Effect of anaesthetic technique on the natural killer cell anti-tumour activity of serum from women undergoing breast cancer surgery: a pilot study  A.

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Effect of anaesthetic technique on the natural killer cell anti-tumour activity of serum from women undergoing breast cancer surgery: a pilot study  A. Buckley, S. McQuaid, P. Johnson, D.J. Buggy  British Journal of Anaesthesia  Volume 113, Pages i56-i62 (July 2014) DOI: 10.1093/bja/aeu200 Copyright © 2014 The Author(s) Terms and Conditions

Fig 1 NK receptor expression on CD56+ cells cultured in subject serum-supplemented media. The percentage of CD56+ cells expressing CD16 after 24 h culture in serum-supplemented media for normal healthy control, and pre and postoperative (post) PPA or GA study subjects was determined by surface staining and flow cytometry. (a) Mean normalized percentages of CD56+ cells expressing each receptor for 10 study serum samples and three healthy NK cell donors with standard error represented by error bars. Each experiment was carried out in triplicate. Statistical analysis was performed using the Newman–Keuls one-way anova and compared either pre- and postoperative PPA groups or pre- and postoperative GA groups (***P≤0.001). (b) Results from one replicate from one donor/serum set. British Journal of Anaesthesia 2014 113, i56-i62DOI: (10.1093/bja/aeu200) Copyright © 2014 The Author(s) Terms and Conditions

Fig 2 CD107a expression on CD56+ cells cultured in subject serum-supplemented media. The percentage of CD56+ cells expressing CD107a after 24 h culture in serum-supplemented media, either alone or with HCC tumour cells, for normal healthy control, and pre- and postoperative (post) PPA or GA study subject was determined by surface staining and flow cytometry. (a) The mean normalized percentage of CD56+ cells expressing CD107a for 10 study serum samples and three healthy NK cell donors with standard error represented by error bars. Each experiment was carried out in triplicate. Statistical analysis was performed using the Newman–Keuls one-way anova and compared NK cells cultured alone with NK cells cultured with HCC cells, for each serum type (**P≤0.01, ***P≤0.001). (b) Representative results from one replicate from one donor/serum set. British Journal of Anaesthesia 2014 113, i56-i62DOI: (10.1093/bja/aeu200) Copyright © 2014 The Author(s) Terms and Conditions

Fig 3 NK cell induced apoptosis in HCC cells when cultured in subject serum-supplemented media. The percentage of HCC cells in late apoptosis after 24 h culture in serum-supplemented media, either alone, or with CD56+ NK cells, for normal healthy control, and pre- and postoperative (post) PPA or GA study subjects was determined by dual staining with FITC-Annexin V and propidium iodide. The percentage of cells in late apoptosis was determined by flow cytometry (upper right histogram quadrant). (a) The mean normalized percentage of dead HCC cells for 10 study serum samples and three healthy NK cell donors with standard error represented by error bars. Each experiment was carried out in triplicate. Statistical analysis was performed using the Newman–Keuls one-way anova and compared HCC cells cultured alone with HCC cells cultured with CD56+ cells for each serum type (***P≤0.001). (b) Representative results from one replicate from one donor/serum sample with the upper right quadrant showing cells in late apoptosis. British Journal of Anaesthesia 2014 113, i56-i62DOI: (10.1093/bja/aeu200) Copyright © 2014 The Author(s) Terms and Conditions