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  • Applied Microbiology
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Measurement of opsonophagocytic activity of antibodies specific toNeisseria meningitidis serogroup A capsular polysaccharide-serogroup B outer membrane vesicle conjugate in animal model

Annals of Microbiology volume 59, pages 801–806 (2009)Cite this article

Abstract

Neisseria meningitidis is efficiently phagocytosed by polymorphonuclear leukocytes (PMNS) following opsonization with opsonic antibodies; opsonophagocytosis is the primary mechanism for clearance of meningococci from the host. Thus, in testing meningococcal vaccines, the level of opsonophagocytic antibodies appears to correlate with vaccine-induced protection. Our previous studies demonstrated that the conjugation ofN. meningitidis serogroup A capsular polysaccharide (CPSA) to serogroup B outer membrane vesicle (OMV) could induce a high level of bactericidal antibody response against serogroup A meningococci in animals. The purpose of this study was to evaluate opsonophagocytic activity of the conjugate of CPSA to OMV (CPSA-OMV). In order to evaluate the potential efficacy of CPSA-OMV a flow cytometric opsonophagocytic assay was used. The conjugate and controls were injected intramuscularly into four groups of rabbits with boosters on days 14, 28 and 42 following primary immunization. The rabbits were bled prior to injection and two weeks after each injection. Opsonophagocytic activity of antibodies in hyperimmune sera through rabbit PMNS were measured with flow cytometer, using dihydrorhodamine-123 as a probe. The results indicated that our conjugate could induce a highly significant level of opsonophagocytic activity against serogroup A meningococci after 56 days compared to the control groups (P<0.05). We conclude that this conjugate represents a vaccine candidate against serogroups A and B meningococci after further investigation.

References

  • Aase A., Bjune G., Hoiby E.A., Rosenqvist E., Pedersen A.K., Michaelsen T.E. (1995). Comparison among opsonic activity, antimeningococcal immunoglobulin G response, and serum. bactericidal activity against meningococci in sera from vaccinees after immunization with a serogroup B outer membrane vesicle vaccine. Infect. Immun., 63 (9): 3531–3536.

    CAS  PubMed  Google Scholar 

  • Aase A., Hoiby E.A., Michaelsen T.E. (1998). Opsonophagocytic and bactericidal activity mediated by purified IgG subclass antibodies after vaccination with the Norwegian group B meningococcal vaccine. Scand. J. Immunol., 47 (4): 388–396.

    Article  CAS  PubMed  Google Scholar 

  • Ada G., Isaacs D. (2003). Carbohydrate-protein conjugate vaccines. Clin. Microbiol. Infect., 9 (2): 79–85.

    Article  CAS  PubMed  Google Scholar 

  • Balmer P., Borrow R. (2004). Serologic correlates of protection for evaluating the response to meningococcal vaccines. Expert. Rev. Vaccines, 3 (1): 77–87.

    Article  CAS  PubMed  Google Scholar 

  • Behzadiyannejad Q., Siadat S.D., Kheirandish M., Tabaraie B., Ahmadi H., Norouzian D.,et al. (2008). Comparison among opsonic activity and serum bactericidal activity against meningococci in rabbit sera from vaccines after immunization with outer membrane vesicle ofNeisseria meningitidis serogroup B. Res. J. Microbiol., 3 (3): 105–113.

    Article  Google Scholar 

  • Costantino P., Norelli F., Giannozzi A., D’Ascenzi S., Bartoloni A., Kaur S.,et al. (1999). Size fractionation of bacterial capsular polysaccharides for their use in conjugate vaccines. Vaccine, 17 (9–10): 1251–1263.

    Article  CAS  PubMed  Google Scholar 

  • Claassen I., Meylis J., van der Ley P., Peeters C., Brons H., Robert J.,et al. (1996). Production, characterization and control of aNeisseria meningitidis hexavalent class 1 outer membrane protein containing vesicle vaccine. Vaccine., 14 (10): 1001–1008.

    Article  CAS  PubMed  Google Scholar 

  • Fleck R.A., Athwal H., Bygraves J.A., Hockley D.J., Feavers I.M., Stacey G.N. (2003). Optimization of nb-4 and hl-60 differentiation for use in opsonophagocytosis assays. In Vitro Cell Dev. Biol. Anim., 39 (5–6): 235–342.

    Article  PubMed  Google Scholar 

  • Jansen W.T., Vakevainen-Anttila M., Kayhty H., Nahm M., Bakker N., Verhoef J.,et al. (2001). Comparison of a classical phagocytosis assay and a flow cytometry assay for assessment of the phagocytic capacity of sera from adults vaccinated with a pneumococcal conjugate vaccine. Clin. Diagn. Lab. Immunol., 8 (2): 245–250.

    CAS  PubMed  Google Scholar 

  • Jin Z., Chu C., Robbins J.B., Schneerson R. (2003). Preparation and characterization of group A meningococcal capsular polysaccharide conjugates and evaluation of their immunogenicity in mice. Infect. Immun., 71: 5115–5120.

    Article  CAS  PubMed  Google Scholar 

  • Lehmann A.K., Sornes S., Halstensen A. (2000). Phagocytosis: measurement by flow cytometry. J. Immunol. Methods, 243 (1–2): 229–242.

    Article  CAS  PubMed  Google Scholar 

  • Li W., Chung S.C. (2003). Flow cytometric evaluation of leukocyte function in rat whole blood. In Vitro Cell Dev. Biol. Anim., 39 (10): 413–419.

    Article  PubMed  Google Scholar 

  • Lun A., Schmitt M., Renz H. (2000). Phagocytosis and oxidative burst: reference values for flow cytometric assays independent of age. Clin. Chem., 46 (11): 1836–1839.

    CAS  PubMed  Google Scholar 

  • Martinez J., Pilishvili T., Barnard S., Caba J., Spear W., Romero-Steiner S.,et al. (2002). Opsonophagocytosis of fluorescent polystyrene beads coupled toNeisseria meningitidis serogroup A, C, Y, or W135 polysaccharide correlates with serum bactericidal activity. Clin. Diagn. Lab. Immunol., 9 (2): 485–488.

    PubMed  Google Scholar 

  • Rezaei N., Aghamohammadi A., Siadat S.D., Nejati M., Hojat Ahmadi H., Moin M., Pourpak Z., Kamali S., Norouzian D., Tabaraei B., Read R.C. (2007). Serum bactericidal antibody response to serogroup C polysaccharide meningococcal vaccination in children with primary antibody deficiencies. Vaccine, 25: 5308–5314.

    Article  CAS  PubMed  Google Scholar 

  • Romero-Steiner S., Frasch C.E., Carlone G., Fleck R.A., Goldblatt D., Nahm M.H. (2006). Use of opsonophagocytosis for serological evaluation of pneumococcal vaccines. Clin. Vaccine Immunol., 13 (2): 165–169.

    Article  CAS  PubMed  Google Scholar 

  • Rosenstein N.E., Perkins B.A., Stephens D.S., Popovic T., Hughes J.M. (2001). Meningococcal disease. N. Engl. J. Med., 344 (18): 1378–1388.

    Article  CAS  PubMed  Google Scholar 

  • Schneerson R., Barrera O., Sutton A., Robbins J.B. (1980). Preparation, characterization, and immunogenicity ofHaemophilus influenzae type b polysaccharide-protein conjugates. J. Exp. Med., 152 (2): 361–376.

    Article  CAS  PubMed  Google Scholar 

  • Siadat S.D., Behzadiyannejad Q., Tabaraie B., Ahmadi H., Nejati M. (2006). Extraction and molecular evaluation ofNeisseria meningitidis serogroup B outer membrane vesicle containing PorA. Danshvar. Medicine, 65: 57–64.

    Google Scholar 

  • Siadat S.D., Norouzian D. (2007). Meningococcal vaccines: past, present, and future perspective. J. Infect. Develop. Country, 1 (2): 129–146.

    CAS  Google Scholar 

  • Siadat S.D., Behzadiannejad Q., Tabaraie B., Ahmadi H., Norouzian D., Najar-Peerayeh S., Nejati M., Hedayati M.H., Zangeneh M. (2007a). Evaluation of serum bactericidal activity specific forNeisseria meningitidis serogroup A and B: effect of immunization withNeisseria meningitidis serogroup A polysaccharide and serogroup B outer membrane vesicle conjugate as a bivalent meningococcus vaccine candidate. Res. J. Microbiol., 2 (5): 436–444.

    Article  Google Scholar 

  • Siadat S.D., Kheirandish M., Norouzian D., Behzadiyannejad Q., Najar Peerayeh S., Zangeneh M.,et al. (2007b). A flow cytometric opsonophagocytic assay for measurement of functional antibodies elicited after immunization with outer membrane vesicle ofNeisseria meningitidis serogroup B. Pakistan J. Biol. Sci., 10 (20): 3578–3584.

    Article  CAS  Google Scholar 

  • Siadat S.D., Norouzian D., Tabaraie B., Behzadiyannejad Q., Pirayeeh S.N., Nejati M.,et al. (2007c). Comparative studies of conjugated capsular polysaccharide ofNeisseria meningitidis serogroup A with outer membrane vesicle ofNeisseria meningitidis serogroup B. Res. J. Microbiol., 2 (4): 337–345.

    Article  CAS  Google Scholar 

  • Vakevainen M., Jansen W., Saeland E., Jonsdottir I., Snippe H., Verheul A.,et al. (2001). Are the opsonophagocytic activities of antibodies in infant sera measured by different pneumococcal phagocytosis assays comparable? Clin. Diagn. Lab. Immunol., 8 (2): 363–369.

    Google Scholar 

  • Valdivia R.H., Falkow S. (1998). Flow cytometry and bacterial pathogenesis. Curr. Opin. Microbiol., 1 (3):359–363.

    Article  CAS  PubMed  Google Scholar 

  • WHO — World Health Organization (1976). Requirements for meningococcal polysaccharide vaccine. Tech. Rep. Ser., 594: 50–75.

    Google Scholar 

Download references

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Authors and Affiliations

  1. Research Center of Iranian Blood Transfusion Organization, Tehran, Iran

    Maryam Kheirandish

  2. Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran

    Seyed Davar Siadat, Mohamad Reza Razavi, Mohammad Reza Aghasadeghi, Jalal Izadi Mobarakeh, Seyed Mehdi Sadat & Ali Sharifat Salmani

  3. Department of Bacterial Vaccines and Antigen Production, Pasteur Institute of Iran, Tehran, Iran

    Seyed Davar Siadat & Dariush Norouzian

  4. Growth and Development Research Center, Pediatrics Center of Excellence, Children’s Medical Center, Tehran University of Medical Sciences, Tehran, Iran

    Nima Rezaei

  5. Department of Cell and Molecular Biology, School of Biology, University of Tehran, Tehran, Iran

    Ali Farazmand

  6. Department of Infectious Diseases, Faculty of Medicine, Islamic Azad University, Tehran, Iran

    Mehrangiz Zangeneh

  7. Department of Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran

    Arfa Moshiri

Authors
  1. Maryam Kheirandish
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  2. Seyed Davar Siadat
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  3. Dariush Norouzian
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  4. Mohamad Reza Razavi
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  5. Mohammad Reza Aghasadeghi
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  6. Nima Rezaei
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  7. Ali Farazmand
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  8. Jalal Izadi Mobarakeh
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  9. Mehrangiz Zangeneh
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  10. Arfa Moshiri
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  11. Seyed Mehdi Sadat
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  12. Ali Sharifat Salmani
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Correspondence to Seyed Davar Siadat.

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Kheirandish, M., Siadat, S.D., Norouzian, D. et al. Measurement of opsonophagocytic activity of antibodies specific toNeisseria meningitidis serogroup A capsular polysaccharide-serogroup B outer membrane vesicle conjugate in animal model. Ann. Microbiol. 59, 801–806 (2009). https://doi.org/10.1007/BF03179226

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  • DOI: https://doi.org/10.1007/BF03179226

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Annals of Microbiology

ISSN: 1869-2044

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