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J Biol Chem, Vol. 274, Issue 45, 32478-32485, November 5, 1999
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From the Modification of the amino terminus of regulated
on activated normal T-cell expressed (RANTES) has been shown to have a
significant effect on biological activity and produces proteins with
antagonist properties. Two amino-terminally modified RANTES proteins,
Met-RANTES and aminooxypentane-RANTES (AOP-RANTES), exhibit
differential inhibitory properties on both monocyte and eosinophil
chemotaxis. We have investigated their binding properties as well as
their ability to activate the RANTES receptors CCR1, CCR3, and CCR5 in
cell lines overexpressing these receptors. We show that Met-RANTES has
weak activity in eliciting a calcium response in Chinese hamster ovary
cells expressing CCR1, CCR3, and CCR5, whereas AOP-RANTES has full
agonist activity on CCR5 but is less effective on CCR3 and CCR1. Their
ability to induce chemotaxis of the murine pre-B lymphoma cell line,
L1.2, transfected with the same receptors, consolidates these results.
Monocytes have detectable mRNA for CCR1, CCR2, CCR3, CCR4, and
CCR5, and they respond to the ligands for these receptors in chemotaxis
but not always in calcium mobilization. AOP-RANTES does not induce
calcium mobilization in circulating monocytes but is able to do so as
these cells acquire the macrophage phenotype, which coincides with a
concomitant up-regulation of CCR5. We have also tested the ability of
both modified proteins to induce chemotaxis of freshly isolated
monocytes and eosinophils. Cells from most donors do not respond, but
occasionally cells from a particular donor do respond, particularly to
AOP-RANTES. We therefore hypothesize that the occasional activity of
AOP-RANTES to induce leukocyte chemotaxis is due to donor to donor
variation of receptor expression.
Serono Pharmaceutical Research Institute,
14 Chemin des Aulx, 1228 Plan-les-Ouates, Geneva, Switzerland,
¶ LeukoSite Inc., Cambridge, Massachusetts 02142, the
** Department of Dermatology and Allergology, University of Kiel,
Kiel, Germany, and Département de Biochimie
Médicale, Centre Medical Universitaire, Geneva
1228, Switzerland
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