Epitope-mapping of the glycoprotein from Crimean-Congo hemorrhagic fever virus using a microarray approach

Abstract

Crimean-Congo hemorrhagic fever virus (CCHFV) causes severe acute human disease with lethal outcome. The knowledge about the immune response for this human health threat is highly limited. In this study, we have screened the glycoprotein of CCHFV for novel linear B-cell epitopic regions using a microarray approach. The peptide library consisted of 168 synthesized 20mer peptides with 10 amino acid overlap covering the entire glycoprotein. Using both pooled and individual human sera from survivors of CCHF disease in Turkey five peptide epitopes situated in the mucin-like region and GP 38 (G15-515) and GN G516-1037 region of the glycoprotein were identified as epitopes for a CCHF immune response. An epitope walk of the five peptides revealed a peptide sequence located in the GN region with high specificity and sensitivity. This peptide sequence, and a sequence downstream, reacted also against sera from survivors of CCHF disease in South Africa. The cross reactivity of these peptides with samples from a geographically distinct region where genetically diverse strains of the virus circulate, enabled the identification of unique peptide epitopes from the CCHF glycoprotein that could have application in development of diagnostic tools. In this study clinical samples from geographically distinct regions were used to identify conserved linear epitopic regions of the glycoprotein of CCHF.

Vesicular Stomatitis Virus-Based Vaccine Protects Mice against Crimean-Congo Hemorrhagic Fever

“…It is interesting to note, that a similar motif has been demonstrated by other bunyaviruses 47,48 and has also been demonstrated with other VSV pseudotypes 49 . Structural CCHFV-G C has been shown to be an important protein for CCHFV and contains a putative receptor binding region for mammalian cell surface nucleolin 50 , a class II viral fusion domain 51 , a neutralization epitope 13 , and human linear B-cell epitope sites 52,53 . Our replication competent rVSV vector can enable further exploration of these components at lower containment (BSL-2), without the need for transfections for in trans expression of CCHFV-GPC onto VSVΔG systems.…”

Animal Models for Crimean-Congo Hemorrhagic Fever Human Disease

“…Human IgM and IgG antibody responses can be detected against the glycoproteins (GP38, G N and G C ), and nucleocapsid (N) protein in CCHF survivors [48,49,50]. This includes development of neutralizing antibodies responses for which G C is the only known target [12,51].…”

Purification of Crimean–Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis

“…Although CCHFV remains one of the priority pathogens needing urgent research and development of diagnostics, experimental studies involving live infectious CCHFV are restricted to the highest level biosafety containment laboratories around the world 10 , 11 , 16 . Alternatively, recombinant CCHFV proteins have been used to study their functions or to develop serological diagnostic tools.…”

“…In addition, CCHFV requires the highest level of biocontainment (Biosafety level 4), hampering its handling for experimental studies. Thus, there are only a few laboratories that can handle infectious CCHFV for virus isolation and production of its whole native viral antigens for serological diagnostic assays that are expected to be less affected by genomic diversity of CCHFVs than genetic detection 7 , 8 , 10 , 11 .…”