Neisseria meningitidis causes explosive epidemics in sub-Saharan Africa. Most are caused by capsular group A strains. However, group W-135 and X strains also cause epidemics in this region, and these strains may emerge once mass immunization with a group A polysaccharide-protein conjugate vaccine is introduced. Our goal is to develop a meningococcal vaccine for Africa that targets strains from all capsular groups. Our approach will be to use novel protein antigens identified for "group B vaccines", which also elicit protective antibodies against strains with other capsules. These "unconventional" antigens will be presented in simple outer membrane vesicles (OMV) that have potent natural adjuvants. Our studies will build on previous experience with detergent-extracted OMV vaccines, which are proven to be safe and effective in humans. Their major limitation is that they elicit serum bactericidal antibodies primarily directed at PorA, which is antigenically variable. To extend protection to strains with heterologous PorA, we prepared mutants of group B strains that were engineered to over-express factor H binding protein (fHbp), which is a novel antigen in two promising group B recombinant protein vaccines. By introducing an additional mutation in LPS biosynthesis, we attenuated endotoxin activity. In mice, non-detergent-treated OMV vaccines prepared from the mutants elicited serum bactericidal antibody responses against genetically diverse group B strains, as well as epidemic group A, W-135 and X strains from Africa. Our hypothesis is that a native OMV vaccine prepared from mutant strains from Africa will elicit even broader bactericidal antibodies directed at PorA, fHbp and other antigens expressed by strains from Africa. Further, the LPS mutation will eliminate the need for detergent extraction of the OMV, which is used to decrease LPS in conventional OMV vaccines, but also extracts desirable antigens such as fHbp.
In Aim 1, we will investigate genetic lineages and sequence diversity of genes encoding fHbp, PorA and other vaccine antigens among 200 meningococcal isolates from a geographically diverse collection of strains from Africa.
In Aim 2, we will measure antigen expression by a quantitative capture ELISA, and antigen surface-accessibility on live bacteria by flow cytometry.
In Aim 3, we will create mutants of recent African epidemic strains, which will be engineered to express more than one PorA molecule, over-express fHbp, and have attenuated endotoxin. The vaccine strains also will be selected for naturally high expression of an adhesin/invasin, NadA. We will prepare native OMV vaccines from the mutants, and assess OMV toxicity by measuring inflammatory cytokine responses of human PBMCs incubated in vitro with the vaccine. We will immunize mice and infant primates and measure serum bactericidal antibody responses against strains from Africa. The results will provide proof of principle that the OMV vaccine is likely to be well-tolerated in humans and elicit broad protective immunity. These findings would support an application to test the OMV vaccine in humans for control of meningococcal epidemics in sub-Sahara caused by strains from all capsular groups.

Public Health Relevance

Meningococci cause explosive epidemics of meningitis in sub-Sahara Africa that can involve more than 100,000 cases in a few months. Most epidemics have been caused by encapsulated group A strains but strains from other capsular groups also have begun to cause epidemics in the region. A polysaccharide conjugate vaccine against group A disease is being developed for Africa but there is grave concern that strains with other capsules may emerge and cause epidemics once mass immunization with the group A conjugate vaccine is introduced. We propose to develop an outer membrane vesicle vaccine from mutant meningococcal strains, engineered for over-expression of promising protein vaccine antigens, as a universal meningococcal vaccine for Africa against disease caused by strains from all capsular groups.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI082263-03
Application #
8204737
Study Section
Vaccines Against Microbial Diseases (VMD)
Program Officer
Taylor, Christopher E,
Project Start
2010-01-01
Project End
2013-12-31
Budget Start
2012-01-01
Budget End
2012-12-31
Support Year
3
Fiscal Year
2012
Total Cost
$650,640
Indirect Cost
$187,913
Name
Children's Hospital & Res Ctr at Oakland
Department
Type
DUNS #
076536184
City
Oakland
State
CA
Country
United States
Zip Code
94609
Beernink, Peter T; Shaughnessy, Jutamas; Stefek, Heather et al. (2014) Heterogeneity in rhesus macaque complement factor H binding to meningococcal factor H binding protein (FHbp) informs selection of primates to assess immunogenicity of FHbp-based vaccines. Clin Vaccine Immunol 21:1505-11
Shaughnessy, Jutamas; Vu, David M; Punjabi, Rahi et al. (2014) Fusion protein comprising factor H domains 6 and 7 and human IgG1 Fc as an antibacterial immunotherapeutic. Clin Vaccine Immunol 21:1452-9
Lewis, Lisa A; Vu, David M; Granoff, Dan M et al. (2014) Inhibition of the alternative pathway of nonhuman infant complement by porin B2 contributes to virulence of Neisseria meningitidis in the infant rat model. Infect Immun 82:2574-84
Costa, Isabella; Pajon, Rolando; Granoff, Dan M (2014) Human factor H (FH) impairs protective meningococcal anti-FHbp antibody responses and the antibodies enhance FH binding. MBio 5:e01625-14
Lewis, Lisa A; Vu, David M; Vasudhev, Shreekant et al. (2013) Factor H-dependent alternative pathway inhibition mediated by porin B contributes to virulence of Neisseria meningitidis. MBio 4:e00339-13
Giuntini, Serena; Vu, David M; Granoff, Dan M (2013) fH-dependent complement evasion by disease-causing meningococcal strains with absent fHbp genes or frameshift mutations. Vaccine 31:4192-9
Rossi, Raffaella; Granoff, Dan M; Beernink, Peter T (2013) Meningococcal factor H-binding protein vaccines with decreased binding to human complement factor H have enhanced immunogenicity in human factor H transgenic mice. Vaccine 31:5451-7
Granoff, Dan M; Ram, Sanjay; Beernink, Peter T (2013) Does binding of complement factor H to the meningococcal vaccine antigen, factor H binding protein, decrease protective serum antibody responses? Clin Vaccine Immunol 20:1099-107
Konar, Monica; Granoff, Dan M; Beernink, Peter T (2013) Importance of inhibition of binding of complement factor H for serum bactericidal antibody responses to meningococcal factor H-binding protein vaccines. J Infect Dis 208:627-36
Giuntini, Serena; Beernink, Peter T; Reason, Donald C et al. (2012) Monoclonal antibodies to meningococcal factor H binding protein with overlapping epitopes and discordant functional activity. PLoS One 7:e34272

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