Rat igg2b isotype control ltf 2

All mouse experiments were performed at OHSU in ABSL3 laboratories in compliance with OHSU IACUC protocols. The small lab animal unit at OHSU is accredited by the Association for the Accreditation and Assessment of Laboratory Animal Care (AALAC) International. Animals were housed in ventilated racks and monitored daily by veterinary staff. C57BL/6J mice were vaccinated as indicated with MCMV delivered intraperitoneally (10⁶ PFU, i.p.), and/or AdV injected intramuscularly in the thigh (10⁸ PFU, i.m.). Mice were challenged with 10³ PFU CHIKV in a 20 μl volume in the footpad (f.p.), or they were challenged (i.m.) with 10³ or 10⁴ PFU in a 20 μl volume in the calf muscle. Footpad measurements were taken with calipers. For T cell depletion experiments, mice were administered T cell depleting antibodies diluted in PBS in a 100 μl volume (i.p.). Vaccinated groups were injected with 300 μg anti-CD4 (GK1.5, BioXCell), 300 μg anti-CD8 (2.43, BioXCell), 300 μg Rat IgG2b Isotype Control (LTF-2, BioXCell), or a combination of 300 μg anti-CD4 and 300 μg anti-CD8. T cell depletions were confirmed by flow cytometry. To confirm T cell depletions, splenocytes were stained with fluorophore-conjugated antibodies specific for mouse CD3, CD4, CD8, and CD19. Fluorescent markers were detected on an LSRII instrument (BD Pharminogen) and data was analyzed using FlowJo (TreeStar).