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Infection and Immunity, June 2001, p. 3809-3816, Vol. 69, No. 6
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.6.3809-3816.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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

Keith A. Jolley,dagger Lynn Appleby, J. Claire Wright,Dagger Myron Christodoulides, and John E. Heckels*

Molecular Microbiology Group, Division of Cell and Molecular Medicine, University of Southampton Medical School, Southampton General Hospital, Southampton SO16 6YD, United Kingdom

Received 2 November 2000/Returned for modification 8 January 2001/Accepted 14 March 2001

The opc gene from Neisseria meningitidis was cloned into the pRSETA vector, and recombinant protein was expressed at high levels in Escherichia coli. The protein was readily purified by affinity chromatography and used for immunization with conventional Al(OH)3 adjuvant or after incorporation into liposomes and Zwittergent micelles. The resulting sera were analyzed for their ability to recognize purified recombinant protein and "native" protein in an enzyme immunoassay with outer membranes and by whole-cell immunofluorescence. Immunization with Al(OH)3 induced high levels of antibodies which reacted with the purified protein but did not recognize whole cells. In contrast, liposomes and micelles induced antibodies which reacted with the native protein in whole cells. The addition of monophosphoryl lipid A (MPLA) to either liposomes or micelle preparations increased the magnitude of the immune response and induced a wider range of immunoglobulin subclasses. This was associated with the ability of the sera to induce complement-mediated killing of the homologous strain. The most effective bactericidal activity was observed with Opc protein incorporated into liposomes containing MPLA. The magnitude of the bactericidal effect was strongly influenced by the level of expression of the Opc protein and was abolished by limited variation in the sequence of the protein expressed by heterologous strains.

* Corresponding author. Mailing address: Molecular Microbiology Group, Division of Cell and Molecular Medicine, Mailpoint 814, Southampton General Hospital, Tremona Road, Southampton SO16 6YD, United Kingdom. Phone: 44-023-80796974. Fax: 44-023-80796992. E-mail: jeh{at}soton.ac.uk.

dagger Present address: Wellcome Trust Centre for the Epidemiology of Infectious Disease, Department of Zoology, University of Oxford, Oxford OX1 3PS, United Kingdom.

Dagger Present address: Department of Paediatrics, University of Oxford, John Radcliffe Hospital, Headington, Oxford OX3 9DU, United Kingdom.

Infection and Immunity, June 2001, p. 3809-3816, Vol. 69, No. 6
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.6.3809-3816.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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