Abstract
Cholera remains to be a global health problem without suitable vaccines for endemic control or outbreak relief. Here we describe a new parenteral vaccine based on neoglyco-conjugate of synthetic fragments of O-specific polysaccharide (O-SP) of Vibrio cholerae O1, serotype Ogawa. Hexa-, octa- and decasaccharides of the O-SP with carboxylic acid at the reducing end were chemically synthesized and conjugated to tetanus toxoid (TT). The conjugates prepared by a novel linking scheme consisted of 17-atom linker of hydrazide and alkyl bonds elicited robust serum IgG anti-LPS responses with vibriocidal activities in mice. There is a length dependence in immune response with decasaccharide conjugates elicited the highest anti-LPS IgG. There seems to be an indication that regardless of the carbohydrate chain length, a molar ratio of 230 ± 10 monosaccharide units per TT induced high antibody response. The conjugates also elicited cross-reactive antibodies to serotype Inaba. The formulation of the proposed cholera conjugate vaccine, similar to other licensed polysaccharide vaccine, is suitable for children immunization. A parenteral cholera vaccine could overcome the diminishing immunogenicity in most of oral vaccines due to the gastrointestinal complexity and environmental enteropathy in children living in impoverished environment and could be considered for global cholera immunization.
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References
Chambers, J.S.: The conquest of Cholera: America’s Greatest Scourge. 1938. The Macmillan Publishing co. Inc., New York (1938)
Griffith, D.C., Kelly-Hope, L.A., Miller, M.A.: Review of reported cholera outbreaks worldwide, 1995–2005. Am. J. Trop. Med. Hyg. 75, 973–977 (2006)
Mutre, J., Kim, D.W., Thomson, N.R., Connor, T.R., Lee, J.H., Kariuki, S., Croucher, N.J., Choi, S.Y., Harris, S.R., Lebens, M., Niyogi, S.K., Kim, E.J., Ramamurthy, T., Chun, J., Wood, J.L., Clemens, J.D., Czerkinsky, C., Nair, G.B., Holmgren, J., Parkhill, J., Dougan, G.: Evidence for several waves of global transmission in the seventh cholera pandemic. Nature 477, 462–465 (2011)
Mintz, E.D., Guerrant, R.L.: A lion in our village – the unconscionable era of cholera in Africa. N. Engl. J. Med. 360, 1060–1063 (2009)
Bhattacharya, S., Black, R., Bourgeois, L., Clemens, J., Cravioto, A., Deen, J.L., Dougan, G., Glass, R., Grais, R.F., Greco, M., Gust, I., Holmgren, J., Kariuki, S., Lambert, P.H., Liu, M.A., Longini, I., Nair, G.B., Norrby, R., Nossal, G.J., Ogra, P., Sansonetti, P., von Seidlein, L., Songane, F., Svennerholm, A.M., Steele, D., Walker, R.: Cholera crises in Africa. Science 325, 885 (2009)
Chambers, K.: Zimbabwe’s battle against cholera. Lancet 373, 993–994 (2009)
Ali, M., Lopez, A.L., You, Y.A., Kim, Y.E., Sah, B., Maskery, B., Clemens, J.: The global burden of cholera. Bull. World Health Organ. 90, 209–218 (2012)
Leung, D.T., Chowdhury, F., Calderwood, S.B., Qadri, F., Ryan, E.T.: Immune responses to cholera in children. Expert Rev. Anti Infect. Ther. 10, 435–444 (2012)
Sekar, R., Amudhan, M., Sivashankar, M., Mythily, N., Mythreyee, M.A.: An outbreak of cholera among a rural population in south India: is it time to vaccinate the children in endemic areas? Indian J. Med. Res. 135, 678–679 (2012)
Benenson, A.S., Mosley, W.H., Fahimuddin, M., Oseasohn, R.O.: Cholera vaccine field trials in east Pakistan. 2. Effectiveness in the field. Bull. World Health Organ. 38, 359–372 (1968)
Barzilay, E.J., Schaad, N., Magloire, R., Mung, K.S., Boncy, J., Dahourou, G.A., Mintz, E.D., Steenland, M.W., Vertefeuille, J.F., Tappero, J.W.: Cholera surveillance during the Haiti epidemic–the first 2 years. N. Engl. J. Med. 368, 599–609 (2013)
Date, K.A., Vicari, A., Hyde, T.B., Mintz, E., Danovaro-Holliday, C., Henry, A., Tappero, J.W., Roels, T.H., Abrams, J., Burkholder, B.T., Ruiz-Matus, C., Andrus, J., Dietz, V.: Considerations for oral cholera vaccine use during outbreak after earthquake in Haiti. 2010–2011. Emerg. Infect. Dis. 17, 2105–2112 (2011)
Enserink, M.: Public health, no vaccines in the time of cholera. Science 329, 1462–1463 (2010)
Harris, J.B., LaRocque, R.C., Qadri, F., Ryan, E.T., Calderwood, S.B.: Cholera. Review. Lancet 379, 2466–2476 (2012)
GAVI Alliance.: Vaccine Investment Strategy Working Group Review. Washington DC 29–30 September, 2009. p. 2. http://www.gavialliance.org/library/gavi-documents/strategy/ (2009)
Mosley, W.H., Woodward, W.E., Aziz, K.M., Rahman, A.S., Chowdhury, A.K., Ahmed, A., Feeley, J.C.: The 1968–1969 cholera-vaccine field trial in rural East Pakistan. Effectiveness of monovalent Ogawa and Inaba vaccines and a purified Inaba antigen, with comparative results of serological and animal protection tests. J. Infect. Dis. Suppl. 121, 1–9 (1970)
Sommer, A., Khan, M., Mosley, W.H.: Efficacy of vaccination of family contacts of cholera cases. Lancet 1(7814), 1230–1232 (1973)
Sommer, A., Mosley, W.H.: Ineffectiveness of cholera vaccination as an epidemic control measure. Lancet 1(7814), 1232–1235 (1973)
Richie, E.E., Punjabi, N.H., Sidharta, Y.Y., Peetosutan, K.K., Sukandar, M.M., Wasserman, S.S., Lesmana, M.M., Wangsasaputra, F.F., Pandam, S.S., Levine, M.M., O’Hanley, P.P., Cryz, S.J., Simanjuntak, C.H.: Efficacy trial of single-dose live oral cholera vaccine CVD 103-HgR in North Jakarta, Indonesia, a cholera-endemic area. Vaccine 18, 2399–2410 (2000)
Levine, M.M.: Immunogenicity and efficacy of oral vaccines in developing countries: lessons from a live cholera vaccine. BMC Biol. 8, 129–139 (2010)
Clemens, J.D., van Loon, F., Sack, D.A., Chakraborty, J., Rao, M.R., Ahmed, F., Harris, J.R., Khan, M.R., Yunus, M., Huda, S., et al.: Field trial of oral cholera vaccines in Bangladesh: serum vibriocidal and antitoxic antibodies as markers of the risk of cholera. J. Infect. Dis. 163, 1235–1242 (1991)
Sur, D., Kanungo, S., Sah, B., Manna, B., Ali, M., Paisley, A.M., Niyogi, S.K., Park, J.K., Sarkar, B., Puri, M.K., Kim, D.R., Deen, J.L., Holmgren, J., Carbis, R., Rao, R., Nguyen, T.V., Han, S.H., Attridge, S., Donner, A., Ganguly, N.K., Bhattacharya, S.K., Nair, G.B., Clemens, J.D., Lopez, A.L.: Efficacy of a low-cost, inactivated whole-cell oral cholera vaccine: results from 3 years of follow-up of a randomized, controlled trial. PLoS Negl. Trop. Dis. 5, e1289 (2011)
Ahmed, A., Bhattacharjee, A.K., Mosley, W.H.: Characteristics of the serum vibriocidal and agglutinating antibodies in cholera cases and in normal residents of the endemic and non-endemic cholera areas. J. Immunol. 105, 432–441 (1970)
Levine, M.M., Nalin, D.R., Craig, J.P., Hoover, D., Bergquist, E.J., Waterman, D., et al.: Immunity of cholera in man: relative role of antibacterial versus antitoxic immunity. Trans. R. Soc. Trop. Med. Hyg. 73, 3–9 (1979)
Gupta, R.K., Szu, S.C., Finkelstein, R.A., Robbins, J.B.: Synthesis, characterization, and some immunological properties of conjugates composed of the detoxified lipopolysaccharide of Vibrio cholerae O1 serotype Inaba bound to cholera toxin. Infect. Immun. 60, 3201–3208 (1992)
Mosley, W.H., Benenson, A.S., Barui, R.: A serological survey for cholera antibodies in rural east Pakistan. 2. A comparison of antibody titres in the innunized and control populationd of a cholera-vaccine field-trial area and the relation of antibody titre to cholera case rate. Bull. World Health Organ. 38, 335–346 (1968)
McCormack, W.M., Chakraborty, J., Rahman, A.S., Mosley, W.H.: Vibriocidal antibody in clinical cholera. J. Infect. Dis. 120, 192–201 (1969)
Mosley WH, Ahmad S, Benenson AS, Ahmed A.: The relationship of vibriocidal antibody titre to susceptibility to cholera in family contacts of cholera patients. Bull. World Health Organ. 38:777–785
Chongsa-nguan, M., Chaicumpa, W., Kalambaheti, T., Soejoedi, H., Luxananil, P., Swasdikosa, S., Thanasiri, P., Mayurasakorn, S.: Vibriocidal antibody and antibodies to Vibrio cholerae lipopolysaccharide, cell-bound haemagglutinin and toxin in Thai population. Southeast Asian J. Trop. Med. Public Health 17, 558–566 (1986)
Neoh, S.H., Rowley, D.: The antigens of Vibrio cholerae involved in vibriocidal action of antibody and complement. J. Infect. Dis. 121, 505–513 (1970)
Patel, S.M., Rahman, M.A., Mohasin, M., Riyadh, M.A., Leung, D.T., Alam, M.M., Chowdhury, F., Khan, A.I., Weil, A.A., Aktar, A., Nazim, M., LaRocque, R.C., Ryan, E.T., Calderwood, S.B., Qadri, F., Harris, J.B.: Memory B cell responses to Vibrio cholerae O1 lipopolysaccharide are associated with protection against infection from household contacts of patients with cholera in Bangladesh. Clin. Vaccine Immunol. 19, 842–848 (2012)
Kossaczka, Z., Szu, S.C.: Evaluation of synthetic schemes to prepare immunogenic conjugates of Vibrio cholerae O139 capsular polysaccharide with chicken serum albumin. Glycoconj. J. 17, 425–433 (2000)
Gupta, R.K., Taylor, D.N., Bryla, D.A., Robbins, J.B., Szu, S.C.: Phase I evaluation of Vibrio cholerae O1, serotype Inaba, polysaccharide-cholera toxin conjugates in adult volunteers. Infect. Immun. 66, 3095–3099 (1998)
Kenne, L., Lindberg, B., Unger, P., Gustafsson, B., Holme, T.: Structural studies of the Vibrio cholerae O-antigen. Carbohydr. Res. 100, 341–349 (1982)
Hisatsune, K., Kondo, S., Isshiki, Y., Iguchi, T., Haishima, Y.: Occurrence of 2-O-ethyl-N-(3-deoxy-L-glycero-tetronyl)-D-perosamine (4-amino-4,6-dideoxy-D-anno-pyranose) in lipopolysaccharide from Ogawa but not from Inaba O forms of O1 Vibrio cholerae. Biochem. Biophys. Res. Commun. 190, 302–307 (1993)
Ito, T., Higuchi, T., Hirobe, M., Hiramatsu, K., Yokota, T.: Identification of a novel sugar, 4-amino-4,6,-dideoxy-o-methylmannose in the lipopolysaccharide of Vibrio cholerae O1 serotype Ogawa. Carbohydr. Res. 256, 113–128 (1998)
Zhang, J., Kovác, P.: Synthesis of methyl alpha-glycosides of some higher oligosaccharide fragments of the O-antigen of Vibrio cholerae O1, serotype Inaba and Ogawa. Carbohydr. Res. 300, 329–439 (1997)
Chernyak, A., Kondo, S., Wade, W.T., Meeks, M., Alzari, P., Fournier, J.-M.: Induction of protective immunity by synthetic Vibrio cholerae hexasaccharide derived from V. cholerae O1 Ogawa lipopolysaccharide bound to a protein carrier. J. Infect. Dis. 185, 950–962 (2002)
Meeks, M.D., Saksena, R., Ma, X., Wade, T.K., Taylor, R.K., Kovác, P., Wade, W.T.: Synthetic fragments of Vibrio cholerae O1 Inaba O-specific polysaccharide bound to a protein carrier are immunogenic in mice but do not induce protective antibodies. Infect. Immun. 72, 4090–4101 (2004)
Xu, P., Alam, M.M., Kalsy, A., Charles, R.C., Calderwood, S.B., Qadri, F., Ryan, E.T., Kováč, P.: Direct conjugation of bacterial O-SP-core antigens to proteins: development of cholera conjugate vaccines. Bioconjug. Chem. 22, 2179–2185 (2011)
Pozsgay, V., Chu, C., Pannell, L., Wolfe, J., Robbins, J.B., Schneerson, R.: Protein conjugates of synthetic saccharides elicit higher levels of serum IgG lipopolysaccharide antibodies in mice than do those of the O-specific polysaccharide Shigella dysenteriae type 1. Proc. Natl. Acad. Sci. U. S. A. 96, 5194–5197 (1999)
Safari, D., Dekker, H.A., de Jong, B., Rijkers, G.T., Kamerling, J.P., Snippe, H.: Antibody- and cell-mediated immune responses to a synthetic oligosaccharide conjugate vaccine after booster immunization. Vaccine 29, 6498–6504 (2011)
Costantino, P., Rappuoli, R., Berti, F.: The design of semi-synthetic and synthetic glycoconjugate vaccines. Expert. Opin. Drug Discov. 6, 1045–1066 (2011)
Pozsgay, V.: Recent developments in synthetic oligosaccharide-based bacterial vaccines. (Review). Curr. Top. Med. Chem. 8, 126–140 (2008)
Gening, M.L., Maira-Litran, T., Kropec, A., Shurnik, D., Grout, M., Tsvetkov, Y.E., Nifantiev, N.E., Pier, G.B.: Synthetic β-(1–6)-linked N-acetylated and nonacetylated oligoglucosamines used to produce conjugate vaccines for bacterial pathogens. Infect. Immun. 78, 764–772 (2010)
Kusama, H., Craig, J.P.: Production of biologically active substances by two strains of vibrio cholerae. Infect. Immun. 1, 80–87 (1970)
Holmes, R.K., Vasil, M.L., Finkelstein, R.A.: Studies on toxinogenesis in Vibrio cholerae. III. Characterization of nontoxinogenic mutants in vitro and in experimental animals. J. Clin. Invest. 55, 551–560 (1975)
Bundle, D.R., Gerken, M., Peters, T.: Synthesis of antigenic determinants of the Brucella A antigen, utilizing methyl 4-azido-4,6-dideoxy-α-D-mannopyranoside efficiently derived from D-mannose. Carbohydr. Res. 174, 239–251 (1988)
Eis, M.J., Ganem, B.: An improved synthesis of D-perosamine and some derivatives. Carbohydr. Res. 176, 316–323 (1988)
Peters, T., Bundle, D.R.: Synthetic antigenic determinants of the Brucella A polysaccharirde: A disaccharide thioglycoside for block synthesis of pentasaccharide and lower homologues of a1,2-linked 4,6-dideoxy-4-formamido-a-D-mannose. Can. J. Chem. 67, 491–496 (1989)
Kenne, L., Unger, P., Wehler, T.: Synthesis and nuclear magnetic resonance studies of some N-acelated methyl 4-amino-4,6-dideoxy-a-D-mannopyranosides. J. Chem. Soc. Perkin Trans. 1, 1183–1196 (1988)
Saksena, R., Zhang, J., Kovac, P.: Immunogens from a hexaccharide fragment of the O-SP of the Vibrio cholerae O:1, serotype Ogawa. Tetrahedron Assymetry 16, 187–197 (2005)
Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A., Smith, F.: A colorimetric method for the determination of sugars. Nature 168, 167 (1951)
Fekete A., Hoogerhout P., Zomer G., Kubler-Kielb J., Schneerson R., Robbins J.B., Pozsgay V.: Synthesis of octa- and dodecamers of D-ribitol-1-phosphate and their protein conjugates. Carbohydr. Res. 341, 2037–2048 (2006)
Finkelstein, R.A.: Vibriocidal antibody inhibition (VAI) analysis: a technique for the identification of the predominant vibriocidal antibodies in serum and for the detection and identification of Vibrio cholerae antigens. J. Immunol. 89, 264–267 (1962)
Attridge, S.R., Johansson, C., Trach, D.D., Qadri, F., Svennerholm, A.M.: Sensitive microplate assay for detection of bactericidal antibodies to Vibrio cholerae O139. Clin. Diagn. Lab. Immunol. 9, 383–387 (2002)
Boutonnier, A., Dassy, B., Duménil, R., Guénolé, A., Ratsitorahina, M., Migliani, R., Fournier, J.M.: A simple and convenient microtiter plate assay for the detection of bactericidal antibodies to Vibrio cholerae O1 and Vibrio cholerae O139. J. Microb. Methods 55, 745–753 (2003)
Chu, C.Y., Liu, B.K., Watson, D., Szu, S.S., Bryla, D., Shiloach, J., Schneerson, R., Robbins, J.: Preparation, characterization, and immunogenicity of conjugates composed of the O-specific polysaccharide of Shigella dysenteriae type 1 (Shiga’s bacillus) bound to tetanus toxoid. Infect. Immun. 59, 4450–4458 (1991)
Mosley, W.H.: The role of immunity in cholera. A review of epidemiological and serological studies. Tex. Rep. Biol. Med. 27(Suppl.1), 227–241 (1969)
Sirisinha, S., Charupatana, C.: Antibody responses in serum, secretions, and urine of man after parenteral administration of vaccines. Infect. Immun. 2, 29–37 (1970)
Robbins, J., Schneerson, R., Szu, S.C.: Perspective: Hypothesis: Serum IgG antibody is sufficient to confer protection against infectious diseases by inactivating the innoculum. J. Infect. Dis. 171, 1387–1398 (1995)
Huang, W., Morrell, D.: Successful treatment of recalcitrant warts with topical squaric acid in immunosuppressed child. Pediatr. Dermatol. 25, 275–276 (2008)
Hou, S.J., Saksena, R., Kovác, P.: Preparation of glycoconjugates by dialkyl squarate chemistry revisited. Carbohydr. Res. 343, 196–210 (2008)
Saksena, R., Ma, X., Wade, T.K., Kovác, P., Wade, W.F.: Length of the linker and the interval between immunizations influences the efficacy of Vibrio cholerae O1, Ogawa hexasaccharide neoglycoconjugates. FEMS Immunol. Med. Microbiol. 47, 116–128 (2006)
Saksena, R., Ma, X., Wade, T.K., Kovác, P., Wade, W.F.: Effect of saccharide length on the immunogenicity of neoglycoconjugates from synthetic fragments of the O-SP of Vibrio cholerae O1, serotype Ogawa. Carbohydr. Res. 340, 2256–2269 (2005)
Finkelstein, R.A., Pongpairojana, S.: A test of antigenicity for the selection of strains for inclusion in cholera vaccines. Bull. World Health Organ. 39, 247–259 (1968)
Villeneuve, S., Boutonnier, A., Mulard, L.A., Fourier, J.-M.: Immunochemical characterization of an Ogawa-Inaba common antigenic determinant of Vibrio cholerae O1. Microbiology 145, 2477–2484 (1999)
Newman, B.A., Kabat, E.A.: An immunochemical study of the combining site specificities of C57BL/6J monoclonal antibodies to alpha (1–6)-linked dextran B512. J. Immunol. 135, 1220–1231 (1985)
Pozsgay, V., Kubler-Kielb, J., Schneerson, R., Robbins, J.B.: Effect of the nonreducing end of Shigella dysenteriae type 1 O-specific oligosaccharides on their immunogenicity as conjugates in mice. Proc. Natl. Acad. Sci. U. S. A. 104, 14478–14482 (2007)
World Health Organization: Zimbabwe Cholera and health situation. http://www.who.int/hac/crises/zmb/appeal/zimbabwe_cholera_advocacy_1dec (2008)
Mandal, J., Dinoop, K.P., Parija, S.C.: Increasing antimicrobial resistance of Vibrio cholerae OI biotype E1 tor strains isolated in a tertiary-care centre in India. J. Health Popul. Nutr. 30, 12–16 (2012)
Armah, G.E., Sow, S.O., Breiman, R.F., Dallas, M.J., Tapia, M.D., Feikin, D.R., Binka, F.N., Steele, A.D., Laserson, K.F., Ansah, N.A., Levine, M.M., Lewis, K., Coia, M.L., Attah-Poku, M., Ojwando, J., Rivers, S.B., Victor, J.C., Nyambane, G., Hodgson, A., Schödel, F., Ciarlet, M., Neuzil, K.M.: Efficacy of pentavalent rotavirus vaccine against severe rotavirus gastroenteritis in infants in developing countries in sub-Saharan Africa: a randomised, double-blind, placebo-controlled trial. Lancet 376, 606–614 (2010)
Acknowledgments
The authors would like to thank Drs. Nancy Vieira, Bruce Coxon, Jianping Li and Joanna Kubler-Kielb for to their helps in spectrum analysis and to Drs. John B. Robbins for helpful discussions.
Funding and disclosure
This work was supported fully by the Intramural Research at the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health. The authors have no conflict interest to disclose.
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Ftacek, P., Nelson, V. & Szu, S.C. Immunochemical characterization of synthetic hexa-, octa- and decasaccharide conjugate vaccines for Vibrio cholerae O:1 Serotype Ogawa with emphasis on antigenic density and chain length. Glycoconj J 30, 871–880 (2013). https://doi.org/10.1007/s10719-013-9491-9
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DOI: https://doi.org/10.1007/s10719-013-9491-9