BHK-21 Cell Clones Differ in Chikungunya Virus Infection and MXRA8 Receptor Expression

Abstract

Baby hamster kidney-21 (BHK-21) cells are widely used to propagate and study many animal viruses using infection and transfection techniques. Among various BHK-21 cell clones, the fibroblast-like BHK-21/C-13 line and the epithelial-like BHK-21/WI-2 line are commonly used cell clones for alphavirus research. Here we report that BHK-21/WI-2 cells were significantly less susceptible to primary infection by the alphavirus chikungunya virus (CHIKV) than were BHK-21/C-13 cells. The electroporation efficiency of alphavirus RNA into BHK-21/WI-2 was also lower than that of BHK-21/C-13. The growth of CHIKV was decreased in BHK-21/WI-2 compared to BHK-21/C-13, while primary infection and growth of the alphavirus Sindbis virus (SINV) were equivalent in the two cell lines. Our results suggested that CHIKV entry could be compromised in BHK-21/WI-2. Indeed, we found that the mRNA level of the CHIKV receptor MXRA8 in BHK-21/WI-2 cells was much lower than that in BHK-21/C-13 cells, and exogenous expression of either human MXRA8 or hamster MXRA8 rescued CHIKV infection. Our results affirm the importance of the MXRA8 receptor for CHIKV infection, and document differences in its expression in two clonal cell lines derived from the original BHK-21 cell cultures. Our results also indicate that CHIKV propagation and entry studies in BHK-21 cells will be significantly more efficient in BHK-21/C-13 than in BHK-21/WI-2 cells.

Understanding the Biology and Immune Pathogenesis of Chikungunya Virus Infection for Diagnostic and Vaccine Development

“…In Mxra8-deficient mice, decreased CHIKV infection and CHIKV-induced joint swelling were observed [ 15 ]. In line with these findings, ectopic expression of Mxra8 in vitro in various cell lines and in vivo in Drosophila led to enhanced CHIKV invasion [ 15 , 16 , 18 ].…”

“…Conversely, removal of this insertion resulted in enhanced susceptibility of Bovinae to CHIKV invasion [ 17 ]. In vitro, cell lines with low expression levels of Mxra8 were less susceptible to CHIKV infection [ 18 ]. In Mxra8-deficient mice, decreased CHIKV infection and CHIKV-induced joint swelling were observed [ 15 ].…”

A single-point mutation in the rubella virus E1 glycoprotein promotes rescue of recombinant vesicular stomatitis virus

“…Vero cells were maintained in high glucose Dulbecco's Modified Eagle's Medium (DMEM) containing 1 mM sodium pyruvate and supplemented with 10% fetal bovine serum (FBS), 1% L-glutamine, and 1% penicillin-streptomycin (Pen-Strep). BHK-21/WI-2 and BHK-21/C-13 cells were cultured in high glucose DMEM supplemented with 1% L-glutamine, 1% Pen-Strep, and 10% tryptose phosphate broth and either 5% (BHK-21/WI-2) or 10% (BHK-21/C-13 cells) FBS ( 60 ). U-2 OS cells were cultured in modified McCoy’s 5A medium supplemented with 10% FBS and 1% Pen-Strep.…”

“…1 × 10 4 cells were analyzed for each sample using a BD LSR-II analyzer (BD Biosciences, San Jose, CA, USA) in the Einstein flow cytometry core. Mock-infected cells stained as above were used to delineate the gates for flow analysis ( 60 ). The flow data were processed using FlowJo 10.2 software.…”