Influence of High Hydrostatic Pressure on Epitope Mapping of Tobacco Mosaic Virus Coat Protein

Abstract

In this study, we investigated the effect of high hydrostatic pressure (HHP) on tobacco mosaic virus (TMV), a model virus in immunology and one of the most studied viruses to date. Exposure to HHP significantly altered the recognition epitopes when compared to sera from mice immunized with native virus. These alterations were studied further by combining HHP with urea or low temperature and then inoculating the altered virions into Balb-C mice. The antibody titers and cross-reactivity of the resulting sera were determined by ELISA. The antigenicity of the viral particles was maintained, as assessed by using polyclonal antibodies against native virus. The antigenicity of canonical epitopes was maintained, although binding intensities varied among the treatments. The patterns of recognition determined by epitope mapping were cross checked with the prediction algorithms for the TMVcp amino acid sequence to infer which alterations had occurred. These findings suggest that different cleavage sites were exposed after the treatments and this was confirmed by epitope mapping using sera from mice immunized with virus previously exposed to HHP.

Inhibitor and Substrate Binding Induced Stability of HIV-1 Protease against Sequential Dissociation and Unfolding Revealed by High Pressure Spectroscopy and Kinetics

“…The structural changes are most often studied by variety of spectroscopic methods. However, structural distortions can result also in different set of antibodies obtained from an experimental animal after immunization by antigen treated by high pressure [ 25 ]. High pressure methods can be used also for preparative purposes, e.g.…”

Porcine parvovirus VP1/VP2 on a time series epitope mapping: exploring the effects of high hydrostatic pressure on the immune recognition of antigens

“…The efficacy of HHP for inactivating a variety of foodborne pathogenic microorganisms is well-established, and some of these microorganisms have been demonstrated to retain immunogenic properties, a finding which suggests that HHP may have an application in vaccine development. Indeed, pressure can result in virus inactivation while preserving immunogenic properties [11,12,14,21]. The forces governing viral assembly and disassembly rely on both the microenvironment and the protein-protein interaction network produced by their genetic codes [54].…”

“…Ten 21-day-old specific-pathogen-free (SPF) male pigs from a terminal cross of Landrace and Large White lineages were divided into five groups of two pigs each: N (native PPV), P (PPV treated with 350 MPa at 25°C), P-18 (PPV treated with 350 MPa at − 18°C), V (commercial inactivated PPV vaccine, Farrow sure B, Pfizer) and NC (negative control). Pigs were vaccinated intramuscularly on day 0 (D0), D14, D28 and D38 according to each HHP protocol, as previously described [21]. Pigs in the NC group received only saline solution.…”

Porcine Parvovirus VP1/VP2 on a Time Series Epitope Mapping: exploring the effects of high hydrostatic pressure on the immune recognition of antigens

“…The efficacy of HHP for inactivating a variety of foodborne pathogenic microorganisms is well established, and some of these microorganisms have been demonstrated to retain immunogenic properties, a finding which suggests that HHP may have an application in vaccine development. Indeed, pressure can result in virus inactivation while preserving immunogenic properties 11,12,14,50 .…”

“…4 and (NC) (negative control). Pigs were vaccinated intramuscularly on day 0 (D0), D14, D28 and D38 according to each HHP protocol, as previously described 50 . At necropsy following euthanasia (D72), samples of lung, heart, liver, spleen, kidney and mesenteric lymph node were collected and stored at -70 °C until analyzed by nested-PCR.…”