- Research Article
The immunogenicity of the liposome-associated outer membrane proteins (OMPs) of Moraxella catarrhalis
Cellular & Molecular Biology Letters volume 15, Article number: 70 (2010)
The outer membrane proteins (OMPs) are the most immunogenic and attractive of the Moraxella catarrhalis vaccine antigens that may induce the protective immune response. The aim of this study was to determine the effectiveness of two types of OMP-associated phosphatidylcholine (PC) liposomal formulations (OMPs-PC, PC-OMPs) and of Zwittergent-based proteomicelles (OMPs-Z) in potentiating an anti-OMP systemic immune response in mice. The immunogenicities of the above preparations were evaluated by assessing serum anti-OMP IgG and IgA reactivity in the post-immunized mouse antisera using ELISA and Western blotting. Additionally, the cross-reactivity of the most effective anti-OMP response was determined using heterologous sera from both humans and mice. Both the proteoliposomes and the proteomicelles showed high immunogenic properties and did not elicit any distinct quantitative differences in the antibody titer or qualitative differences in the pattern of the mouse antisera. The post-immunized mouse antisera predominantly recognized a ∼60-kDa OMP of M. catarrhalis. That protein was also found to be a highly cross-reactive antigen interacting with a panel of pooled mouse antisera produced by immunization either with whole cells or the purified OMPs of heterologous M. catarrhalis strains. Furthermore, normal sera collected from healthy children were found to be preferentially reactive with the 60-kDa OMP. The serum-specific IgG, IgA and IgM were respectively detected via immunoblotting in 90%, 85% and 30% of heterologous human sera. This similar immunogenic effectiveness of both OMP-associated liposomal formulations could contribute to the practical use of such formulations in the future in human vaccination. Moreover, the highly cross-reactive 60-kDa OMP seems to be an important antigenic marker of M. catarrhalis, and, as it is responsible for the induction of an antibody-mediated and long-lasting immune response, studying it may partially aid us in understanding the relatively low degree of pathogenicity of the bacterium in immunocompetent individuals.
atomic force microscopy
bovine serum albumin
enzyme-linked immunosorbent assay
outer membrane proteins
- OMPs-PC and PC-OMPs:
the studied proteoliposomes
Zwittergent-based micelles of OMPs
phosphate buffered saline
PBS supplemented with Tween 20
Karaulus, R. and Campagnari, A. Moraxella catarrhalis: a review of an important human mucosal pathogen. Microb. Infect. 2 (2000) 547–559.
Murphy, T.F., Brauer, A.L., Aebi, Ch. and Sethi, S. Identification of surface antigens of Moraxella catarrhalis as targets of human serum antibody responses in chronic obstructive pulmonary disease. Infect. Immun. 73 (2005) 3471–3478.
Murphy, T.F., Brauer, A.L., Grant, B.J. and Sethi, S. Moraxella catarrhalis in chronic obstructive pulmonary disease: burden of disease and immune response. Am. J. Respir. Crit. Care. Med. 172 (2005) 195–199.
Sethi, S., Sethi, R., Eschberger, K., Lobbins, P., Cai, X., Grant, B.J. and Murphy, T.F. Airway bacterial concentrations and exacerbations of chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 176 (2007) 356–361.
Verduin, C.M. Hol, C. Fleer, A. van Dijk, H. and van Belkum, A. Moraxella catarrhalis: from emerging to established pathogen. Clin. Microb. Rev. 15 (2002) 125–144.
Bandak, S.M., Turnak, M.R., Allen, B.S., Bolzon, L.D., Preston, D.A., Bouchillon, S.K. and Hoban, D.J. Antibiotic susceptibility among recent clinical isolates of Haemophilus influenza and Moraxella catarrhalis from fifteen countries. Eur. J. Clin. Microbiol. Infect. Dis. 20 (2001) 55–60.
Klingman, K.L., Pye, A., Murphy, T.F. and Hill, S.L. Dynamics of respiratory tract colonization by Branhamella catarrhalis in bronchiectasis. Am. J. Respir. Crit. Care. Med. 152 (1995) 1072–1078.
Faden, H., Duffy, L., Wasielewski, R., Wolf, J., Krystofik, D. and Tung, Y. Relationship between nasopharyngeal colonization and the development of otitis media in children. J. Infect. Dis. 175 (1997) 1440–1445.
Yokota, S., Harimaya, A., Sato, K., Somekawa, Y., Himi, T. and Fujii, N. Colonization and turnover of Streptococcus pneumonia, Haemophilus influenzae, and Moraxella catarrhalis in otitis-prone children. Microbiol. Immunol. 51 (2007) 223–230.
Heiniger, N., Spaniol, V., Troller, R., Vischer, M. and Aebi, C. A reservoir of Moraxella catarrhalis in human pharyngeal lymphoid tissue. J. Infect. Dis. 196 (2007) 1080–1087.
Peng, D., Hong, W., Choudhury, B., Carlson, R.W. and Gu, X.X. Moraxella catarrhalis bacterium without endotoxin, a potential vaccine candidate. Infect. Immun. 73 (2005) 7569–7577.
Troncoso, G., Sánchez, S., Criado, M.T. and Ferreirós, C. Analysis of Moraxella catarrhalis outer membrane antigens cross-reactive with Neisseria meningitidis and Neisseria lactamica. FEMS Immunol. Med. Microbiol. 40 (2004) 89–94.
Mleczko, J., Augustyniak, D. and Jankowski, A. Efficiency of oral immunization of mice with Candida albicans and Moraxella catarrhalis heat-killed cells and cross reactivity of induced antibodies. Centr. Eur. J. Immunol. 32 (2007) 185–188.
Steeghs, L., Kuipers, B., Hamstra, H.J., Kersten, G., Van Alphen, L. and Van der Ley, P. Immunogenicity of outer membrane proteins in a lipopolysaccharide —deficient mutant of Neisseria meningitidis: influence of adjuvants on the immune response. Infect. Immun. 67 (1999) 4988–4993.
Schweizer, M., Hindennach, I., Garten, W. and Henning, U. Major proteins of the Escherichia coli outer cell envelope membrane. Interaction of protein II* with lipopolysaccharide. Eur. J. Biochem. 82 (1978) 211–217.
Bogdanov, M. and Dowhan, W. Lipid-assisted protein folding. J. Biol. Chem. 274 (1999) 36827–36830.
Qi, H.L., Tai, J.Y. and Blake, M.S. Expression of large amounts of neisserial porin proteins in Escherichia coli and refolding of the proteins into native trimers. Infect. Immun. 62 (1994) 2432–2439.
Wetzler, L.M., Blake, M.S. and Gotschlich, E.C. Characterization and specificity of antibodies to protein I of Neisseria gonorrhoeae produced by injection with various protein I-adjuvant preparation. J. Exp. Med. 168 (1988) 1883–1897.
Jolley, K.A., Appleby, L., Wright, J.C., Christodoulides, M. and Heckels, J. Immunization with recombinant Opc outer membrane protein from Neisseria meningitidis: Influence of sequence variation and levels of expression on the bactericidal immune response against meningococci. Infect. Immun. 69 (2001) 3809–3916.
Frézard, F. Liposomes: from biophysics to the design of peptide vaccines. Braz. J. Med. Biol. Res. 32 (1999)181–189.
Altin, J.G. and Parish, Ch.R. Liposomal vaccines-targeting the delivery of antigen. Methods 40 (2006) 39–52.
Uchida, T. and Taneichi, M. Clinical application of surface-linked liposomal antigens. Mini Rev. Med. Chem. 8 (2008) 184–192.
Parmar, M.M., Edwards, K. and Madden, T.D. Incorporation of bacterial membrane proteins into liposomes: factors influencing protein reconstitution. Biochim. Biophys. Acta 1421 (1999) 77–90.
Stebelska, K., Wyrozumska, P., Gubernator, J. and Sikorski A.F. Highly fusogenic cationic liposomes transiently permeabilize the plasma membrane of HeLa cells. Cell. Mol. Biol. Lett. 12 (2007) 35–39.
Gutowicz, J. and Terlecki, G. The association of glycolytic enzymes with cellular and model membranes. Cell. Mol. Biol. Lett. 8 (2003) 667–680.
Murphy, T.F. and Bartos, L.C. Surface-exposed and antigenically conserved determinants of outer membrane proteins of Branhamella catarrhalis. Infect. Immun. 57 (1989) 2938–2941.
Rigaud, J-L. and Lévy, D. Reconstitution of membrane proteins into liposomes. in: Methods in Enzymol. Liposomes, Part B. 372 Elsevier Academic Press, Co. Elseviere Inc., 2003,65–86.
Rigaud, J.L., Paternoster, M.T. and Bluza, A. Mechanisms of membrane protein insertion into liposomes during reconstitution procedures involving the use of detergents.2. Incorporation of the light-driven proton pump bacteriorhodopsin. Biochemistry 27 (1988) 2677–2688.
Sarwar, J., Campagnari, A.A., Kirkham, C. and Murphy, T. Characterization of an antigenically conserved heat-modifiable major outer membrane protein of Branhamella catarrhalis. Infect. Immun. 60 (1992) 804–809.
Mandrell, R.E. and Zollinger, W.D. Use of a zwitterionic detergent for the restoration of the antibody-binding capacity of electroblotted meningococcal outer membrane proteins. J. Immunol. Met. 67 (1984) 1–11.
Idänpään-Heikkilä, I., Wahlström, E., Muttilainen, S., Nurminen, M., Käyhty, H., Sarvas, M. and Mäkelä, P.H. Immunization with meningococcal class 1 outer membrane protein produced in Bacillus subtilis and reconstituted in the presence of Zwittergent or Triton X-100. Vaccine 14 (1996) 886–891.
Shahum, E. and Thérien, H.M. Effect of liposomal antigens on the priming and activation of the immune system by dendritic cells. Int. Immunopharmacol. 2 (2002) 591–601.
Ignatius, R., Mahnke, K., Rivera, M., Hong, K., Isdell, F., Steinman, R.M., Pope, M. and Stamatatos, L. Presentation of proteins encapsulated in sterically stabilized lioposomes by dendritic cells initiates CD8(+) T-cell response in vivo. Blood 96 (2000) 3505–3513.
Alving, C.R. Immunologic aspects of liposomes: presentation and processing of liposomal protein and phospholipid antigens. Biochim. Biophys. Acta 1113 (1992) 307–322.
Wright, J.C., Wiliams, J.N., Christodoulides, M. and Haeckels, J.E. Immunization with the recombinant PorB membrane prorein induces bactericidal immune response against Neisseria meningitidis. Infect. Immune. 70 (2002) 4028–4034.
Sprott, G.D., Dicaire, C.J., Gurnani, K., Deschatelets, L.A. and Krishnan, L. Liposome adjuvants prepared from the total polar lipids of Haloferax volcanii, Planococcus spp. and Bacillus firmus differ in ability to elicit and sustain immune responses. Vaccine 22 (2004) 2154–2162.
Witkowska, D., Masłowska, E., Staniszewska, M., Szosto, B., Jankowski, A. and Gamian, A. Enterobacterial 38-kDa outer membrane protein is an age-dependent molecular marker of innate immunity and immunoglobulin deficiency as results from its reactivity with IgG and IgA antibody. FEMS Immunol. Med. Microbiol. 48 (2006) 205–214.
Christiensen, J.J. Moraxella (Branhamella) catarrhalis: clinical, microbiological and immunological features in lower respiratory tract infections. APMIS 107S (1999) 1–36.
Mathers, K., Leinonen, M. and Goldblatt, D. Anibody response to outer membrane proteins of Moraxella catarrhalis in children with otitis media. Pediatr. Infect. Dis. J. 18 (1999) 982–988.
Murphy, T.F., Kirkham, C., Liu, D.F. and Sethi, S. Human immune response to outer membrane protein CD of Moraxella catarrhalis in adults with chronic obstructive pulmonary disease. Infect. Immun. 71 (2003) 1288–1294.
Murphy, T.F., Kirkham, C. and Lesse, A.J. The major heat modifiable outer membrane protein CD is highly conserved among strains of Branhamella catarrhalis. Mol. Microbiol. 10 (1993) 87–97.
Hsiao, C.B., Sethi, S. and Murphy, T.F. Outer membrane protein CD of Branhamella catarrhalis: sequence conservation in strains recovered from the human respiratory tract. Microb. Pathog. 19 (1995) 215–225.
McMichael, J. Vaccines for Moraxella catarrhalis. Vaccine 19 (2001) 101–107.
Yang-Ping, Y., Myers, L.E., McGuinnes, U., Chong, P., Kwok, Y., Klein, M.H. and Harkness, R.E. The major outer membrane protein, CD, extracted from Moraxella (Branhamella) catarrhalis is a potential vaccine antigen that induces bactericidal antibodies. FEMS Immunol. Med. Microbiol. 17 (1997) 187–199.
Meier, P.S., Freiburghaus, S., Martin, A., Heiniger, N., Troller, R. and Aebi, C. Mucosal immune response to specific outer membrane proteins of Moraxella catarrhalis in young children. Pediatr. Infect. Dis. J. 22 (2003) 256–262.
Meier, P.S., Heiniger, N., Troller, R. and Aebi, C. Salivary antibodies directed against outer membrane proteins of Moraxella catarrhalis in healthy adults. Infect. Immun. 71 (2003) 6793–6798.
Holm, M.M., Vanlerberg, S.L., Foley, I.M., Sledjeski, D.D. and Lafontaine, E.R. The Moraxella catarrhalis porin-like outer membrane protein CD is an adhesion for human lung cells. Infect. Immun. 72 (2004)1906–1913.
Liu, D.F., McMichael, J.C. and Baker, S.M. Moraxella catarrhalis outer membrane protein CD elicits antibodies that inhibit CD binding to human mucin and enhance pulmonary clearance of M.catarrhalis in a mouse model. Infect. Immun. 75 (2007) 2818–2825.
Hu, W.G., Berry, J., Chen, J. and Gum, X-X. Exploration of Moraxella catarrhalis outer membrane proteins, CD and UspA, as new carriers for lipooligosaccharide-based conjugates. FEMS Immunol. Med. Microbiol. 41 (2004)109–115.
About this article
Cite this article
Augustyniak, D., Mleczko, J. & Gutowicz, J. The immunogenicity of the liposome-associated outer membrane proteins (OMPs) of Moraxella catarrhalis . Cell Mol Biol Lett 15, 70 (2010). https://doi.org/10.2478/s11658-009-0035-z
- Moraxella catarrhalis
- Outer membrane proteins
- Anti-OMP antibodies