Biological Evaluation of the MILLIPLEX® Human Cytokine/Chemokine/Growth Factor Panel A for Disease Profiling of 48 Immune Factors

Abstract

Abstract Cytokine, chemokine and growth factor research plays a significant role in achieving a deeper understanding of the immune system and disease. We recently developed a MILLIPLEX® Human Cytokine/Chemokine/Growth Factor Panel A multiplex assay for simultaneous measurement of picogram levels of 48 human immune factors using Luminex® xMAP® technology in small volumes of serum, plasma or cell culture samples. Distinctly different stimulant-dependent cytokine responses were observed for challenged human PBMCs. Both LPS and Con-A induced similarly the secretion of 13 analytes, whereas LPS preferentially induced 10 analytes, including TNFα, IL-1α and IL-1β. Con-A preferentially induced 16 analytes including Th17 cell cytokines IL-17A and IL-17F. Plasma samples, from active (n=20) and benign (n=18) ovarian cancer patients (Dr. Richard Moore, U of Rochester Medical Center) were tested at UPMC. Fourteen analytes were significantly elevated in active patients vs. benign patients including: cytokines TNFα, IL-5, M-CSF; chemokines MCP-3, Fractalkine; Growth Factors FLT-3L, PDGF-BB (p<0.05); cytokines IL-6, IL-18 (p<0.02); cytokines IL-7, IL-27, IL-10, IL-13 (key role in cancer pathogenesis); chemokine MIP-1β (p<0.01). Residual serum and plasma sepsis patient samples (n=16) were compared to those from healthy controls (n=20) and 7 analytes were significantly elevated in sepsis patients: cytokines IL-1RA, IL-6 (p<0.05), IL-7, IL-27, MIP-1β (p<0.02), M-CSF; chemokine MIG (p<0.01). Twelve other analytes trended higher in sepsis samples. Thus, the Human Cytokine/Chemokine/Growth Factor Panel A is a useful tool for profiling 48 immune factors both in cell culture or blood samples for the study of human disease and inflammation.