A Genetically Engineered Attenuated Coxsackievirus B3 Strain Protects Mice against Lethal Infection

Abstract

Coxsackievirus B3 (CVB3) is a common human pathogen that is endemic throughout the world. There is currently no vaccine available, although the virus is known to be highly lethal to newborns and has been associated with heart disease and pancreatitis in older children and adults. Previously, we showed that the virulence of CVB3 is reduced by a lysine-to-arginine substitution in the capsid protein VP2 (K2168R) or a glutamic acid-to-glycine substitution in VP3 (E3060G). In this report, we show that the double mutant virus CVB3(KR/EG) displays additional attenuation, particularly for the pancreas, in A/J mice. In addition, two other attenuating mutations have been identified in the capsid protein VP1. When either the aspartic acid residue D1155 was replaced with glutamic acid or the proline residue P1126 was replaced with methionine, the resulting mutant also possessed an attenuated phenotype. Moreover, when either of these mutations was incorporated into CVB3(KR/EG), the resulting triple mutant viruses, CVB3(KR/EG/DE) and CVB3(KR/EG/ PM), were completely noncardiovirulent and caused only small foci of damage to the pancreas, even at a high dose. Both triple mutants were found to be immunogenic, and a single injection of young A/J mice with either was found to protect them from a subsequent lethal challenge with wild-type CVB3. These findings indicate that the triple mutants could be exploited for the development of a live attenuated vaccine against CVB3.

Coxsackievirus B3 Is an Oncolytic Virus with Immunostimulatory Properties That Is Active against Lung Adenocarcinoma

“…S4B). Although human CAR mRNA is expressed in the heart (44), CVB3 possibly causes severe myocarditis in infants only (45), suggesting its relatively mild side effects in use for adult patients.…”

A Novel Pancreatropic Coxsackievirus Vector Expressing Glucagon-Like Peptide 1 Reduces Hyperglycemia in Streptozotocin-Treated Mice

“…1. This vector is based on an infectious clone of a genetically engineered coxsackievirus B3 (CVB3) strain, CVB3(KR/EG), that we have previously shown to be selectively pancreatropic (7). A multiple cloning site was inserted at the P1/P2 junction and is flanked by two repeated DNA fragments containing the coding region for the 2Apro viral protease cleavage site.…”

“…In previous research on coxsackieviruses, the extraordinarily strong tissue tropism of group B coxsackieviruses (CVBs) for pancreatic tissues (both exocrine and islets) has been demonstrated (7,19,35,48). While CVBs also target heart tissue (12, 39), we have developed a unique pancreatropic strain, constructed during development of an attenuated coxsackievirus B vaccine candidate for humans (7).…”

“…While CVBs also target heart tissue (12, 39), we have developed a unique pancreatropic strain, constructed during development of an attenuated coxsackievirus B vaccine candidate for humans (7). This strain possesses two mutations that restrict viral infection in the heart and liver, directing virus to pancreatic tissues where it causes foci of infection that are distributed widely throughout the acinar tissue (7). By incorporating a multiple cloning site (MCS) into this CVB strain, we have constructed a viral vector [vCVB(dm)] (v stands for vector, CVB stands for group B coxsackievirus, and dm stands for double mutant) that is specific for pancreas and enables sustained expression of an inserted gene for several days.…”

Mucosal co-immunization with AIM2 enhances protective SIgA response and increases prophylactic efficacy of chitosan-DNA vaccine against coxsackievirus B3-induced myocarditis

“…[4][5][6] So far, there is no efficient vaccines or therapeutic reagents against CVB3-induced myocarditis in clinic. [7][8][9][10][11][12][13] Therefore, there is an obvious need to develop a new and efficient vaccine.…”