Abstract
BackgroundEnterovirus A 71 (EV71) is a neurotropic virus that may lead to acute flaccid paralysis, encephalitis, cardiopulmonary failure or even death. No vaccine and defensive drug controlling EV71 is currently available, novel and efficient antiviral drug or vaccine is therefore urgently needed. 3Dpol (RNA-dependent RNA polymerase (RdRp)) has been an important target for anti-EV71 drug development.MethodsA panel of monoclonal IgG antibodies (mAbs) against EV71 3Dpol were generated by traditional cell fusion methods. And the antibody affinity and specificity to EV71 3Dpol were evaluated by Enzyme-linked Immunosorbent Assay (ELISA), Indirect Fluorescent Assay (IFA) and Western blotting. Antiviral activities of these antibodies were also determined in vitro and in vivo.ResultsTwo mAbs towards EV71 3Dpol were able to effectively suppress EV71 replication in Vero-1008 cell when intracellarly delivered. And they also dampened the RNA polymerase activity of 3Dpol in vitro. More importantly, these mAbs provided partial protection in EV71-challenged neonatal murine challenge model.ConclusionsThese results showed that two of mAbs against EV71 3Dpol inhibited EV71 replication and could be utilized as promising therapeutic drug candidate.
Highlights
Enterovirus A 71 (EV71) is a neurotropic virus that may lead to acute flaccid paralysis, encephalitis, cardiopulmonary failure or even death
We achieved three Monoclonal antibody (mAb) 3A12, 2A10 and 7A6G1, and the property of each mAb was summarized in Table 1. 3A12 and 7A6G1 are IgG1 subtype with typical heavy chains about 50 kD and light chains about 27 kD, while 2A10 is IgG2a subtype with typical heavy chains about 50 kD and light chains about 24 kD (Fig. 1a)
Specificities of the mAbs were identified with EV71-infected Vero-1008 cells by Indirect Fluorescent Assay (IFA) using the mAbs as primary antibodies
Summary
Enterovirus A 71 (EV71) is a neurotropic virus that may lead to acute flaccid paralysis, encephalitis, cardiopulmonary failure or even death. The antibody affinity and specificity to EV71 3Dpol were evaluated by Enzyme-linked Immunosorbent Assay (ELISA), Indirect Fluorescent Assay (IFA) and Western blotting. Antiviral activities of these antibodies were determined in vitro and in vivo. Results: Two mAbs towards EV71 3Dpol were able to effectively suppress EV71 replication in Vero-1008 cell when intracellarly delivered. They dampened the RNA polymerase activity of 3Dpol in vitro. These mAbs provided partial protection in EV71-challenged neonatal murine challenge model.
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