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Effect of surgery on pancreatic tumor-dependent lymphocyte asset: modulation of natural killer cell frequency and cytotoxic function.

Iannone F, Porzia A, Peruzzi G, Birarelli P, Milana B, Sacco L, Dinatale G, Peparini N, Prezioso G, Battella S, Caronna R, Morrone S, Palmieri G, Mainiero F, Chirletti P - Pancreas (2015)

Bottom Line: Tumor burden and invasiveness establish a microenvironment that surgery could alter.This study shows a comprehensive analysis of size, dynamics, and function of peripheral lymphocyte subsets in pancreatic cancer patients before and at different times after duodenopancreatectomy.The NK cell functional ability is altered in presurgery patients; duodenopancreatectomy is associated with short-term impairment of NK function and with a long-term NK cell augmentation and reversion of the aberrant NK behavior, which may impact on immunosurveillance against residual cancer.

View Article: PubMed Central - PubMed

Affiliation: From the *Section of Interdisciplinary Surgery F. Durante, Department of Surgical Sciences, †Department of Molecular Medicine, ‡Istituto Italiano di Tecnologia, CLNS, and §Department of Experimental Medicine, Sapienza University of Rome, Italy.

ABSTRACT

Objectives: Tumor burden and invasiveness establish a microenvironment that surgery could alter. This study shows a comprehensive analysis of size, dynamics, and function of peripheral lymphocyte subsets in pancreatic cancer patients before and at different times after duodenopancreatectomy.

Methods: Lymphocyte frequency and natural cytotoxicity were evaluated by flow cytometry and in vitro assay on peripheral blood from initial and advanced-stage pancreatic cancer patients before (BS), at day 7 (PS7), and at day 30 (PS30) after surgery.

Results: An increase in natural killer (NK) cells and the diminution of B-cells occurred at PS30, whereas cytotoxicity decreased at PS7. The positive correlation between NK frequency and cytotoxicity at BS and PS7 revealed an altered NK behavior. The elevation of NK cell frequency at PS30, an initial defect in CD56bright NK, and the aberrant correlation between NK frequency and cytotoxicity remained significant in advanced-stage patients, whereas the diminution of NK cytotoxicity only affected initial stage patients.

Conclusions: The NK cell functional ability is altered in presurgery patients; duodenopancreatectomy is associated with short-term impairment of NK function and with a long-term NK cell augmentation and reversion of the aberrant NK behavior, which may impact on immunosurveillance against residual cancer.

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Modulation of NK cell cytotoxicity and dynamics of its relationship with NK cell frequency in pancreatic cancer patients undergoing DP. A, Percentage of NK cytotoxic activity in PB of pancreatic cancer patients before surgery (BS), at day 7 (PS7), and at day 30 (PS30) postsurgery, as well as age-matched controls. Bars show median and interquartile range. B, Scatter plots representing the percentage of PBMC cytotoxicity with respect to NK cell frequency; NK cells before surgery (BS, black symbols and line), at day 7 (PS7, gray symbols and line), and at day 30 (PS30, white symbols) postsurgery, as well as age-matched controls (x symbols). Lines represent the linear regression for any given pair of variables. *P < 0.05, **P < 0.005.
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Figure 2: Modulation of NK cell cytotoxicity and dynamics of its relationship with NK cell frequency in pancreatic cancer patients undergoing DP. A, Percentage of NK cytotoxic activity in PB of pancreatic cancer patients before surgery (BS), at day 7 (PS7), and at day 30 (PS30) postsurgery, as well as age-matched controls. Bars show median and interquartile range. B, Scatter plots representing the percentage of PBMC cytotoxicity with respect to NK cell frequency; NK cells before surgery (BS, black symbols and line), at day 7 (PS7, gray symbols and line), and at day 30 (PS30, white symbols) postsurgery, as well as age-matched controls (x symbols). Lines represent the linear regression for any given pair of variables. *P < 0.05, **P < 0.005.

Mentions: Natural killer cells represent an important effector of antitumor immunosurveillance, by exerting a direct cytotoxicity against tumor cells. Their increase in PB samples of pancreatic cancer patients at 30 days postsurgery prompted us to investigate NK cell cytolytic function. The NK cell’s natural cytotoxicity depends on activating receptors that interact with “stress-induced” class I-like molecules that are frequently up-regulated on malignant cells, such as in pancreatic cancer.38,39 Natural killer cell cytotoxic activity in PBMC of presurgery patients was comparable to that of controls, evaluated as the percentage of in vitro killing of a reference tumor cell line (K562) (Fig. 2A); a transient diminution occurred at day 7 postintervention and was reconstituted at 30 days after surgery (Fig. 2A); comparable results have been obtained when NK cytotoxic activity was expressed in lytic units (data not shown). Thus, the elevation of NK cell frequency at day 30 after surgery does not quantitatively correlate with the transient diminution of NK cell cytotoxicity observed at an earlier (day 7 postsurgery) time point. We thus analyzed more in depth the dynamic relationship between the abundance of PB NK cells and the in vitro cytotoxic activity in pancreatic tumor patients; at presurgery, the frequency of NK cells and the extent of PBMC cytotoxic activity were positively correlated in each individual. This relationship was maintained at day 7 time point, whereas it was absent at day 30 postintervention (Fig. 2B). Interestingly, NK cell frequency and PBMC cytotoxic activity showed no significant correlation in control subjects; this evidence has been previously reported in the literature and explained by the large variability in individual immune cell state of activation.40,41 Collectively taken, these data suggest that although present in a normal percentage, the lytic behavior of presurgery patients’ NK cells is altered; moreover, their cytotoxic activity is reduced at a short time point after surgery. Later on after intervention (30 days), the elevation of frequency, the recovery of NK cytotoxic activity, and the loss of the correlation between NK cell frequency and cytotoxicity collectively suggest a normalization of NK cell behavior.


Effect of surgery on pancreatic tumor-dependent lymphocyte asset: modulation of natural killer cell frequency and cytotoxic function.

Iannone F, Porzia A, Peruzzi G, Birarelli P, Milana B, Sacco L, Dinatale G, Peparini N, Prezioso G, Battella S, Caronna R, Morrone S, Palmieri G, Mainiero F, Chirletti P - Pancreas (2015)

Modulation of NK cell cytotoxicity and dynamics of its relationship with NK cell frequency in pancreatic cancer patients undergoing DP. A, Percentage of NK cytotoxic activity in PB of pancreatic cancer patients before surgery (BS), at day 7 (PS7), and at day 30 (PS30) postsurgery, as well as age-matched controls. Bars show median and interquartile range. B, Scatter plots representing the percentage of PBMC cytotoxicity with respect to NK cell frequency; NK cells before surgery (BS, black symbols and line), at day 7 (PS7, gray symbols and line), and at day 30 (PS30, white symbols) postsurgery, as well as age-matched controls (x symbols). Lines represent the linear regression for any given pair of variables. *P < 0.05, **P < 0.005.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4358707&req=5

Figure 2: Modulation of NK cell cytotoxicity and dynamics of its relationship with NK cell frequency in pancreatic cancer patients undergoing DP. A, Percentage of NK cytotoxic activity in PB of pancreatic cancer patients before surgery (BS), at day 7 (PS7), and at day 30 (PS30) postsurgery, as well as age-matched controls. Bars show median and interquartile range. B, Scatter plots representing the percentage of PBMC cytotoxicity with respect to NK cell frequency; NK cells before surgery (BS, black symbols and line), at day 7 (PS7, gray symbols and line), and at day 30 (PS30, white symbols) postsurgery, as well as age-matched controls (x symbols). Lines represent the linear regression for any given pair of variables. *P < 0.05, **P < 0.005.
Mentions: Natural killer cells represent an important effector of antitumor immunosurveillance, by exerting a direct cytotoxicity against tumor cells. Their increase in PB samples of pancreatic cancer patients at 30 days postsurgery prompted us to investigate NK cell cytolytic function. The NK cell’s natural cytotoxicity depends on activating receptors that interact with “stress-induced” class I-like molecules that are frequently up-regulated on malignant cells, such as in pancreatic cancer.38,39 Natural killer cell cytotoxic activity in PBMC of presurgery patients was comparable to that of controls, evaluated as the percentage of in vitro killing of a reference tumor cell line (K562) (Fig. 2A); a transient diminution occurred at day 7 postintervention and was reconstituted at 30 days after surgery (Fig. 2A); comparable results have been obtained when NK cytotoxic activity was expressed in lytic units (data not shown). Thus, the elevation of NK cell frequency at day 30 after surgery does not quantitatively correlate with the transient diminution of NK cell cytotoxicity observed at an earlier (day 7 postsurgery) time point. We thus analyzed more in depth the dynamic relationship between the abundance of PB NK cells and the in vitro cytotoxic activity in pancreatic tumor patients; at presurgery, the frequency of NK cells and the extent of PBMC cytotoxic activity were positively correlated in each individual. This relationship was maintained at day 7 time point, whereas it was absent at day 30 postintervention (Fig. 2B). Interestingly, NK cell frequency and PBMC cytotoxic activity showed no significant correlation in control subjects; this evidence has been previously reported in the literature and explained by the large variability in individual immune cell state of activation.40,41 Collectively taken, these data suggest that although present in a normal percentage, the lytic behavior of presurgery patients’ NK cells is altered; moreover, their cytotoxic activity is reduced at a short time point after surgery. Later on after intervention (30 days), the elevation of frequency, the recovery of NK cytotoxic activity, and the loss of the correlation between NK cell frequency and cytotoxicity collectively suggest a normalization of NK cell behavior.

Bottom Line: Tumor burden and invasiveness establish a microenvironment that surgery could alter.This study shows a comprehensive analysis of size, dynamics, and function of peripheral lymphocyte subsets in pancreatic cancer patients before and at different times after duodenopancreatectomy.The NK cell functional ability is altered in presurgery patients; duodenopancreatectomy is associated with short-term impairment of NK function and with a long-term NK cell augmentation and reversion of the aberrant NK behavior, which may impact on immunosurveillance against residual cancer.

View Article: PubMed Central - PubMed

Affiliation: From the *Section of Interdisciplinary Surgery F. Durante, Department of Surgical Sciences, †Department of Molecular Medicine, ‡Istituto Italiano di Tecnologia, CLNS, and §Department of Experimental Medicine, Sapienza University of Rome, Italy.

ABSTRACT

Objectives: Tumor burden and invasiveness establish a microenvironment that surgery could alter. This study shows a comprehensive analysis of size, dynamics, and function of peripheral lymphocyte subsets in pancreatic cancer patients before and at different times after duodenopancreatectomy.

Methods: Lymphocyte frequency and natural cytotoxicity were evaluated by flow cytometry and in vitro assay on peripheral blood from initial and advanced-stage pancreatic cancer patients before (BS), at day 7 (PS7), and at day 30 (PS30) after surgery.

Results: An increase in natural killer (NK) cells and the diminution of B-cells occurred at PS30, whereas cytotoxicity decreased at PS7. The positive correlation between NK frequency and cytotoxicity at BS and PS7 revealed an altered NK behavior. The elevation of NK cell frequency at PS30, an initial defect in CD56bright NK, and the aberrant correlation between NK frequency and cytotoxicity remained significant in advanced-stage patients, whereas the diminution of NK cytotoxicity only affected initial stage patients.

Conclusions: The NK cell functional ability is altered in presurgery patients; duodenopancreatectomy is associated with short-term impairment of NK function and with a long-term NK cell augmentation and reversion of the aberrant NK behavior, which may impact on immunosurveillance against residual cancer.

Show MeSH
Related in: MedlinePlus