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 [Delta ear swelling 62 and 37 cm x 10(-3) for controls and vMIP-II treated mice (2.5 mg/kg), respectively] and effector phase (Delta ear swelling 14.8 and 3.6 cm x 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.
Rubant
Simone
S
Department of Dermatology, Clinic of the Johann Wolfgang Goethe University, Frankfurt, Germany. s.rubant@em.uni-frankfurt.de
Ludwig
Ralf J
RJ
Pfeffer
Jeannette
J
Schulze-Johann
Petra
P
Kaufmann
Roland
R
Pfeilschifter
Josef M
JM
Boehncke
Wolf-Henning
WH
Radeke
Heinfried H
HH
D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology*; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D018925:Chemokines / Q000494:pharmacology*; D002633:Chemotaxis; D003877:Dermatitis, Contact; D006801:Humans; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019707:Receptors, Chemokine / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000737:chemistry; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*
D019380:Anti-HIV Agents; D000893:Anti-Inflammatory Agents; D018925:Chemokines; D019707:Receptors, Chemokine; D011994:Recombinant Proteins; C108059:vMIP-II