Expression and self-assembly of recombinant capsid protein from the antigenically distinct Hawaii human calicivirus

Abstract

The Norwalk and Hawaii viruses are antigenically distinct members of the family Caliciviridae and are considered to be important etiologic agents of epidemic gastroenteritis, with most studies focusing on the role of Norwalk virus. To further investigate the importance of Hawaii virus, Hawaii virus-like particles (VLPs) were produced by expression of its capsid protein in the baculovirus system and these VLPs were used as the antigen in an enzyme-linked immunosorbent assay that was efficient in the detection of a serologic response to Hawaii virus. The ready availability of Hawaii VLPs should enable larger-scale epidemiological studies to further elucidate the importance of this agent.

Expression and Self-Assembly in Baculovirus of Porcine Enteric Calicivirus Capsids into Virus-Like Particles and Their Use in an Enzyme-Linked Immunosorbent Assay for Antibody Detection in Swine

“…It has many advantages over other expression systems: (i) high expression efficacy, (ii) eukaryotic posttranslational modifications, (iii) preservation of biological properties of recombinant proteins, and (iv) self-assembly of viral capsid proteins into VLPs for both RNA and DNA viruses (3,5,13,16,21,22,26,27). The capsid proteins of NV, MxV (a Snow Mountain Agent-like virus), Lordsdale virus, Toronto virus, Hawaii virus, and RHDV were expressed efficiently in baculovirus, and the recombinant capsids self-assembled into VLPs that were antigenically and morphologically similar to their respective native viruses (5,13,21,22,26,27). The recombinant capsids of Sapporo virus and the SLV Houston/86/US and Houston/90/US strains were also expressed in baculovirus (24,32), but the yields of VLPs were relatively low and the VLPs tended to be unstable at pHs above 7 (32).…”

“…Because HuCV are uncultivable in cell culture and are genetically diverse, epidemiological investigations of human infections were impeded until recombinant calicivirus capsids which assembled into virus-like particles (VLPs) were produced and used to develop serologic diagnostic assays (12,19,22,23). The baculovirus expression system has proven very efficient for the production of recombinant capsids of NLVs such as Norwalk virus (NV), Mexico virus (MxV), Toronto virus, Hawaii virus, Grimsby virus, and also the rabbit hemorrhagic disease virus (RHDV), a lagovirus causing systemic hemorrhage and liver necrosis in rabbits (5,13,16,21,22,26,27). Interestingly, the recombinant capsid proteins expressed in baculovirus selfassembled into VLPs which were morphologically and antigenically similar to native viruses.…”

Self-Assembly of the Recombinant Capsid Protein of a Bovine Norovirus (BoNV) into Virus-Like Particles and Evaluation of Cross-Reactivity of BoNV with Human Noroviruses

“…Virus-like particles (VLPs) of noroviruses and sapoviruses can be generated by a baculovirus expression system and are morphologically and antigenically similar to native virus. They have been used to produce the antigens and antibodies needed for diagnostic assays (18,19,22,28,29,33). Recently, VLPs of Bo/Jena/80/UK (GIII, genotype 1 [GIII/1]) were reported (12).…”

“…An enzyme immunoassay with VLPs of Norwalk virus as the antigen was used as a serotyping assay for the detection of antibodies or antigens to the prototype Norwalk virus as well as those to other human noroviruses (18,19,22,28,29). In addition, VLPs can be used to determine the antigenic relationships among enteric caliciviruses and to study the relationships between different serotypes or genotypes (3).…”

Antiviral strategies to control calicivirus infections

“…Virus-like particles that mimic structurally and antigenically the native capsid protein have been extensively used to explore the function and reactivity of noroviruses and sapoviruses (Guo et al, 2001b;Hansman et al, 2005aHansman et al, , 2006Jiang et al, 1999b;Oka et al, 2006). VLPs are produced to relatively high yields in baculovirus recombinant expression systems or in the human endothelial kidney cell line HEK-293T (Taube et al, 2005) assembling in the absence of the VP2, although the VP2 seems to enhance VLPs stability (Green et al, 1997;Jiang et al, 1992). Furthermore, feeding of volunteers and laboratory animals with norovirus virus-like particles generated in tomato plants (Huang et al, 2005b) or potato plants (Tacket et al, 2000) led to an immune response (IgG and IgA) with variable intensity, depending on the adjuvant used.…”