) cells. These initial experiments demonstrate the validity of a multivalent invasive Shigella strain that can serve as a vector for the delivery of pathogen-derived antigens.


Docket:WRAIR 2004-35

Publication/Issued No.:7,759,106

Publication/Issue Date:2010-07-20

Categories: Vaccine

More Detail:Visit USPTO.GOVExternal link

Lab:WRAIR

Inventor(s):RANALLO, R.; VENKATESAN, M.


For Licensing Opportunities, please contact the ORTA at USArmy.Detrick.MEDCOM-USAMRMC.List.ORTA@mail.mil
 

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