Bhk 21 cells ccl 10

MuPyV.A2 was prepared in baby mouse kidney cells as described [82 (link)]. Mice were infected intracerebrally (i.c.) with 3x10⁵ PFU MuPyV.A2 in 30 μL as described [34 (link)]. For rechallenge, mice were inoculated i.c. with 3x10⁵ PFU MuPyV.A2 in 30 μl at day 0, re-inoculated i.c. with 3x10⁵ PFU MuPyV.A2 in 30 μl or vehicle at day 30 p.i., then euthanized 4–5 days later. Recombinant VSV expressing the MuPyV D^bLT359 epitope (VSV.LT359) was grown and titered on BHK-21 cells (CCL-10; ATCC, Manassas VA) [40 (link)]. Mice were infected intranasally with 5x10⁴ PFU rVSV-LT359 diluted in PBS.

BHK-21 cells (CCL-10) were obtained from ATCC (USA) and cultured in modified essential eagle medium (Sigma-Aldrich, USA) supplemented with 10% foetal bovine serum (FBS), 2 mM L-Glutamine, 100 U penicillin & 0.1 mg/ml streptomycin (Sigma-Aldrich). CEF cells were obtained from the Institute for Animal Health (Compton, UK) and cultured in Dulbecco's modified eagle medium (Sigma-Aldrich), supplemented as above. SW13 (accession number 87031801) and Vero E6 (accession number 85020206) cells were obtained from the European Collection of Cell Cultures, UK. Cultures were maintained in Leibovitz's L-15 medium containing Glutamax (Life Technologies, UK) supplemented with 10% FBS.

The Bangladeshi strain of the Nipah virus (NIV) was obtained from the Viral Special Pathogens Branch of the CDC, was originally isolated from a fatal human infection in Bangladesh in 2004, and was passaged in Vero E6 cells a total of three times [16 (link)]. Cedar virus (CedPV) was obtained from CSIRO (Geelong, Australia) and was isolated from bat urine inoculated onto primary bat kidney cells, followed by passaging on Vero E6 cells (ATCC) [7 (link)]. Both viruses were propagated on Vero E6 cells at RML in Dulbecco’s minimal essential medium (DMEM) (Sigma) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 50 IU/mL penicillin, and 50 µg/mL streptomycin (Life Technologies, Carlsbad, CA, USA).
BHK-21 cells (CCL-10, ATCC, Manassas VA, USA) were propagated in 24-well tissue culture plates and inoculated with CedPV or NiV (or mock) with a MOI of 1.0 or 0.1 for 1 h, washed twice in DMEM, and incubated at 37 °C, 5% CO₂ for the indicated amount of time.
Human lung blood microvascular endothelial cells (LBMVEC) (CC-2527, Lonza, Walkersville MD, USA) were maintained in an EGM-2 medium (Lonza). The cells were seeded in 24-well plates one day prior to inoculation with CedPV or NiV with a MOI of 0.1. At the indicated time points, supernatants were collected for virus quantitation and the cells were lysed in buffer RLT (Qiagen) to quantitate viral or cellular RNA.