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Fig. 4 | Arthritis Research & Therapy

Fig. 4

From: Hyaluronidase treatment of synovial fluid is required for accurate detection of inflammatory cells and soluble mediators

Fig. 4

Hyaluronidase treatment is essential for an unbiased analysis of the cellular composition of synovial fluid. A, C Percentage of cells obtained after hyaluronidase treatment relative to total (before hyaluronidase + after hyaluronidase) number of cells. Cell subsets are defined as follows: neutrophils are CD15+CD16+, monocyte/macrophages are CD15-CD14+, T cells are CD3+, and B cells are CD19+. Each dot or line represents one patient. n=5–6 patients. B Percentages from A plotted against the number of live cells per mL in the total SF (before hyaluronidase + after hyaluronidase) SF determined using flow count beads. D Percentages of CD4+ and CD8+ cells present within the CD3+ cell population obtained before hyaluronidase treatment (before) or after (after). E CD69 expression on CD3+ cells. Flowcytometry plot of one representative donor is shown together with the summary of all patients. The expression of the cellular marker is indicated by ΔMFI which is defined as MFI of the antibody staining (dark gray)—MFI of the isotype control (light gray). F CD69 expression on CD4+ and CD8+ cells. G Percentage of naĂ¯ve (CD20+CD27−) and memory (CD20+CD27+) B cells, as well as plasmablast/plasma cells (CD20-CD27+) present within the CD19+ population before and after hyaluronidase are depicted. Each line is one patient. n=7–8 patients. Wilcoxon signed rank test was performed.

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