Eukaryotic expression of the broad-spectrum chemokine receptor antagonist vMIP-II and its effects on T-cell function in vitro and in vivo

: Pro‐inflammatory chemokines and their receptors exhibit elementary functions in cell migration and in Th1‐driven inflammatory conditions. One therapeutic strategy to prevent accumulation of pro‐inflammatory immune cells is the use of specific chemokine receptor antagonists. An interesting and promising candidate in this context is the viral antagonist MIP‐II (vMIP‐II) that acts on a broad spectrum of chemokine receptors. To study the in vitro and in vivo effects of vMIP‐II on pro‐inflammatory chemokine receptor function, we further characterized an ovalbumin‐specific murine central memory Th1IF12 clone by using RT‐PCR, cDNA array and cytometry. Using in vitro chemotaxis assays we show that eukaryotically generated vMIP‐II strongly inhibited migration of CCL2‐ or CCL5‐stimulated Th1 IF12 cells. Using intravital microscopy, we observed that CCL5 induced rolling of Th1 cells in the ear vasculature of C57Bl/6 mice. Pre‐treatment with vMIP‐II significantly reduced CCL5‐induced rolling of Th1 cells to basal levels, indicating, that vMIP‐II is also active in vivo (proportion of rolling cells: 19.4 ± 3.8%, 39.8 ± 2.9% and 26.1 ± 3.2%). In addition, investigating the anti‐inflammatory action of vMIP‐II in adoptive transfer of immunity and dinitrofluorobenzene‐induced cutaneous hypersensitivity reaction using C57Bl/6 mice, we show a direct inhibitory effect of vMIP‐II on the sensitization phase [Δ ear swelling 62 and 37 cm × 10−3 for controls and vMIP‐II treated mice (2.5 mg/kg), respectively] and effector phase (Δ ear swelling 14.8 and 3.6 cm × 10−3 for controls and vMIP‐II treated mice (2.5 mg/kg), respectively) of cutaneous hypersensitivity. These data indicate that vMIP‐II is a promising agent to interfere with chronic inflammatory (skin) diseases.