Human Immune Response to Polysaccharide-Protein Conjugate Vaccines

Surface polysaccharides of pathogenic bacteria, including capsular polysaccharides (CPS) or lipopolysaccharides (LPS), may serve both as essential virulence factors and as protective antigens. The age-related and T-cell independent immunogenicity of CPS limit their use as vaccines especially in infants and young children. LPS are too toxic to administered. Accordingly, their O-specific polysaccharide (O-SP), that share the virulence promoting and protectiveness of CPS, must be purified: O-SP are too small to be immunogenic (haptens). Covalently binding CPS or of O-SP to medically-useful proteins to form conjugates both increases their immunogenicity and confers T-cell dependence to these saccharides.The O-SP of Shigella sonnei and of Shigella flexneri 2a were bound to bacterial toxoids.

<P>In adults and then in 4-7 year-olds, both conjugates were safe and induced statistically significant and long-lived rises of IgG antibodies to the homologous LPS. Similar, though lesser rises of IgM and IgG anti-LPS were also induced. The levels of serum IgG anti-LPS were equal to or greater that those in recruits that recovered from shigellosis. Re-injection of S. flexneri 2a conjugate induced a booster response in the recruits.

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A Phase 3 trial showed that one injection of S. sonnei O-SP, bound to a non-toxic recombinant Pseudomonas aeruginosa exoprotein A (rEPA) protected army recruits against outbreaks with this pathogen. Importantly, there was a statistically-significant correlation between the levels of serum IgG anti-LPS and the efficacy of the conjugate.

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Two methods were developed that increased the immunogenicity of the Shigella conjugates in mice. First another carrier protein, a genetically-inactivated Corynebacterium diphtheriae toxin (CRM9) was a superior carrier for S. sonnei O-SP. Second treatment of rEPA with succinic anhydride, a non-toxic mild akylating agent that converts amino groups of proteins to carboxyls, increased the immunogenicity of S. flexneri 2a O-SP.
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A phase 1 study of these improved Shigella conjugates confirmed their immunogenicity, and these new products are being evaluated for their clinical efficacy in a Phase 3 trial in Israel. In collaboration with the Lanzhou Vaccine Institute and Provincial Medical Center in Henan,China, a clinical trial of these two conjugates is being planned. A double mutant of Bordetella pertussis, producing a genetically-inactivated toxin and deficient in FHA synthesis was developed. Effort is directed towards increasing production of this B. pertussis strain as a more easily purified pertussis toxin for a monocomponent vaccine and as a carrier protein for pneumococcal type 14 CPS. Clostridium difficile is a major cause of hospital-acquired diarrhea following antibiotic usage: the diarrhea is mediated by two exotoxins, A and B. Toxin A, considered to be the major toxin, in its extreme form it will cause pseudomembranous colitis. A genetically-derived toxin mutant (rARU) induces both antitoxin and protects animals from infection with C. difficile. The solubility of rARU improved its solubility and did not detectable affects its reaction with antiserum. Techniques to prepare the mutant toxins A and B for clinical use have been worked out. Three polysaccharide of varying composition, pneumococcus type 14, Escherichia coli K1 and S. flexneri 2a were conjugated to succinylated rARU. The resultant conjugates induced high levels of both anti-polysaccharide and antitoxin. Clinical evaluation of these conjugates, designed to protect two diseases, is underway.