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Recombinant protein truncation strategy for inducing bactericidal antibodies to the macrophage infectivity potentiator protein of Neisseria meningitidis and circumventing potential cross-reactivity with human FK506-binding proteins

Recombinant protein truncation strategy for inducing bactericidal antibodies to the macrophage infectivity potentiator protein of Neisseria meningitidis and circumventing potential cross-reactivity with human FK506-binding proteins
Recombinant protein truncation strategy for inducing bactericidal antibodies to the macrophage infectivity potentiator protein of Neisseria meningitidis and circumventing potential cross-reactivity with human FK506-binding proteins
A recombinant (r) Macrophage Infectivity Potentiator (MIP) protein of Neisseria meningitidis induces significant serum bactericidal antibody production in mice and is a candidate meningococcal vaccine antigen. However, bioinformatics analysis of MIP showed some amino acid sequence similarity with human FK506-binding proteins (FKBPs) in residues 166-252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognise human proteins, we immunised mice with recombinant Truncated Type I rMIP proteins that lacked the globular domain and the signal leader peptide (LP) signal sequence (amino acids 1-22) and contained the HIS purification tag at either the N- or C-terminus. The immunogenicity of Truncated rMIP proteins was compared to Full rMIP proteins (containing the globular domain) with either a N- or C-terminal HIS tag and with or without the LP sequence. By comparing the functional murine antibody responses to these various constructs, we identified that C-term HIS Truncated rMIP (-LP) delivered in liposomes induced high levels of antibodies that bound to the surface of wild-type but not ?mip mutant meningococci and showed bactericidal activity against homologous Type I MIP (median titres of 128-256) and heterologous Type II and III (median titres of 256-512) strains, thereby providing at least 82% serogroup B strain coverage. In contrast, in constructs lacking the LP, placement of the HIS tag at the N-terminus appeared to abrogate bactericidal activity. The strategy used in this study would obviate any potential concerns regarding the use of MIP antigens for inclusion in bacterial vaccines
0019-9567
730-742
Bielecka, Magdalena K.
90391ea3-aa1f-4104-a893-568c138718a2
Devos, Nathalie
376b1648-5ac2-4c5e-92c7-9cf228fda87c
Gilbert, Melanie
1b4ccf7d-283d-4122-ac86-54f2eaac9321
Hung, Miao-Chiu
53d4bf2c-4e0e-4c77-9385-218350560fdb
Weynants, Vincent
36b1f098-a1fb-444f-acb1-d85009cc6d4c
Heckels, John E.
fcfcfafe-5ca8-4728-9c5e-cb67f9af7e31
Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078
Bielecka, Magdalena K.
90391ea3-aa1f-4104-a893-568c138718a2
Devos, Nathalie
376b1648-5ac2-4c5e-92c7-9cf228fda87c
Gilbert, Melanie
1b4ccf7d-283d-4122-ac86-54f2eaac9321
Hung, Miao-Chiu
53d4bf2c-4e0e-4c77-9385-218350560fdb
Weynants, Vincent
36b1f098-a1fb-444f-acb1-d85009cc6d4c
Heckels, John E.
fcfcfafe-5ca8-4728-9c5e-cb67f9af7e31
Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078

Bielecka, Magdalena K., Devos, Nathalie, Gilbert, Melanie, Hung, Miao-Chiu, Weynants, Vincent, Heckels, John E. and Christodoulides, Myron (2015) Recombinant protein truncation strategy for inducing bactericidal antibodies to the macrophage infectivity potentiator protein of Neisseria meningitidis and circumventing potential cross-reactivity with human FK506-binding proteins. Infection and Immunity, 83 (2), 730-742. (doi:10.1128/IAI.01815-14). (PMID:25452551)

Record type: Article

Abstract

A recombinant (r) Macrophage Infectivity Potentiator (MIP) protein of Neisseria meningitidis induces significant serum bactericidal antibody production in mice and is a candidate meningococcal vaccine antigen. However, bioinformatics analysis of MIP showed some amino acid sequence similarity with human FK506-binding proteins (FKBPs) in residues 166-252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognise human proteins, we immunised mice with recombinant Truncated Type I rMIP proteins that lacked the globular domain and the signal leader peptide (LP) signal sequence (amino acids 1-22) and contained the HIS purification tag at either the N- or C-terminus. The immunogenicity of Truncated rMIP proteins was compared to Full rMIP proteins (containing the globular domain) with either a N- or C-terminal HIS tag and with or without the LP sequence. By comparing the functional murine antibody responses to these various constructs, we identified that C-term HIS Truncated rMIP (-LP) delivered in liposomes induced high levels of antibodies that bound to the surface of wild-type but not ?mip mutant meningococci and showed bactericidal activity against homologous Type I MIP (median titres of 128-256) and heterologous Type II and III (median titres of 256-512) strains, thereby providing at least 82% serogroup B strain coverage. In contrast, in constructs lacking the LP, placement of the HIS tag at the N-terminus appeared to abrogate bactericidal activity. The strategy used in this study would obviate any potential concerns regarding the use of MIP antigens for inclusion in bacterial vaccines

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More information

Accepted/In Press date: 24 November 2014
e-pub ahead of print date: 1 December 2014
Published date: February 2015
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 373406
URI: https://eprints.soton.ac.uk/id/eprint/373406
ISSN: 0019-9567
PURE UUID: cc83d413-fb32-4ccf-9329-c95fa4e56fea
ORCID for Miao-Chiu Hung: ORCID iD orcid.org/0000-0001-5877-8075

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Date deposited: 16 Jan 2015 13:15
Last modified: 06 Jun 2018 12:21

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Contributors

Author: Nathalie Devos
Author: Melanie Gilbert
Author: Miao-Chiu Hung ORCID iD
Author: Vincent Weynants
Author: John E. Heckels

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