Neisseria meningitidis serogroup C polysaccharide and serogroup B outer membrane vesicle conjugate as a bivalent meningococcus vaccine candidate

Fukasawa, Lucila Okuyama; Gorla, Maria Cecília Outeiro; Schenkman, Rocilda P.F.; Garcia, Ligiane R.; Carneiro, Sylvia Mendes; Raw, Isaı́as; Tanizaki, Martha M.

doi:10.1016/s0264-410x(99)00177-2

Cited by 19 publications

(20 citation statements)

References 16 publications

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“…The OMV must have the minimum amount of lipo-oligo-saccharide (LOS endotoxin), a constituent of these vesicles, in order to diminish the vaccine toxicity level, but maintaining vesicle integrity. In the Instituto Butantan the production process of N. meningitidis C (3,19); the evaluation of the importance of a second serogroup B strain as vaccine component (9); the obtainment of vesicles with appropriate characteristics (with IRP expression and with low level of LOS); the conjugation process of N. meningitidis C polysaccharide with N. meningitidis B OMV (12,13) were studied. In addition, it is worth mentioning that OMV also are employed as carriers of polysaccharides in conjugated vaccines against Haemophilus influenzae and in vaccines against pneumonia (2,11).…”

Section: Introductionmentioning

confidence: 99%

Production of outer membrane vesicles (OMV) in batch cultivation of Neisseria meningitidis serogroup B

Santos¹,

Baruque-Ramos

Tanizaki³

et al. 2006

Braz. J. Microbiol.

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Meningococcal disease is an important cause of death and morbidity throughout the world. Nearly 330,000 cases and 35,000 deaths occur yearly. Neisseria meningitidis, serogroup B strain N.44/89, is prevalent in Brazil. Its outer membrane vesicles (OMV) with iron regulated proteins (IRP) are released to the culture medium and are used as antigen for vaccine production. In order to have knowledge about the kinetic parameters, especially the final OMV concentration values, 20-h batch cultivations were carried out in Catlin medium with iron restriction. Process conditions comprised: 7 L bioreactor, 36ºC, 0.5 atm, overlay air flowrate of 1 L/min, agitation varying from 250 rpm to 850 rpm and dissolved oxygen control set at 10% of saturation condition. Biomass was determined by optical density at 540 nm and dry weight. Glycerol, lactate, pH and dissolved oxygen were measured from samples taken during cultivation. Outer membrane vesicle (OMV) concentration was determined by Lowry's method after ultracentrifugation. IRP presence was verified by SDS-PAGE. Highest biomass value, corresponding to the highest initial lactate concentration (7.84 g/L) was achieved at the 9 th hour process time corresponding to 1.0 g/L dry biomass and 2.3 optical density at 540 nm. Lactate consumption was directly related to cell growth (yield factor: 0.24 g dry biomass / g lactate). Glycerol concentration in the medium did not change significantly during the process. OMV concentration reached the highest value of 80 mg/L at end cultivation time. The obtained results suggest that lactate is a main limiting growth factor and the maximum amount of antigen is obtained during stationary growth and cell death phases.

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Section: Introductionmentioning

confidence: 99%

Production of outer membrane vesicles (OMV) in batch cultivation of Neisseria meningitidis serogroup B

Santos¹,

Baruque-Ramos

Tanizaki³

et al. 2006

Braz. J. Microbiol.

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“…PSC was conjugated with OMV from strain N44/89 and N603/95 using adipic acid dihydrazide (ADH) as a linker and 1‐ethyl‐3‐(3‐dimethylamino‐propyl)carbodiimide (EDAC) as coupling reagent as previously described [13]. The conjugate PSC was purified from free PSC by ultracentrifugation at 100,000 g for 3 h and OMV integrity after conjugation with PSC was checked by electronic microscopy [13]. The final conjugation yield was approximately 20% in PSC and the OMV protein/PS ratio (mg/mg) was about 4 for both conjugated OMV.…”

Section: Methodsmentioning

confidence: 99%

“…Antibodies rose against PSC and OMV were measured by the use of a previously described procedure [13]. For PSC the plates were pre‐coated with polylysine (5 μgml −1 in PBS, pH 7.2, for 30 min, at 37 °C) and washed with water.…”

Section: Methodsmentioning

confidence: 99%

Adjuvant can improve protection induced by OMV vaccine againstNeisseria meningitidisserogroups B/C in neonatal mice

Fukasawa

Dias

Schenkman

et al. 2004

FEMS Immunology &Amp; Medical Microbiology

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Meningococcal outer membrane vesicle (OMV) vaccines are weak antigens in infants. This study aimed at investigating alternative adjuvants for induction of functional antibodies in newborn mice. Serogroup B/C anti-meningococcal vaccines, consisting of capsular polysaccharide from serogroup C (PSC) conjugated to OMV from one serogroup B serosubtype prevalent in Brazil, combined with OMV from another prevalent serosubtype, were tested in newborn and adult mice with the following adjuvants: aluminum hydroxide, MPL (monophosphoryl lipid A), Titermax and MF59. Total IgG, IgG avidity index determination and bactericidal assay were performed with sera from immunized mice. Antibodies induced against PSC in newborn mice showed avidity and bactericidal titers, similar to those obtained in adult mice, independently of the adjuvant. Evidence is presented that the inclusion of MF59 enhanced the immune response against OMV in newborn mice.

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“…In a previous report, sera of mice immunized with serogroup C strain IMC 2135 were shown to react with a protein of approximately 190 kDa in OMV from the serogroup B strain N44/89, as determined by immunoblotting (Fukasawa et al, 1999). This protein was therefore first studied as a 190 kDa monomer (Fukasawa et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

“…However, it was shown that the recombinant protein obtained by Comanducci et al (2002) consists of 362 amino acids, including a possible leader peptide of 23 amino acids. The mature protein has a predicted molecular mass of 35 363 Da, and a leucine zipper structure probably maintains its pentameric conformation.In a previous report, sera of mice immunized with serogroup C strain IMC 2135 were shown to react with a protein of approximately 190 kDa in OMV from the serogroup B strain N44/89, as determined by immunoblotting (Fukasawa et al, 1999). This protein was therefore first studied as a 190 kDa monomer (Fukasawa et al, 2000).…”

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confidence: 99%

Immune response to native NadA from Neisseria meningitidis and its expression in clinical isolates in Brazil

Fukasawa

Gorla

Lemos

et al. 2003

Journal of Medical Microbiology

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A mAb against the NadA protein from Neisseria meningitidis strain 3006 (serosubtype B : 2b : P1.2 : P5.2,8) demonstrated strong bactericidal activity against Brazilian epidemic serogroup B strain N44/89 (B : 4,7 : P1.19,15 : P5.5,7) and a serogroup C strain, IMC 2135 (C : 2a : P1.5,2), but not against another serogroup C strain, N1002/90 (C : 2b : P1.3 : P5.8). The immunogenicity of native NadA in an outer-membrane vesicle (OMV) preparation was also tested. Serum from mice immunized with OMV from serogroup B strain N44/89, which contains the NadA protein, showed bactericidal activity against serogroup B and C strains possessing NadA. In dotblot analysis of 100 serogroup B and 100 serogroup C isolates from Brazilian patients, the mAb to NadA recognized about 60 % of the samples from both serogroups. The molecular mass of the NadA protein from strain N44/89 determined by mass spectrometry was 37 971 Da and the peptide sequences were identical to those of NadA from N. meningitidis strain MC58. INTRODUCTIONSuccessful vaccines against Neisseria meningitidis serogroups A, C, Y and W135 offer protection in adults and in children over 2 years of age (Frasch, 1995). However, some recent studies have shown immunological hyporesponsiveness to a second vaccination (Granoff et al., 1998; Richmond et al., 2000). Improved vaccines composed of capsular polysaccharide-protein conjugate antigens have been investigated and these vaccines have been shown to be protective for younger children (Zollinger, 1997). Clinical trials have been performed with capsular polysaccharides from meningococcal serogroups A and C conjugated with the CRM197, a mutant diphtheria toxin (Anderson et al., 1994;Twumasi et al., 1995). Extension of this approach to a group B meningococcal vaccine is not straightforward, since the group B capsular polysaccharide is a poor immunogen in humans (Poolman, 1995). Development of vaccines against serogroup B has therefore focused mostly on the use of outermembrane vesicles (OMV) containing different proteins (Blake & Gotschlich, 1986). Induction of protective immunity by these OMV vaccines has been studied in several trials and case-control studies. Trials in Chile and Brazil showed poor protection in children less than 4 years of age (Boslego et al., 1995; Milagres et al., 1994). Although efficacy in the range of 50-80 % in children over 4 years old was recorded (Anderson et al., 1994), improvements are clearly needed. Another limitation of OMV vaccines is that the protein antigens that induce bactericidal antibodies show sequence and antigenic variability. The bactericidal antibodies induced by OMV vaccines were found to be directed mainly against two classes of proteins, PorA (Poolman, 1995) and Opa (Rosenqvist et al., 1993). Therefore, interest has increased in finding conserved, surface-exposed proteins in serogroup B meningococcus that are able to induce bactericidal antibodies. NspA (Martin et al., 1997), GNA 33 (Pizza et al., 2000 and NadA (Comanducci et al., 2002), have been described as proteins that sa...

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Neisseria meningitidis serogroup C polysaccharide and serogroup B outer membrane vesicle conjugate as a bivalent meningococcus vaccine candidate

Cited by 19 publications

References 16 publications

Production of outer membrane vesicles (OMV) in batch cultivation of Neisseria meningitidis serogroup B

Production of outer membrane vesicles (OMV) in batch cultivation of Neisseria meningitidis serogroup B

Adjuvant can improve protection induced by OMV vaccine againstNeisseria meningitidisserogroups B/C in neonatal mice

Immune response to native NadA from Neisseria meningitidis and its expression in clinical isolates in Brazil

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