Liposomes or traditional adjuvants: induction of bactericidal activity by the macrophage infectivity potentiator protein (Mip) ofNeisseria meningitidis

Apmis ◽  
2017 ◽  
Vol 125 (8) ◽  
pp. 725-731 ◽  
Author(s):  
Liliana Costoya ◽  
Juan Marzoa ◽  
Carlos Ferreirós ◽  
Maria Teresa Criado
Keyword(s):  
Bactericidal Activity ◽  
Macrophage Infectivity Potentiator ◽  
Macrophage Infectivity

scholarly journals 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

2014 ◽  
Vol 83 (2) ◽  
pp. 730-742 ◽  
Author(s):  
Magdalena K. Bielecka ◽  
Nathalie Devos ◽  
Mélanie Gilbert ◽  
Miao-Chiu Hung ◽  
Vincent Weynants ◽  
...  
Keyword(s):  
Neisseria Meningitidis ◽  
Bactericidal Activity ◽  
Binding Proteins ◽  
Leader Peptide ◽  
Type I ◽  
Globular Domain ◽  
His Tag ◽  
Macrophage Infectivity Potentiator ◽  
Fk506 Binding Proteins ◽  
Macrophage Infectivity

A recombinant macrophage infectivity potentiator (rMIP) protein ofNeisseria meningitidisinduces 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 to human FK506-binding proteins (FKBPs) in residues 166 to 252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognize human proteins, we immunized mice with recombinant truncated type I rMIP proteins that lacked the globular domain and the signal leader peptide (LP) signal sequence (amino acids 1 to 22) and contained the His purification tag at either the N or C terminus (C-term). The immunogenicity of truncated rMIP proteins was compared to that of full (i.e., full-length) rMIP proteins (containing the globular domain) with either an 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 determined 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 Δmipmutant meningococci and showed bactericidal activity against homologous type I MIP (median titers of 128 to 256) and heterologous type II and III (median titers of 256 to 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.

scholarly journals The Neisseria meningitidis Macrophage Infectivity Potentiator Protein Induces Cross-Strain Serum Bactericidal Activity and Is a Potential Serogroup B Vaccine Candidate

2011 ◽  
Vol 79 (9) ◽  
pp. 3784-3791 ◽  
Author(s):  
Miao-Chiu Hung ◽  
Omar Salim ◽  
Jeannette N. Williams ◽  
John E. Heckels ◽  
Myron Christodoulides
Keyword(s):  
Neisseria Meningitidis ◽  
Bactericidal Activity ◽  
Legionella Pneumophila ◽  
Type I ◽  
Content Type ◽  
Serum Bactericidal Activity ◽  
Monophosphoryl Lipid A ◽  
Macrophage Infectivity Potentiator ◽  
Sequence Types ◽  
Macrophage Infectivity

ABSTRACTA gene encoding a 29-kDa protein fromNeisseria meningitidisserogroup B strain MC58 with homology to the macrophage infectivity potentiator (MIP) protein ofLegionella pneumophilawas cloned and expressed inEscherichia coli, and the purified soluble recombinant protein (rMIP) was used for immunization studies. Analysis of the predicted amino acid sequences of MIP from 13 well-characterized meningococcal strains, isolated from carriers or patients and differing in serogroup, serotype, and subtype, showed that the protein was highly conserved (98 to 100%), with only three distinct sequence types (designated I, II, and III) found. Western blotting showed that the MIP protein was expressed at similar levels by all of these strains. Immunization of mice with type I MC58 rMIP in detergent micelles and liposomes containing monophosphoryl lipid A (MPLA) induced high levels of surface-reactive antibodies with serum bactericidal activity (SBA) titers of 1/1,024 against the homologous strain. Bactericidal antibodies were also induced with the protein in saline alone and liposomes alone (titers, 1/128) but not following adsorption to Al(OH)3. Significantly, antisera raised against type I rMIP administered in saline or liposomes killed strains of heterologous sequence types II and III with similar SBA titers (1/128 to 1/256). Taken together, these findings suggest that rMIP can provide cross-strain protection against meningococci and should be considered a potential antigen for inclusion in new vaccines against meningococcal infection.

Inhibitors of macrophage infectivity potentiator-like PPIases affect neisserial and chlamydial pathogenicity

2016 ◽  
Vol 48 (4) ◽  
pp. 401-408 ◽  
Author(s):  
Anastasija Reimer ◽  
Florian Seufert ◽  
Matthias Weiwad ◽  
Jutta Ebert ◽  
Nicole M. Bzdyl ◽  
...  
Keyword(s):  
Macrophage Infectivity Potentiator ◽  
Macrophage Infectivity

Pharmacophore based virtual screening for identification of marine bioactive compounds as inhibitors against macrophage infectivity potentiator (Mip) protein of Chlamydia trachomatis

RSC Advances ◽  
2016 ◽  
Vol 6 (23) ◽  
pp. 18946-18957 ◽  
Author(s):  
Vijayan Ramachandran ◽  
Elavarasi Padmanaban ◽  
Kalaiarasan Ponnusamy ◽  
Subbarao Naidu ◽  
Manoharan Natesan
Keyword(s):  
Chlamydia Trachomatis ◽  
Virtual Screening ◽  
Sexually Transmitted Diseases ◽  
Bioactive Compounds ◽  
Virulence Factor ◽  
Sexually Transmitted ◽  
Macrophage Infectivity Potentiator ◽  
Macrophage Infectivity

Macrophage infectivity potentiator (Mip) is the virulence factor fromChlamydia trachomatisthat is primarily responsible for causing sexually transmitted diseases (STDs) and blindness.

scholarly journals Synthesis in Escherichia coli of two smaller enzymically active analogues of Coxiella burnetii macrophage infectivity potentiator (CbMip) protein utilizing a single open reading frame from the cbmip gene

1998 ◽  
Vol 335 (1) ◽  
pp. 67-77 ◽  
Author(s):  
Yin-yuan MO ◽  
J. SESHU ◽  
Dong WANG ◽  
Louis P. MALLAVIA
Keyword(s):  
Escherichia Coli ◽  
Coxiella Burnetii ◽  
Sequence Homology ◽  
Degradation Products ◽  
Open Reading Frame ◽  
Site Directed Mutagenesis ◽  
Ppiase Activity ◽  
Reading Frame ◽  
Macrophage Infectivity Potentiator ◽  
Macrophage Infectivity

FK506-binding proteins (FKBPs) have been identified in a variety of eukaryotic and prokaryotic organisms. Macrophage infectivity potentiator (CbMip, 23.5 kDa) protein of the obligate intracellular bacterium, Coxiella burnetii, was shown previously to belong to the family of FKBPs based on sequence homology and peptidyl-prolyl cis/trans isomerase (PPIase) activity. Further characterization of the cbmip gene has identified two additional proteins with molecular masses of 15.5 and 15.0 kDa that are synthesized, in addition to the 23.5 kDa CbMip, when expressed in Escherichia coli. Amino acid sequencing at the N-terminus combined with transcription and translation fusion expression revealed that the two proteins were synthesized from the same open reading frame of the cbmip gene, but starting at different internal translation start codons, probably by translational reinitiation. When the internal methionines serving as start sites were replaced with lysine by site-directed mutagenesis, the synthesis of 15.5 and 15.0 kDa proteins was abolished even though the synthesis of 23.5 kDa CbMip was intact. This confirmed that the 15.5 and 15.0 kDa proteins are indeed generated by translational reinitiation and are not degradation products of the 23.5 kDa protein. Like other FKBPs, both 15.5 and 15.0 kDa proteins exhibit PPIase activity. Because they share significant sequence homology with FKBPs and have a similar PPIase activity, 15.5 and 15.0 kDa proteins are designated as C. burnetiiFKBP (Cb-FKBP) analogues I and II, respectively. TnphoA mutagenesis demonstrated that whereas the large protein (CbMip) is secreted, Cb-FKBP analogues I and II are cytoplasmic, indicating that structural variations could allow for different subcellular compartmentalization of similar proteins. Western-blot analysis of lysates of purified C. burnetii using a CbMip-specific monoclonal antibody revealed the presence of a protein migrating at ≈ 15 kDa, indicating the presence of smaller Cb-FKBP analogue(s) in C. burnetii, although at much lower levels compared with 23.5 kDa CbMip. This unique gene organization seen with cbmip may provide the organism with a mechanism of efficient use of its limited genetic information to synthesize proteins that are structurally different yet functionally similar.

scholarly journals Legionella dresdenensis sp. nov., isolated from river water

2010 ◽  
Vol 60 (11) ◽  
pp. 2557-2562 ◽  
Author(s):  
Paul Christian Lück ◽  
Enno Jacobs ◽  
Isolde Röske ◽  
Ute Schröter-Bobsin ◽  
Roger Dumke ◽  
...  
Keyword(s):  
Type Strain ◽  
Biochemical Characterization ◽  
Novel Species ◽  
Phenotypic Characterization ◽  
Fatty Acid Profiles ◽  
Sequence Comparisons ◽  
River Elbe ◽  
Macrophage Infectivity Potentiator ◽  
Macrophage Infectivity

Legionella-like isolates, strains W03-356T, W03-357 and W03-359, from three independent water samples from the river Elbe, Germany, were analysed by using a polyphasic approach. Morphological and biochemical characterization revealed that they were Gram-negative, aerobic, non-spore-forming bacilli with a cut glass colony appearance that grew only on l-cysteine-supplemented buffered charcoal yeast extract agar. Phylogenetic analysis based on sequence comparisons of the 16S rRNA, macrophage infectivity potentiator (mip), gyrase subunit A (gyrA), ribosomal polymerase B (rpoB) and RNase P (rnpB) genes confirmed that the three isolates were distinct from recognized species of the genus Legionella. Phenotypic characterization of strain W03-356T based on fatty acid profiles confirmed that it was closely related to Legionella rubrilucens ATCC 35304T and Legionella pneumophila ATCC 33152T, but distinct from other species of the genus Legionella. Serotyping of the isolates showed that they were distinct from all recognized species of the genus Legionella. Strains W03-356T, W03-357 and W03-359 are thus considered to represent a novel species of the genus Legionella, for which the name Legionella dresdenensis sp. nov. is proposed. The type strain is W03-356T (=DSM 19488T=NCTC 13409T).

scholarly journals Molecular Characterization and Subcellular Localization of Macrophage Infectivity Potentiator, a Chlamydia trachomatis Lipoprotein

2007 ◽  
Vol 189 (13) ◽  
pp. 4739-4748 ◽  
Author(s):  
Laurence Neff ◽  
Sawsan Daher ◽  
Patrick Muzzin ◽  
Ursula Spenato ◽  
Fazil Gülaçar ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Chlamydia Trachomatis ◽  
Subcellular Localization ◽  
Palmitic Acid ◽  
Mass Spectrometry Analysis ◽  
Signal Peptidase ◽  
Gas Chromatography Mass Spectrometry ◽  
Posttranslational Processing ◽  
Macrophage Infectivity Potentiator ◽  
Macrophage Infectivity

ABSTRACT Macrophage infectivity potentiator (MIP) was originally reported to be a chlamydial lipoprotein from experiments showing incorporation of radiolabeled palmitic acid into native and recombinant MIP; inhibition of posttranslational processing of recombinant MIP by globomycin, known to inhibit signal peptidase II; and solubility of native MIP in Triton X-114. However, the detailed structural characterization of the lipid moiety on MIP has never been fully elucidated. In this study, bioinformatics and mass spectrometry analysis, as well as radiolabeling and immunochemical experiments, were conducted to further characterize MIP structure and subcellular localization. In silico analysis showed that the amino acid sequence of MIP is conserved across chlamydial species. A potential signal sequence with a contained lipobox was identified, and a recombinant C20A variant was prepared by replacing the probable lipobox cysteine with an alanine. Both incorporation of U-14C-esterified glycerol and [U-14C]palmitic acid and posttranslational processing that was inhibitable by globomycin were observed for recombinant wild-type MIP but not for the recombinant C20A MIP variant. The fatty acid contents of native and recombinant MIP were analyzed by gas chromatography-mass spectrometry, and the presence of amide-linked fatty acids in recombinant MIP was investigated by alkaline methanolysis. These results demonstrated a lipid modification in MIP similar to that of other prokaryotic lipoproteins. In addition, MIP was detected in an outer membrane preparation of Chlamydia trachomatis elementary bodies and was shown to be present at the surfaces of elementary bodies by surface biotinylation and surface immunoprecipitation experiments.

Sign in / Sign up

Export Citation Format

Share Document