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Circulating NK cells and their subsets in Beh ç et's disease

View Article: PubMed Central - PubMed

ABSTRACT

Behçet's disease (BD) is an autoinflammatory, chronic relapsing/remitting disease of unknown aetiology with both innate and acquired immune cells implicated in disease pathogenesis. Peripheral blood natural killer (NK) cells and their CD56Dim/CD56Bright subsets were surface phenotyped using CD27 and CD16 surface markers in 60 BD patients compared to 60 healthy controls (HCs). Functional potential was assessed by production of interferon (IFN)‐γ, granzyme B, perforin and the expression of degranulation marker CD107a. The effects of disease activity (BDActiveversus BDQuiet) and BD medication on NK cells were also investigated. Peripheral blood NK cells (P < 0·0001) and their constituent CD56Dim (P < 0·0001) and CD56Bright (P = 0·0015) subsets were depleted significantly in BD patients compared to HCs, and especially in those with active disease (BDActive) (P < 0·0001). BD patients taking azathioprine also had significantly depleted NK cells compared to HCs (P < 0·0001). A stepwise multivariate linear regression model confirmed BD activity and azathioprine therapy as significant independent predictor variables of peripheral blood NK percentage (P < 0·001). In general, CD56Dim cells produced more perforin (P < 0·0001) and granzyme B (P < 0·01) expressed higher CD16 levels (P < 0·0001) compared to CD56Bright cells, confirming their increased cytotoxic potential with overall higher NK cell CD107a expression in BD compared to HCs (P < 0·01). Interestingly, IFN‐γ production and CD27 expression were not significantly different between CD56Dim/CD56Bright subsets. In conclusion, both BD activity and azathioprine therapy have significant independent depletive effects on the peripheral blood NK cell compartment.

No MeSH data available.


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CD56Dim and CD56Bright natural killer (NK) subsets are phenotypically and functionally distinct. (a) Difference in percentage of CD56Dim (far left) and CD56Bright subsets (middle) as a percentage of gated lymphocytes with change in CD56Bright subset expressed as a percentage of total NK cells (far right). (b) Flow cytometry histogram plots showing intensity of surface marker expression of CD27 and CD16 and intracellular cytokine staining for interferon (IFN)‐γ, perforin and granzyme B following 5‐h phorbol myristate actate (PMA)/ionomycin stimulation (representative plots from healthy control sample are shown). (c) Summary median fluorescent intensities (MFI) of gated positive events for CD27, CD16, IFN‐γ, perforin and granzyme B showing relative differences in expression intensities of markers on each of NK subset (CD56Dim and CD56Bright) in healthy controls (HC) and in Behçet's disease (BD) patients. *P < 0·05; **P < 0·01; ***P < 0·001; n.s. = not significant.
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cei12939-fig-0002: CD56Dim and CD56Bright natural killer (NK) subsets are phenotypically and functionally distinct. (a) Difference in percentage of CD56Dim (far left) and CD56Bright subsets (middle) as a percentage of gated lymphocytes with change in CD56Bright subset expressed as a percentage of total NK cells (far right). (b) Flow cytometry histogram plots showing intensity of surface marker expression of CD27 and CD16 and intracellular cytokine staining for interferon (IFN)‐γ, perforin and granzyme B following 5‐h phorbol myristate actate (PMA)/ionomycin stimulation (representative plots from healthy control sample are shown). (c) Summary median fluorescent intensities (MFI) of gated positive events for CD27, CD16, IFN‐γ, perforin and granzyme B showing relative differences in expression intensities of markers on each of NK subset (CD56Dim and CD56Bright) in healthy controls (HC) and in Behçet's disease (BD) patients. *P < 0·05; **P < 0·01; ***P < 0·001; n.s. = not significant.

Mentions: BD patients had significantly depleted CD56Dim (HC = 9·99 ± 0·74% versus BD = 6·02 ± 0·65%) (P < 0·0001) and CD56Bright (HC = 0·62 ± 0·05% versus BD = 0·51 ± 0·06%) (P = 0·0015) NK subsets compared to healthy controls relative to total lymphocytes. Overall, this led to a net increase in the proportion of the CD56Bright subset relative to total NK cells in BD (12·80 ± 1·82%) compared to HC (7·05 ± 0·59%) (P = 0·0342) (Fig. 2a). Therefore, BD appears to be associated with a shift in NK subsets with a relative increase in the CD56Bright compared to CD56Dim cells on the background of overall peripheral blood NK depletion.


Circulating NK cells and their subsets in Beh ç et's disease
CD56Dim and CD56Bright natural killer (NK) subsets are phenotypically and functionally distinct. (a) Difference in percentage of CD56Dim (far left) and CD56Bright subsets (middle) as a percentage of gated lymphocytes with change in CD56Bright subset expressed as a percentage of total NK cells (far right). (b) Flow cytometry histogram plots showing intensity of surface marker expression of CD27 and CD16 and intracellular cytokine staining for interferon (IFN)‐γ, perforin and granzyme B following 5‐h phorbol myristate actate (PMA)/ionomycin stimulation (representative plots from healthy control sample are shown). (c) Summary median fluorescent intensities (MFI) of gated positive events for CD27, CD16, IFN‐γ, perforin and granzyme B showing relative differences in expression intensities of markers on each of NK subset (CD56Dim and CD56Bright) in healthy controls (HC) and in Behçet's disease (BD) patients. *P < 0·05; **P < 0·01; ***P < 0·001; n.s. = not significant.
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cei12939-fig-0002: CD56Dim and CD56Bright natural killer (NK) subsets are phenotypically and functionally distinct. (a) Difference in percentage of CD56Dim (far left) and CD56Bright subsets (middle) as a percentage of gated lymphocytes with change in CD56Bright subset expressed as a percentage of total NK cells (far right). (b) Flow cytometry histogram plots showing intensity of surface marker expression of CD27 and CD16 and intracellular cytokine staining for interferon (IFN)‐γ, perforin and granzyme B following 5‐h phorbol myristate actate (PMA)/ionomycin stimulation (representative plots from healthy control sample are shown). (c) Summary median fluorescent intensities (MFI) of gated positive events for CD27, CD16, IFN‐γ, perforin and granzyme B showing relative differences in expression intensities of markers on each of NK subset (CD56Dim and CD56Bright) in healthy controls (HC) and in Behçet's disease (BD) patients. *P < 0·05; **P < 0·01; ***P < 0·001; n.s. = not significant.
Mentions: BD patients had significantly depleted CD56Dim (HC = 9·99 ± 0·74% versus BD = 6·02 ± 0·65%) (P < 0·0001) and CD56Bright (HC = 0·62 ± 0·05% versus BD = 0·51 ± 0·06%) (P = 0·0015) NK subsets compared to healthy controls relative to total lymphocytes. Overall, this led to a net increase in the proportion of the CD56Bright subset relative to total NK cells in BD (12·80 ± 1·82%) compared to HC (7·05 ± 0·59%) (P = 0·0342) (Fig. 2a). Therefore, BD appears to be associated with a shift in NK subsets with a relative increase in the CD56Bright compared to CD56Dim cells on the background of overall peripheral blood NK depletion.

View Article: PubMed Central - PubMed

ABSTRACT

Beh&ccedil;et's disease (BD) is an autoinflammatory, chronic relapsing/remitting disease of unknown aetiology with both innate and acquired immune cells implicated in disease pathogenesis. Peripheral blood natural killer (NK) cells and their CD56Dim/CD56Bright subsets were surface phenotyped using CD27 and CD16 surface markers in 60 BD patients compared to 60 healthy controls (HCs). Functional potential was assessed by production of interferon (IFN)&#8208;&gamma;, granzyme B, perforin and the expression of degranulation marker CD107a. The effects of disease activity (BDActiveversus BDQuiet) and BD medication on NK cells were also investigated. Peripheral blood NK cells (P&thinsp;&lt;&thinsp;0&middot;0001) and their constituent CD56Dim (P&thinsp;&lt;&thinsp;0&middot;0001) and CD56Bright (P&thinsp;=&thinsp;0&middot;0015) subsets were depleted significantly in BD patients compared to HCs, and especially in those with active disease (BDActive) (P&thinsp;&lt;&thinsp;0&middot;0001). BD patients taking azathioprine also had significantly depleted NK cells compared to HCs (P&thinsp;&lt;&thinsp;0&middot;0001). A stepwise multivariate linear regression model confirmed BD activity and azathioprine therapy as significant independent predictor variables of peripheral blood NK percentage (P&thinsp;&lt;&thinsp;0&middot;001). In general, CD56Dim cells produced more perforin (P&thinsp;&lt;&thinsp;0&middot;0001) and granzyme B (P&thinsp;&lt;&thinsp;0&middot;01) expressed higher CD16 levels (P&thinsp;&lt;&thinsp;0&middot;0001) compared to CD56Bright cells, confirming their increased cytotoxic potential with overall higher NK cell CD107a expression in BD compared to HCs (P&thinsp;&lt;&thinsp;0&middot;01). Interestingly, IFN&#8208;&gamma; production and CD27 expression were not significantly different between CD56Dim/CD56Bright subsets. In conclusion, both BD activity and azathioprine therapy have significant independent depletive effects on the peripheral blood NK cell compartment.

No MeSH data available.


Related in: MedlinePlus