Mab478

To neutralize CCL5, CXCL9, and CXCL10 in vivo, anti-mouse CCL5 (R&D Systems, MAB478, 50 μg per mouse), anti-mouse CXCL9 (R&D Systems, AF-492-NA, 50 μg per mouse), anti-mouse CXCL10 (R&D Systems, MAB466, 50 μg per mouse) or a mixture of these antibodies were intraperitoneally injected twice weekly.
For CD40/CD40L signal blockade in vivo, an anti-mouse CD40L monoclonal antibody (BioXCell, MR-1, 300 μg per mouse) was intraperitoneally administered twice weekly.
To deplete CD4+ T cells or CD8+ T cells, mice were injected intraperitoneally with 400 μg of an anti-mouse CD4 antibody (BioXCell, GK1.5) or an anti-mouse CD8 antibody (BioXCell, YST-169.4) every 3 days.

Cohorts of mice were treated via ic injection with Met-RANTES (1 μg/mouse; Bachem, Basel, Switzerland), vehicle, anti-RANTES mAb (MAB478), or an IgG2a isotype-matched control mAb (10 μg/mouse; R&D Systems) on days 2 to 8 after ic infection with TBEV WH2012 (10³ TCID₅₀s). Met-RANTES is a recombinant RANTES analog, in which the initiating methionine residue is retained after expression in Escherichia coli cells, resulting in a potent antagonist of the murine RANTES receptors CC chemokine receptor (CCR)5 and CCR1 [25 (link), 26 (link)]. The chosen RANTES-neutralizing mAb reacts with murine and human RANTES and no other identified cytokine or chemokine [27 (link), 28 (link)]. Mice were monitored daily for signs of neurological disease and survival over a period of 14 days. The survival curves were compared using Kaplan-Meier tests. Both brain virus titers and mice survival rates were estimated at the time points indicated below. The severity of inflammation was determined by staining sections of paraffin-embedded brain tissue with HE by day 8 post infection (p.i.), as after this time, the efficacy of this treatment may decline due to the decay of corresponding molecules within the mouse brain.