稳定表达抗口蹄疫病毒IgA抗体细胞系的建立与应用OA

This study aims to establish a CHO-K1 cell line that can stably express immunoglobulin A(IgA)antibodies against foot-and-mouth disease virus(FMDV),which will lay a foundation for future research on the antiviral properties of IgA and secretory IgA(sIgA).Here,the heavy chain gene and the light chain gene of FMDV serotype O-and serotype A-specific IgA antibodies were amplified separately based on previously constructed expression plasmids encoding the heavy chain and light chains of anti-FMDV serotypes O and A IgA and cloned into the lentiviral vectors pLVX-Puro and pLVX-Hygro to generate lentiviral plasmids 8GK-Puro and 8GH-Hygro.The resulting heavy-chain and light-chain lentiviral plasmids as well as the envelope plasmid pMD2.G and the packaging plasmid pCMV-dR8.74 were co-transfected into HEK-293T cells to package lentiviruses.The harvested lentiviruses were used to infect CHO-K1 cells and the transfected cells were firstly selected with puromycin and hygromycin B and then subjected to limited dilution to isolate a stable CHO-K1 cell line expressing IgA antibodies.The selected cell line was analyzed at the genetic and protein levels by RT-PCR,indirect immunofluorescence assay,and Western-blot.The results demonstrated that the genes encoding the heavy and light chains of the IgA antibody were successfully integrated into the cell line's genome and the cell line can stable expression heavy and light chains of the IgA.This study successfully established a stable cell line expressing anti-serotype O and serotype A FMDV IgA antibodies,which will provide a basis for antiviral studies on anti-FMDV IgA and sIgA as well as development of anti-FMDV drugs in the future.