Nci h1299 cells

NCI-H1299 cells (ATCC) were grown in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin‒streptomycin. The SVA ZJ-2015 strain was preserved in our laboratory. Briefly, NCI-H1299 cells infected with SVA ZJ-2015 were collected and subjected to three freeze‒thaw cycles. Cellular debris was removed by centrifugation at 6000 × g for 30 min at 4 °C. The virus was then precipitated with 8% polyethylene glycol (PEG 8000) and 0.5 M NaCl for 16 h at 4 °C. The precipitated virus was further centrifuged by a 10–50% sucrose density gradient. Subsequently, the virus band was taken up for desucrose treatment, and the precipitate was resuspended in PBS (pH = 7.4) and stored at − 80°C. Purified virus was used for the preparation of infectious and immune sera.

Human umbilical vein endothelial cells (HUVEC), Human microvascular endothelial cell line (HMEC-1), and Human brain vascular pericytes (HBVPs) were obtained from ScienCell Research Laboratories. HUVECs and HMEC-1 cells were cultured in ECM, and HBVPs were cultured in PM. The human non-small lung cancer cell line NCI-H1299 cells (it is isolated from a NSCLC patient with lymph node metastasis who have received prior radiation therapy and has a homozygous partial deletion of the p53 protein) were purchased from American Type Culture Collection (ATCC, Manassas, Virginia) and cultured with DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. All these cells were maintained in humidified atmosphere containing 5% CO₂ at 37 °C. We showed that all these cells have no cross contamination of other human cell lines using the STR Multi-Amplification Kit (Microreader 21 ID System).

Human bronchial epithelial cells (HBEpiC), human lung cancer cell lines (A549 cells and NCI‐H1299 cells) and human myeloid leukaemia mononuclear cells (THP‐1) were obtained from American Type Culture Collection (ATCC). NCI‐H1299 and THP‐1 were cultured in RPMI 1640 supplemented with 10% FBS (Thermo Scientific, Waltham, Massachusetts, USA). HBEpiC and A549 were cultured in HycloneDME‐F12 supplemented with 10% FBS. All cells were cultured in 5% CO2 at 37°C. LPS and recombinant protein IL‐4 were from Beyotime Biotechnology (Nanjing, China).

The human colon cancer HT29 cells, hepatocellular carcinoma Hep3B cells, breast cancer MCF-7 cells, large cell lung cancer NCI-H460 cells, lymph node metastasized lung cancer NCI-H1299 cells, normal human bronchial epithelial BEAS-2B cells, and murine Lewis lung carcinoma (LLC) cells were obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA). The HT29, Hep3B, MCF-7, BEAS-2B, and LLC cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Welgene, Daegu, Korea) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Sigma-Aldrich) and 1% penicillin/streptomycin (Gibco, Rockville, MD, USA). The NCI-H460 and NCI-H1299 cells were cultured with Roswell Park Memorial Institute 1640 (RPMI1640; Welgene) containing 10% heat-inactivated FBS and 1% penicillin/streptomycin. All cells were cultured at 37 °C in an atmosphere containing 5% CO₂/air.