The Dual Role of Lipids of the Lipoproteins in Trumenba, a Self-Adjuvanting Vaccine Against Meningococcal Meningitis B Disease.

Yin Luo,Gary Zlotnick,Sandeep Kumar,Emilia Byrne,Michael Schlittler,Annaliesa S Anderson,Ann Aulabaugh,Eugene Vidunas,Stephen W Raso,Elena Novikova,Jason C Rouse,Lakshmi Khandke,Kathrin U Jansen,Donald Stano,Thomas Gore,Robert L Dufield,Olga V Friese,Herbert A Runnels

doi:10.1208/s12248-016-9979-x

Yin Luo, Gary Zlotnick + Show 16 more

Open Access

https://doi.org/10.1208/s12248-016-9979-x

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Journal: AAPS PharmSci	Publication Date: Sep 7, 2016
Citations: 57	License type: CC BY 4.0

Affiliation: Pfizer (United States)

Abstract

Trumenba (bivalent rLP2086) is a vaccine licensed for the prevention of meningococcal meningitis disease caused by Neisseria meningitidis serogroup B (NmB) in individuals 10-25years of age in the USA. The vaccine is composed of two factor H binding protein (fHbp) variants that were recombinantly expressed in Escherichia coli as native lipoproteins: rLP2086-A05 and rLP2086-B01. The vaccine was shown to induce potent bactericidal antibodies against a broad range of NmB isolates expressing fHbp that were different in sequence from the fHbp vaccine antigens. Here, we describe the characterization of the vaccine antigens including the elucidation of their structure which is characterized by two distinct motifs, the polypeptide domain and the N-terminal lipid moiety. In the vaccine formulation, the lipoproteins self-associate to form micelles driven by the hydrophobicity of the lipids and limited by the size of the folded polypeptides. The micelles help to increase the structural stability of the lipoproteins in the absence of bacterial cell walls. Analysis of the lipoproteins in Toll-like receptor (TLR) activation assays revealed their TLR2 agonist activity. This activity was lost with removal of the O-linked fatty acids, similar to removal of all lipids, demonstrating that this moiety plays an adjuvant role in immune activation. The thorough understanding of the structure and function of each moiety of the lipoproteins, as well as their relationship, lays the foundation for identifying critical parameters to guide vaccine development and manufacture.

Highlights

Neisseria meningitidis, called meningococcus (Men), is a human commensal Gram-negative bacterium that can cause severe disease with substantial mortality and morbidity
The results described above support the perspective that the presence of the O-linked fatty acids in the tri-acylated N. meningitidis serogroup B (NmB) rLP2086 lipoproteins is critical for the stimulation of Toll-like receptor 2 (TLR2)/TLR1-expressing cells and, explain the immune enhancement observed with the lipidated forms of factor H binding protein (fHbp) compared to their non-lipidated forms
Trumenba is a well-characterized vaccine composed of two recombinant bacterial lipoproteins, NmB rLP2086-A05 and rLP2086-B01, which are tri-acylated with fatty acids of 14–19 carbon atoms in length

Summary

Introduction

Called meningococcus (Men), is a human commensal Gram-negative bacterium that can cause severe disease with substantial mortality and morbidity. N. meningitidis is categorized by its capsular polysaccharides into 12 serogroups, of which six cause the majority of disease [1]. Four of the serogroups (A, C, Y, W) can be controlled with capsular polysaccharidebased vaccines. A capsular polysaccharide-based vaccine approach was not feasible for protection against N. meningitidis serogroup B (NmB) [2] due to the low immunogenicity of the serogroup B capsular polysaccharide [3], which is similar to structures found on human neuronal cells [4]. NmB causes outbreaks in various settings and regions. To control these outbreaks, vaccines based on the highly variable porin A protein were used, but

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