In vivo microdialysis sampling of cytokines produced in mice given bacterial lipopolysaccharide.
Cytokines are proteins that mediate communication between cells of the immune system as well as certain other non-immune host cells. These proteins are produced by many cell types and they mediate immune and inflammatory responses. However, the direct site analysis of these critical proteins is hampered by the lack of site-specific tools available for such direct measurements. In this study, both in vitro and in vivo microdialysis sampling of different cytokines (tumor necrosis factor-alpha [TNF-alpha], interferon-gamma [IFN-gamma], interleukin-6 [IL-6], IL-12p70, and macrophage chemoattractant protein-1 [MCP-1]) was performed. A mouse model of bacterial lipopolysaccharide (LPS) administration and response pattern was used for in vivo studies. Three cytokines, TNF-alpha, IL-6, and MCP-1 were quantified in the serum from mice given LPS. In vivo studies demonstrated the ability to monitor increasing levels of these cytokines (TNF-alpha, IL-6, and MCP-1) via microdialysis probes placed in the peritoneal cavity of mice given LPS. All three cytokines were quantified simultaneously in 15 muL of dialysate using a multiplexed bead-based immunoassay for flow cytometry. The detected dialysate cytokine concentrations varied between 200 pg/mL and 1500 pg/mL for TNF-alpha, between 600 pg/mL and 3000 pg/mL for MCP-1, and between 2700 pg/mL and more than 5000 pg/mL for IL-6. The detected serum cytokine concentrations ranged from 5700 pg/mL to 35,000 pg/mL for TNF-alpha, from 40,000 pg/mL to 65,000 pg/mL for MCP-1, and greater than than 100,000 pg/mL for IL-6. This work demonstrates that microdialysis sampling can be used in vivo to collect temporal profiles of cytokine production.
Ao
Xiaoping
X
Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Cogswell Laboratories, 110 8th Street, Troy, NY 12180-3590, USA.
Rotundo
Robert F
RF
Loegering
Daniel J
DJ
Stenken
Julie A
JA
D000818:Animals; D001202:Ascitic Fluid / Q000737:chemistry; D018932:Chemokine CCL2 / Q000032:analysis; D016207:Cytokines / Q000032:analysis* / Q000096:biosynthesis / Q000097:blood; D005434:Flow Cytometry / Q000379:methods; D007371:Interferon-gamma / Q000032:analysis; D018664:Interleukin-12 / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008828:Microbiological Techniques / Q000295:instrumentation; D017551:Microdialysis / Q000295:instrumentation / Q000379:methods*; D021122:Protein Subunits / Q000032:analysis; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D016207:Cytokines; D008070:Lipopolysaccharides; D021122:Protein Subunits; D014409:Tumor Necrosis Factor-alpha; D018664:Interleukin-12; D007371:Interferon-gamma