SPECIFIC ELISAS FOR THE DETECTION OF HUMAN MACROPHAGE INFLAMMATORY PROTEIN-1-ALPHA AND PROTEIN-1-BETA

Mononuclear cell elicitation has gained renewed interest with the discovery of a supergene family of small polypeptide chemotactic cytokines (<10 kD). These chemotactic cytokines have been divided into the C-X-C and C-C chemokine families depending upon whether the first two conserved cysteine amino acid residues are separated by one amino acid or are in juxtaposition, respectively. A salient feature of the C-C chemokine family is their ability to induce both monocyte and lymphocyte chemotaxis. Although monocyte and lymphocyte migration in vitro is measured in chemotactic bioassays, this technique often fails to determine the specific quantitative contribution of a chemotaxin to a biological specimen. Our laboratory has developed two sensitive and specific sandwich ELISAs for the detection of macrophage inflammatory protein-1 alpha and beta (MIP-1alpha and MIP-1beta). The lower threshold for detection of both MIP-1alpha and MIP-1beta was 100 pg/ml, and both of these ELISAs were efficacious for the detection of MIP-1alpha and MIP-1beta in conditioned media from pulmonary fibroblasts, monocytes, neutrophils, and a pulmonary epithelial cell line. The development of these ELISAs will allow the measurement of MIP-1alpha and MEP-1beta from biologically relevant fluids and ascertain whether these two C-C chemokines are present in disease.