Semiquantitation of human chemokine mRNA levels with a newly constructed multispecific competitor fragment

Chemokines are a group of inducible, locally acting proinflammatory cytokin es which have been implicated in the pathogenesis of a variety of diseases. Important members of the group include monocyte chemoattractant protein (M CP)-1, -2, -3, macrophage inhibitory protein (MIP)-1 alpha, -1 beta and RAN TES (regulated upon activation, normal T expressed and secreted). To facili tate further investigation of the human chemokines, we have constructed a n ovel multispecific competitor fragment containing primer binding sites for the CC-chemokines MCP-1, MCP-2, MCP-3, MIP-1 alpha, MIP-1 beta and RANTES, the CXC-chemokines MIP-2 alpha, MIP-2 beta as well as for the housekeeping gene beta-actin. Using this competitor fragment we can demonstrate reliable semiquantitation of reverse transcribed chemokine mRNAs. The assay should be useful for further studies, in particular for the semiquantitation of ch emokine mRNA species from small cell or tissue specimens. (C) 1999 Elsevier Science B.V. All rights reserved.