Cytokine analysis
Cytokines, tumor necrosis
REGRESSION, TUMOR NECROSIS
The concentrations of 27 cytokines were measured by Luminex in cervicovaginal exudates using the Bio-Plex Pro Human 27-plex cytokine assay (Bio-Rad). These included interleukin (IL)-1RA, IL-1β, IL-2, -4, -5, -6, -7, -8, -9, -10, -12(p70), -13, -15, -17, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, Eotaxin, Granulocyte-macrophage colony-stimulating factor (GM-CSF), Granulocyte colony-stimulating factor (G-CSF), platelet-derived growth factor (PDGF)-BB, fibroblast growth factor-basic (FGF-basic), vascular endothelial GF (VEGF), IFN-γ-inducible-protein (IP)-10, macrophage chemotactic protein (MCP)-1, Regulated on Activation normal T cell expressed and activated (RANTES), macrophage inflammatory protein (MIP)-1α, and MIP-1β. Data were collected using Bio-plex manager software version 4 (Bio-Rad). A 5 Parameter Logistic (5 PL) regression formula was used to calculate sample concentrations from the standard curves. If the concentration of a cytokine fell outside of the limits of detection for a given sample, then that value was not used in statistical analysis. If any cytokine was undetectable in >40% of samples assayed, it was excluded from analyses beyond its initial description. Specimens from five participants were included across all plates (inter-plate controls). In addition, samples from five participants were duplicated on each set of plates (intra-plate controls) for quality control measures. Spearman’s rank test was used to measure intra-assay and inter-assay correlation coefficients to determine assay reliability and reproducibility. Cytokines were excluded from this analysis if the inter-assay correlation <0.8 (which included IL-12(p70)).