Our research on pathogenesis of disseminated candidiasis led to the discovery that antibodies specific for Candida albicans cell surface β-1, 2–mannotriose [β-(Man)3] protect mice. A 14 mer peptide Fba, which derived from the N-terminal portion of the C. albicans cytosolic/cell surface protein fructose-bisphosphate aldolase, was used as the glycan carrier and resulted in a novel synthetic glycopeptide vaccine β-(Man)3-Fba. By a dendritic cell-based immunization approach, this conjugate induced protective antibody responses against both the glycan and peptide parts of the vaccine. In this report, we modified the β-(Man)3-Fba conjugate by coupling it to tetanus toxoid (TT) in order to improve immunogenicity and allow for use of an adjuvant suitable for human use. By new immunization procedures entirely compatible with human use, the modified β-(Man)3-Fba-TT was administered either alone or as a mixture made with alum or monophosphoryl lipid A (MPL) adjuvants and given to mice by a subcutaneous (s.c.) route. Mice vaccinated with or, surprisingly, without adjuvant responded well by making robust antibody responses. The immunized groups showed a high degree of protection against a lethal challenge with C. albicans as evidenced by increased survival times and reduced kidney fungal burden as compared to control groups that received only adjuvant or DPBS buffer prior to challenge. To confirm that induced antibodies were protective, sera from mice immunized against the β-(Man)3-Fba-TT conjugate transferred protection against disseminated candidiasis to na？ve mice, whereas C. albicans-absorbed immune sera did not. Similar antibody responses and protection induced by the β-(Man)3-Fba-TT vaccine was observed in inbred BALB/c and outbred Swiss Webster mice. We conclude that addition of TT to the glycopeptide conjugate results in a self-adjuvanting vaccine that promotes robust antibody responses without the need for additional adjuvant, which is novel and represents a major step forward in vaccine design against disseminated candidiasis.
References
[1]
Hsu FC, Lin PC, Chi CY, Ho MW, Wang JH (2009) Prognostic factors for patients with culture-positive Candida infection undergoing abdominal surgery. J Microbiol Immunol Infect 42: 378–384.
[2]
Horn DL, Neofytos D, Anaissie EJ, Fishman JA, Steinbach WJ, et al. (2009) Epidemiology and outcomes of candidemia in 2019 patients: data from the prospective antifungal therapy alliance registry. Clin Infect Dis 48: 1695–1703.
[3]
Zaoutis TE, Argon J, Chu J, Berlin JA, Walsh TJ, et al. (2005) The epdiemiology and attributable outcomes of candidemia in adults and children hospitalized in the United States: A propensity analysis. Clin Infect Dis 41: 1232–1239.
[4]
Mochon AB, Cutler JE (2005) Is a vaccine needed against Candida albicans? Med Mycol 43: 97–115.
[5]
Cassone A, De Bernardis F, Torosantucci A (2005) An outline of the role of anti-Candida antibodies within the context of passive immunization and protection from candidiasis. Curr Mol Med 5: 377–382.
[6]
Cutler JE, Deepe GS Jr, Klein BS (2007) Advances in combating fungal diseases: Vaccines on the threshold. Nat Rev Microbiol 5: 13–28.
[7]
Xin H, Dziadek S, Bundle DR, Cutler JE (2008) Synthetic glycopeptide vaccines combining β-mannan and peptide epitopes induce protection against candidiasis. Proc Natl Acad Sci USA 105: 13526–13531.
[8]
Cutler JE (2005) Defining criteria for anti-mannan antibodies to protect against candidiasis. Curr Mol Med 5: 383–392.
[9]
De Bernardis F, Boccanera M, Adriani D, Spreghini E, Santoni G, et al. (1997) Protective role of antimannan and anti-aspartyl proteinase antibodies in an experimental model of Candida albicans vaginitis in rats. Infect Immun 65: 3399–3405.
[10]
Matthews RC, Rigg G, Hodgetts S, Carter T, Chapman C, et al. (2003) Preclinical assessment of the efficacy of mycograb, a human recombinant antibody against fungal HSP90. Antimicrob Agents Chemother 47: 2208–2216.
[11]
Yang Q, Wang L, Lu DN, Gao RJ, Song JN, et al. (2005) Prophylactic vaccination with phage-displayed epitope of C. albicans elicits protective immune responses against systemic candidiasis in C57Bl/6 mice. Vaccine 23: 4088–4096.
[12]
Cassone A, Rappuol R (2010) Universal vaccines: shifting to one for many. mBio 1: e00042–10.
[13]
Bromuro C, Romano M, Chiani P, Berti F, Tontini M, et al. (2010) Beta-glucan-CRM 197 conjugates as candidates antifungal vaccines. Vaccine 28: 2615–2623.
[14]
Krcmery V, Barnes AJ (2002) Non-albicans Candida spp. causing fungaemia: pathogenicity and antifungal resistance. J Hosp Infect 50: 243–260.
[15]
Perlroth J, Cho B, Spellberg B (2007) Nosocomial fungal infections: epidemiology, diagnosis, and treatment. Med Mycol 45: 321–346.
[16]
Cutler JE, Corti M, Lambert P, Ferris M, Xin H (2011) Horizontal transmission of Candida albicans and evidence of a vaccine response in mice colonized with the fungus. PLoS One 6: e22030.
[17]
Xin H, Cutler JE (2011) Vaccine and Monoclonal Antibody That Enhance Mouse Resistance to Candidiasis. Clin Vaccine Immunol 18: 1656–1667.
[18]
Han Y, Kozel TR, Zhang MX, MacGill RS, Carroll MC, et al. (2001) Complement is essential for protection by an IgM and an IgG3 monoclonal antibody against experimental hematogenously disseminated candidiasis. J Immunol 167: 1550–1557.
[19]
Xin H, Cutler JE (2006) Hybricoma passage in vitro may result in reduced ability of antimannan antibody to protect against disseminated candidiasis. Infect Immun 74: 4310–4321.
[20]
Kuroda E, Sugiura T, Zeki K, Yoshida Y, Yamashita U (2000) Sensitivity difference to the suppressive effect of prostaglandin E2 among mouse strains: a possible mechanism to polarize Th2 type response in BALB/c mice. J Immunol 1164: 2386–2395.
[21]
Ashman RB (1990) Murine candidiasis: Susceptibility is associated with the induction of T cell-mediated, strain-specific autoreactivity. Immunol Cell Biol 68: 179–185.
[22]
Ashman RB (1997) Genetic determination of susceptibility and resistance in the pathogenesis of Candida albicans infection. FEMS Microbiology Letters 19: 183–189.
[23]
Hector RF, Domer JE, Carrow EW (1982) Immune responses to Candida albicans in genetically distinct mice. Infect Immun 38: 1020–1028.
[24]
Didierlaurent A, Morel S, Lockman L, Giannini SL, Bisteau M, et al. (2009) AS04, an aluminum salt- and TLR4 agonist-based adjuvant system, induces a transient localized innate immune response leading to enhanced adaptive immunity. J Immunol 183: 6186–6197.
[25]
Han Y, Kanbe T, Cherniak R, Cutler JE (1997) Biochemical characterization of Candida albicans epitopes that can elicit protective and nonprotective antibodies. Infect Immun 65: 4100–4107.
[26]
Nitz M, Ling C-C, Otter A, Cutler JE, Bundle DR (2002) The unique solution structure and immunochemistry of the Candida albicans β-1,2-mannopyranan cell wall antiegns. J Biol Chem 277: 3440–3446.
[27]
Casadevall A, Pirofski L-A (2007) Antibody-mediated protection through cross-reactivity introduces a fungal heresy into immunological dogma. Infect Immun 75: 5074–5078.
[28]
Casadevall A, Dadachova E, Pirofski L-A (2004) Passive antibody therapy for infectious diseases. Nature Reviews 2: 695–703.
[29]
Han Y, Cutler JE (1995) Antibody response that protects against disseminated candidiasis. Infect Immun 63: 2714–2719.
[30]
Han Y, Morrison RP, Cutler JE (1998) A vaccine and monoclonal antibodies that enhance mouse resistance to Candida albicans vaginal infection. Infect Immun 66: 5771–5776.
[31]
Moyle PM, Toth I (2008) Self-adjuvanting lipopeptide vaccines. Curr Med Chem 15: 506–516.
[32]
Brown LE, Jackson DC (2005) Lipid-based Self-Adjuvanting Vaccines. Current Drug Delivery 2: 383–393.
[33]
Fotin-Mleczek M, Duchardt KM, Lorenz C, Pfeiffer R, Ojkic-Zrna S, et al. (2011) Messenger RNA-based vaccines with dual activity induce balanced TLR-7 dependent adaptive immune responses and provide antitumor activity. J Immunother 34: 1–15.
[34]
Maittra RW, Datta K, Lees A, Belouski SS, Pirofski L-A (2004) Immunogenicity and efficacy of Cryptococcus neoformans capsular polysaccharide glucuronoxylomannan peptide mimotope-protein conjugates in human immunoglobulin transgenic mice. Infect Immun 72: 196–208.
[35]
Germann T, Bongartz M, Dlugonska H, Hess H, Schmitt E, et al. (1995) Interleukin-12 profoundly up-regulates the synthesis of antigen-specific complement-fixing IgG2a, IgG2b and IgG3 antibody subclasses in vivo. Eur J Immunol 25: 823–829.
[36]
Nussbaum G, Yuan R, Casadevall A, Scharff MD (1996) Immunoglobulin G3 blocking antibodies to the fungal pathogen Cryptococcus neoformans. J Exp Med 183: 1905–1909.
[37]
Schreiber JR, Cooper LJN, Diehn S, Dahlhauser PA, Tosi MF, et al. (1993) Variable region-identical monoclonal antibodies of different IgG subclass directed to Pseudomonas aeruginosa lipopolysaccharide O-specific side chain fuction differently. JID 167: 221–226.
[38]
Cooper LJN, Schimenti JC, Glass DD, Greenspan NS (1991) H chain C domains influence the strength of binding of IgG for streptococcal group A carbohydrate. J Immunol 146: 2659–2663.
[39]
Unkeless JC, Sciqliano E, Freedman VH (1988) Structure and function of human and murine receptors for IgG. Annu Rev Immunol 251–281.
[40]
Han Y, Riesselman MH, Cutler JE (2000) Protection against candidiasis by an immunoglobulin G3 (IgG3) monoclonal antibody specific for the same mannotriose as an IgM protective antibody. Infect Immun 68: 1649–1654.
[41]
Han Y, Cutler JE (1997) Assessment of a mouse model of neutropenia and the effect of an anti-candidiasis monoclonal antibody in these animals. J Infect Dis 175: 1169–1175.
[42]
Wu X, Bundle DR (2005) Synthesis of glycoconjugate vaccines for Candida albicans using novel linker methodology. J Org Chem 70: 7381–7388.
[43]
Wu X, Lipinski T, Carrel FR, Bailey JJ, Bundle DR (2007) Synthesis and immunochemical studies on a Candida albicans cluster glycoconjugate vaccine. Org Biomol Chem 5: 3477–3485.
[44]
Kobayashi H, Matsuda K, Ikeda T, Suzuki M, Takahashi S, et al. (1994) Structures of cell wall mannans of pathogenic Candida tropicalis IFO 0199 and IFO 1647 yeast strains. Infect Immun 62: 615–622.
[45]
Shibata N, Kobayashi H, Okawa Y, Suzuki S (2003) Existence of novel β-1,2 linkage-containing side chain in the mannan of Candida lusitaniae, antigenically related to Candida albicans. Eur J Biochem 270: 2565–2575.
[46]
Shibata N, Akagi R, Hosoya T, Kawahara K, Suzuki A, et al. (1996) Existence of novel branched side chains containing β-1,2 and α-1,6 linkages corresponding to antigenic factor 9 in the mannan of Candida guilliermondii. Journal of Biological Chemistry 271: 9259–9266.
[47]
Suzuki A, Shibata N, Suzuki M, Saitoh F, Oyamada H, et al. (1997) Characterization of β-1,2-mannosyltransferase in Candida guilliermondii and its utilization in the synthesis of novel oligosaccharides. J Biol Chem 272: 16822–16828.
[48]
Goins T, Cutler JE (2000) Relative abundance of oligosaccharides in Candida species as determined by fluorophore-assisted carbohydrate electrophoresis. J Clin Microbiol 38: 2862–2869.
[49]
Kobayashi H, Mitobe H, Takahashi K, Yamamoto T, Shibata N, et al. (1992) Structural study of a cell wall mannan-protein complex of the pathogenic yeast Candida glabrata IFO 0622 strain. Arch Biochem Biophys 294: 662–669.
[50]
Dziadek S, Jacques S, Bundle DR (2008) A novel linker methodology for the synthesis of tailored conjugate vaccines composed of complex carbohydrate antigens and specific TH-cell peptide epitopes. Chemistry 14: 5908–5917.
[51]
Kanbe T, Cutler JE (1994) Evidence for adhesin activity in the acid-stable moiety of the phosphomannoprotein cell wall complex of Candida albicans. Infect Immun 62: 1662–1668.
[52]
Pincus SH, Shigeoka AO, Moe AA, Ewing LP, Hill HR (1988) Protective efficacy of IgM monoclonal antibodies in experimental group B streptococcal infection is a function of antibody. J Immunol 140: 2779–2785.
