Development of a pseudovirus based assay for measuring neutralizing antibodies against coxsackievirus B5

Abstract

Coxsackievirus B5 (CV-B5), an important Coxsackie B virus from genus Enteroviruse within the family Picornaviridae, has also been isolated from Hand, Foot, and Mouth Disease (HFMD) patients, and often associated with neurological manifestations. In this study, we found out that Coxsackievirus B3 (CV-B3) replicon RNA could be encapsidated with CV-B5 capsid to assemble infectious CV-B5 pseudovirus. We then utilized this single round infection system of CV-B5 to develop a neutralizing antibody quantification assay. This pseudovirus neutralization assay showed superiority in biosafety, sensibility, quantitativity, efficiency and high throughput, and would facilitate the epidemiological studies and vaccine development of CV-B5.

A surrogate assay for measuring Coxsackievirus A6 neutralizing antibodies

“…In our previous work, we have proposed that some enteroviruses may share similar determinants in virion assembly. 25 Our findings in this study further suggest that the enteroviruses within the same genotype group are more likely to have encapsidation compatibility with each other.…”

“…26 Previously, we had found that capsid proteins from CV-A16 could package EV-A71 replicon into infectious CV-A16 pseudovirus, and capsid proteins from CV-B5 could package CV-B3 replicon RNA into infectious CV-B5 pseudovirus. 25 In this work, we showed another successful application of the trans-encapsidation strategy to produce infectious CV-A6 pseudovirus. EV-A71 and CV-A16 replicon both could be encapsidated with CV-A6 capsid to produce CV-A6 pseudovirus; however, CV-B3 and CV-B5 replicon RNA could not be encapsidated with CV-A6 capsid.…”

“…To date, we have established pseudovirus luciferase assays (PVLA) for EV-A71, 22 CV-A16, 23 Coxsackievirus B3 (CV-B3), 24 and Coxsackievirus B5 (CV-B5). 25 Our work has shown that these assays can be used as surrogates for the CPE and have superiority in terms of biosafety, sensitivity, and quantitativity. In this study, we successfully established a pseudovirus infection system for CV-A6, and then developed a PVLA method for anti-CV-A6 NtAb detection.…”

“…In our previous work on enteroviruses, we have found out that genetically modified viral replicon RNA can be transencapsidated in cells by viral proteins from its own or some other enteroviruses from the same subgroup. 24,25 So far, we have already established several robust pseudoviruses producing systems for EV-A71, CV-A16, CV-B3 and CV-B5. Adopting the similar strategies, we first built the CV-A6 capsid expressor and subgenomic replicon ( Figure 1A).…”

Development of a pseudovirus-based assay for measuring neutralizing antibodies against Coxsackievirus A10

“…[16][17][18][19] Our research group has already constructed numerous types of pseudoviruses relevant to HFMD, including pseudoviruses of enterovirus 71 (EV-A71), CV-A16, CV-B3, CV-B5, and CV-A6, which were used in the development of a pNT assay. [20][21][22][23][24] Here, we report the successful construction of a CV-A10 pseudovirus, and development of a CV-A10 pNT to detect the titre of anti-CV-A10 NtAb. The method was validated using 67 clinical serum samples, and the performance and safety were compared to those of the traditional cNT method.…”