Korean J Pain.  2018 Jan;31(1):43-49. 10.3344/kjp.2018.31.1.43.

Cytotoxic activity and subset populations of peripheral blood natural killer cells in patients with chronic pain

• Yoon JJ
• Song JA
• Park SY
• Choi JI

• KMID: 2400907
• DOI: http://doi.org/10.3344/kjp.2018.31.1.43

Abstract

BACKGROUND
Chronic pain reportedly exerts complex effects on immune function. Natural killer (NK) cells are lymphocytes that play a critical role in cellular and innate immunity. This study examined changes in the subset populations and cytotoxic activity of peripheral blood NK cells in patients with chronic pain.
METHODS
Thirty patients with chronic moderate-to-severe pain (group P) and age-matched pain-free subjects (group NoP) were enrolled. Peripheral whole blood was analyzed for the percentage and expression of NK cell surface markers (CD56 and CD16) by flow cytometry. Cytotoxic activity was assayed by evaluating CD69 expression on CD3−/CD56+NK cells.
RESULTS
The percentage of NK cells among total lymphocytes was not significantly different between groups P and NoP (16.3 ± 9.3 vs. 20.2 ± 10.5%). Likewise, the percentages of two major NK cell subsets, CD56bright and CD56dim, were also not significantly different between the two groups. However, the percentage of CD56bright/CD16+ subset, was slightly but significantly increased in group P (1.0 ± 0.9%; P < 0.01) compared with group NoP (0.5 ± 0.6%). The cytotoxicity of NK cells was not different between the two groups, showing similar CD69 expression (P vs. NoP = 29.2 ± 15.2 vs. 32.0 ± 15.0%). These findings were not influenced by pain intensity, opioid use, or disease causing pain in group P.
CONCLUSIONS
NK cell cytotoxic activity and major subset populations, with the exception of an increased percentage of the CD56bright/CD16+ subset, are not significantly altered in patients with chronic severe pain.

Keyword

CD56(bright)/CD16⁺; CD69; Chronic pain; Cytotoxicity; Natural killer cell; Peripheral blood

MeSH Terms

Chronic Pain*
Flow Cytometry
Humans
Immunity, Innate
Killer Cells, Natural*
Lymphocytes

Figure

• 

  Fig. 1 Gating techniques used to identify NK subsets in whole blood samples. Each dot indicates a single cell analyzed by flow cytometry. As each cell is forced to pass through the laser beam, the cell is interrogated by the laser and scatters the light. Several detectors measure the light scattered from cells as either forward scatter (FS) or side scatter (SS); these measures are correlated with the size and granularity, respectively. The y-axis in panel (A) represents the amount of lights detected as SS, which is expressed using a linear scale. The intensity of the fluorescence obtained from cells stained with fluorescent antibodies is expressed using a logarithmic scale on the x- or y-axis of panel (A)–(C). After the lymphocytes were gated (A), NK cell subsets (CD3−/CD56+) were sequentially gated on the basis of the surface expression of CD3 (T-lymphocyte) and CD19 (B-lymphocyte)/CD14 (monocyte) (B). Finally, CD3−/CD56+ cells were gated into 2 main subsets and 5 subsets depending on the expression of CD56 and CD16 (C). The numbers in panel (C) represent the following subsets: 1) CD56bright/CD16−, 2) CD56bright/CD16+, 3) CD56dim/CD16−, 4) CD56dim/CD16+, and 5) CD56−/CD16+.

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  Fig. 2 Comparison of the percentages of 5 NK cell subsets in group NoP and P. The y-axes indicates the percentage of each NK cell subset, and each dot represents each value (percentage) of NK subsets in group NoP (•) and P (▴).

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  Fig. 3 Pain intensity (numerical rating scale, NRS) was not correlated with the increase in CD56bright/CD16+ cells (A) or percentage of cells expressing CD69 (B) ingroup P. Spearman's correlation test was used; r represents Spearman's coefficient.

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