Antigen-induced cytokine and chemokine release test for tuberculosis infection using adsorption of stimulated whole blood on filter paper and multiplex analysis.

Author(s): Skogstrand K, Thysen AH, Jørgensen CS, Rasmussen EM, Andersen AB, Lillebaek T, Hougaard DM, Houen G

Publication: Scand J Clin Lab Invest, 2012, Vol. 72, Page 204-11

PubMed ID: 22283828 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of conditions of LPS stimulation of blood and preservation of LPS-stimulated blood as DBS or frozen plasma on the levels of 21 inflammatory markers.

   Summary of Findings:

   Inflammatory markers increased when blood was incubated with LPS, and the rate of increase was larger at 37 degrees C than at room temperature; however, after 24 h a greater change in IFN-gamma was observed at room temperature than when incubated at 37 degrees C. Increases in inflammatory marker levels were generally not found when blood was incubated with saline at room temperature, but small increases were seen after incubation with saline at 37 degrees C. While increases in inflammatory marker levels were noted when specimens were stored, after LPS stimulation, as frozen plasma or as DBS, concentrations of IL-6, 8 and 10 were higher in frozen plasma, and IFN-gamma was higher in DBS. Levels of IL-4, IL-8, IL-18, RANTES, MMP-9, IFN-gamma, IL-1 beta, TNF-beta, MCP-1, and TGF-beta were generally lower, while levels of GM-CSF, IL-2, sIL-6Ra, MIP-1 alpha, IL-17, and TNF-alpha were generally higher in frozen plasma than in DBS from unstimulated blood.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Plasma

   Preservative Types

   • Frozen
   • Other Preservative

   Diagnoses:

   • Not specified
   • Normal
   • Other diagnoses

   Platform:

   Analyte	Technology Platform
   Protein	ELISA
   Cell count/volume	Lymphocyte proliferation assay
   Cell count/volume	Light microscopy
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Air-dried Frozen
   Preaquisition	Diagnosis/ patient condition	Tuberculosis
   Analyte Extraction and Purification	Incubation duration/condition	Room temperature 37 degrees C 5 min 2 h 4 h 6 h 24 h With saline With LPS
   ELISA Specific	Targeted peptide/protein	IFN gamma IL-4 IL-6 IL-8 IL-10 IL-18 RANTES MMP-9 IL-1 beta TNF-beta MCP-1 TGF-beta GM-CSF IL-2 SIL6ra MIP-1 alpha IL-17 TNF-alpha

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