The Lassa virus glycoprotein precursor GP-C is proteolytically processed by subtilase SKI-1/S1P

ABSTRACT Recovery from Lassa virus (LASV) infection usually precedes the appearance of neutralizing antibodies, indicating that cellular immunity plays a primary role in viral clearance. To date, the role of LASV-specific CD8+ T cells has not been evaluated in humans. To facilitate such studies, we utilized a predictive algorithm to identify candidate HLA-A2 supertype epitopes from the LASV nucleoprotein and glycoprotein precursor (GPC) genes. We identified three peptides (GPC42-50, GLVGLVTFL; GPC60-68, SLYKGVYEL; and GPC441-449, YLISIFLHL) that displayed high-affinity binding (≤98 nM) to HLA-A*0201, induced CD8+ T-cell responses of high functional avidity in HLA-A*0201 transgenic mice, and were naturally processed from native LASV GPC in human HLA-A*0201-positive target cells. HLA-A*0201 mice immunized with either GPC42-50 or GPC60-68 were protected against challenge with a recombinant vaccinia virus that expressed LASV GPC. The epitopes identified in this study represent potential diagnostic reagents and candidates for inclusion in epitope-based vaccine constructs. Our approach is applicable to any pathogen with existing sequence data, does not require manipulation of the actual pathogen or access to immune human donors, and should therefore be generally applicable to category A through C agents and other emerging pathogens.

Download Full-text

Signal peptide of Lassa virus glycoprotein GP-C exhibits an unusual length

FEBS Letters ◽

10.1016/s0014-5793(03)00160-1 ◽

2003 ◽

Vol 538 (1-3) ◽

pp. 203-206 ◽

Cited By ~ 76

Author(s):

Robert Eichler ◽

Oliver Lenz ◽

Thomas Strecker ◽

Wolfgang Garten

Keyword(s):

Signal Peptide ◽

Lassa Virus ◽

Virus Glycoprotein

Download Full-text

A Single Dose of Modified Vaccinia Ankara Expressing Lassa Virus-like Particles Protects Mice from Lethal Intra-cerebral Virus Challenge

Pathogens ◽

10.3390/pathogens8030133 ◽

2019 ◽

Vol 8 (3) ◽

pp. 133 ◽

Cited By ~ 8

Author(s):

Maria S. Salvato ◽

Arban Domi ◽

Camila Guzmán-Cardozo ◽

Sandra Medina-Moreno ◽

Juan Carlos Zapata ◽

...

Keyword(s):

Single Dose ◽

Matrix Protein ◽

Lethal Dose ◽

Health Impact ◽

Lassa Virus ◽

Protein Z ◽

Glycoprotein Precursor ◽

Lethal Challenge ◽

Virus Like Particles ◽

Virus Challenge

Lassa fever surpasses Ebola, Marburg, and all other hemorrhagic fevers except Dengue in its public health impact. Caused by Lassa virus (LASV), the disease is a scourge on populations in endemic areas of West Africa, where reported incidence is higher. Here, we report construction, characterization, and preclinical efficacy of a novel recombinant vaccine candidate GEO-LM01. Constructed in the Modified Vaccinia Ankara (MVA) vector, GEO-LM01 expresses the glycoprotein precursor (GPC) and zinc-binding matrix protein (Z) from the prototype Josiah strain lineage IV. When expressed together, GP and Z form Virus-Like Particles (VLPs) in cell culture. Immunogenicity and efficacy of GEO-LM01 was tested in a mouse challenge model. A single intramuscular dose of GEO-LM01 protected 100% of CBA/J mice challenged with a lethal dose of ML29, a Mopeia/Lassa reassortant virus, delivered directly into the brain. In contrast, all control animals died within one week. The vaccine induced low levels of antibodies but Lassa-specific CD4+ and CD8+ T cell responses. This is the first report showing that a single dose of a replication-deficient MVA vector can confer full protection against a lethal challenge with ML29 virus.

Download Full-text

Lassa Virus Glycoprotein Signal Peptide Displays a Novel Topology with an Extended Endoplasmic Reticulum Luminal Region

Journal of Biological Chemistry ◽

10.1074/jbc.m312975200 ◽

2004 ◽

Vol 279 (13) ◽

pp. 12293-12299 ◽

Cited By ~ 24

Author(s):

Robert Eichler ◽

Oliver Lenz ◽

Thomas Strecker ◽

Markus Eickmann ◽

Hans-Dieter Klenk ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Signal Peptide ◽

Lassa Virus ◽

Virus Glycoprotein ◽

Luminal Region

Download Full-text

Enhanced Efficacy of a Codon-Optimized DNA Vaccine Encoding the Glycoprotein Precursor Gene of Lassa Virus in a Guinea Pig Disease Model When Delivered by Dermal Electroporation

Vaccines ◽

10.3390/vaccines1030262 ◽

2013 ◽

Vol 1 (3) ◽

pp. 262-277 ◽

Cited By ~ 21

Author(s):

Kathleen Cashman ◽

Kate Broderick ◽

Eric Wilkinson ◽

Carl Shaia ◽

Todd Bell ◽

...

Keyword(s):

Guinea Pig ◽

Dna Vaccine ◽

Disease Model ◽

Lassa Virus ◽

Glycoprotein Precursor

Download Full-text

Amino acids from both N-terminal hydrophobic regions of the Lassa virus envelope glycoprotein GP-2 are critical for pH-dependent membrane fusion and infectivity

Journal of General Virology ◽

10.1099/vir.0.82950-0 ◽

2007 ◽

Vol 88 (8) ◽

pp. 2320-2328 ◽

Cited By ~ 42

Author(s):

Christian Klewitz ◽

Hans-Dieter Klenk ◽

Jan ter Meulen

Keyword(s):

Amino Acids ◽

Structural Changes ◽

Point Mutations ◽

Lassa Virus ◽

Virus Envelope ◽

Glycoprotein Precursor ◽

Alanine Scan ◽

N Terminus ◽

Hydrophobic Amino Acids ◽

Ph Dependent

Lassa virus glycoprotein 2 (LASV GP-2) belongs to the class I fusion protein family. Its N terminus contains two stretches of highly conserved hydrophobic amino acids (residues 260–266 and 276–298) that have been proposed as N-terminal or internal fusion peptide segments (N-FPS, I-FPS) by analogy with similar sequences of other viral glycoproteins or based on experimental data obtained with synthetic peptides, respectively. By using a pH-dependent, recombinant LASV glycoprotein mediated cell–cell fusion assay and a retroviral pseudotype infectivity assay, an alanine scan of all hydrophobic amino acids within both proposed FPSs was performed. Fusogenicity and infectivity were correlated, both requiring correct processing of the glycoprotein precursor. Most point mutations in either FPS accounted for reduced or abolished fusion or infection, respectively. Some mutations also had an effect on pre-fusion steps of virus entry, possibly by inducing structural changes in the glycoprotein. The data demonstrate that several amino acids from both hydrophobic regions of the N terminus, some of which (W264, G277, Y278 and L280) are 100 % conserved in all arenaviruses, are involved in fusogenicity and infectivity of LASV GP-2.

Download Full-text