Abstract
Cholera is toxin-mediated enteroinfection, with epidemic character and there are approximately 120000 death cases per year worldwide. Protection against cholera has not been accomplished due to deficiencies in the licensed vaccines. Serum vibriocidal activity mediated by LPS antibodies is the only immune segment correlated with the resistance of cholera. On the basis of literature data (Robbins JB, 1990; Ogawa Y, 1996) we synthesized glucoconjugates, composed of detoxified LPS from Vibrio cholerae and protein carriers. Conjugate vaccines were prepared by binding acetic acid and hydrazine-treated lipopolysaccharide (LPS) from Vibrio cholerae O1, serotype Inaba, to cholera toxin B-subunit (CT-B) and bovine serum albumin (cBSA). Adipic acid dihydrazide was used for derivatization of oligosaccharides and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) as conjugating agent. SDS-PAGE, glycoprotein detection and TLC dot-blot were used for physical and chemical analysis of the prepared four types of conjugates. Safe level of endotoxins, measured by LAL assay was detected in all conjugates. The synthesized conjugates can be used for monitoring immunization schemes on experimental animals. It is to be expected that conjugated vaccines are safe and efficient and that will have high immunogenic and T-dependant characteristics with long immune protection against cholera.
Highlights
Cholera is caused by Vibrio cholerae strains belonging to two serogroups, O1 and O139, based on their LPS structure
In the year 2002, World Health Organization (WHO) estimated that over 220,000 people were stricken with cholera each year
LPS as main bacterial antigen determinant with immunogenic characteristics will be used for synthesis of glycoconjugates
Summary
Cholera is caused by Vibrio cholerae strains belonging to two serogroups, O1 and O139, based on their LPS structure. Serogroup O1, which is responsible for the seventh cholera pandemic, is divided into three serotypes, Inaba, Ogawa and Hikojima [1]. In the year 2002, World Health Organization (WHO) estimated that over 220,000 people were stricken with cholera each year. Natural infection and currently available vaccines offer incomplete protection of relatively short duration. The long-term control of cholera depends on good personal hygiene, uncontaminated water supply and appropriate sewage disposal. The improvement of hygiene is a distant goal for many countries. The availability of an effective cholera vaccine
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