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 disc overy 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 cons erved cysteine amino acid residues are separated by one amino acid or are in juxtaposition, respectively. A salient feature of the C-C chemo kine family is their ability to induce both monocyte and lymphocyte ch emotaxis. Although monocyte and lymphocyte migration in vitro is measu red 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 san dwich ELISAs for the detection of macrophage inflammatory protein-1 al pha and beta (MIP-1alpha and MIP-1beta). The lower threshold for detec tion 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, neutrophi ls, and a pulmonary epithelial cell line. The development of these ELI SAs will allow the measurement of MIP-1alpha and MEP-1beta from biolog ically relevant fluids and ascertain whether these two C-C chemokines are present in disease.