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Fasting enhances TRAIL-mediated liver natural killer cell activity via HSP70 upregulation.

Dang VT, Tanabe K, Tanaka Y, Tokumoto N, Misumi T, Saeki Y, Fujikuni N, Ohdan H - PLoS ONE (2014)

Bottom Line: Liver natural killer cells also showed high TRAIL-mediated antitumor function in response to 3-day fasting.Treatment of liver lymphocytes with 50 µg/mL of recombinant heat shock protein 70 led to the upregulation of both TRAIL and CD69 in liver natural killer cells (n = 6, p <0.05).These findings indicate that acute fasting enhances TRAIL-mediated liver natural killer cell activity against neoplastic cells through upregulation of heat shock protein 70.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterological and Transplant Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

ABSTRACT
Acute starvation, which is frequently observed in clinical practice, sometimes augments the cytolytic activity of natural killer cells against neoplastic cells. In this study, we investigated the molecular mechanisms underlying the enhancement of natural killer cell function by fasting in mice. The total number of liver resident natural killer cells in a unit weight of liver tissue obtained from C57BL/6J mice did not change after a 3-day fast, while the proportions of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)+ and CD69+ natural killer cells were significantly elevated (n = 7, p <0.01), as determined by flow cytometric analysis. Furthermore, we found that TRAIL- natural killer cells that were adoptively transferred into Rag-2-/- γ chain-/- mice could convert into TRAIL+ natural killer cells in fasted mice at a higher proportion than in fed mice. Liver natural killer cells also showed high TRAIL-mediated antitumor function in response to 3-day fasting. Since these fasted mice highly expressed heat shock protein 70 (n = 7, p <0.05) in liver tissues, as determined by western blot, the role of this protein in natural killer cell activation was investigated. Treatment of liver lymphocytes with 50 µg/mL of recombinant heat shock protein 70 led to the upregulation of both TRAIL and CD69 in liver natural killer cells (n = 6, p <0.05). In addition, HSP70 neutralization by intraperitoneally injecting an anti- heat shock protein 70 monoclonal antibody into mice prior to fasting led to the downregulation of TRAIL expression (n = 6, p <0.05). These findings indicate that acute fasting enhances TRAIL-mediated liver natural killer cell activity against neoplastic cells through upregulation of heat shock protein 70.

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Analysis of functional markers in liver natural killer cells and their CD49a+ DX5− and CD49a− DX5+ subgroups.Liver lymphocytes from fed and 3-day-fasted mice were simultaneously stained with monoclonal antibodies against DX5, CD49a, TRAIL, CD69, NKG2D, NKp46, and FasL. (A) Representative dot plots of gated TCRβ− NK1.1+ natural killer (NK) cells and its two subsets, CD49a+ DX5− and CD49a− DX5+ NK cells, in fed and fasted mice are presented. Histograms show the expression of TRAIL, CD69, NKG2D, NKp46, and FasL (solid lines) on whole NK cells and their subsets with the percentage of NK cells that are positive for those markers, dotted lines present negative control. (B) Bar graph shows the mean percentage plus standard deviation of NK cells that are positive for TRAIL, CD69, NKG2D, NKp46, or FasL. (C) The proportion of NK cell subsets, CD49a+ DX5− and CD49a− DX5+ NK cells, in fed and fasted mice are shown as mean ratio plus standard deviation; *p <0.05 as analyzed by the independent samples T test.
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pone-0110748-g003: Analysis of functional markers in liver natural killer cells and their CD49a+ DX5− and CD49a− DX5+ subgroups.Liver lymphocytes from fed and 3-day-fasted mice were simultaneously stained with monoclonal antibodies against DX5, CD49a, TRAIL, CD69, NKG2D, NKp46, and FasL. (A) Representative dot plots of gated TCRβ− NK1.1+ natural killer (NK) cells and its two subsets, CD49a+ DX5− and CD49a− DX5+ NK cells, in fed and fasted mice are presented. Histograms show the expression of TRAIL, CD69, NKG2D, NKp46, and FasL (solid lines) on whole NK cells and their subsets with the percentage of NK cells that are positive for those markers, dotted lines present negative control. (B) Bar graph shows the mean percentage plus standard deviation of NK cells that are positive for TRAIL, CD69, NKG2D, NKp46, or FasL. (C) The proportion of NK cell subsets, CD49a+ DX5− and CD49a− DX5+ NK cells, in fed and fasted mice are shown as mean ratio plus standard deviation; *p <0.05 as analyzed by the independent samples T test.

Mentions: Analysis of other functional markers of NK cells indicated that whole liver NK cells from 3-day fasted mice highly expressed not only TRAIL and CD69 but also NKp46 when compared with fed mice. There was no significant difference in NKG2D or FasL expression (Figure 3A, B). Additionally, changes in CD49a and DX5 phenotype characteristics in NK cells were examined based on a report that recently demonstrated that liver-resident CD3− NK1.1+ NK subsets are characterized according to the differential expression of CD49a and DX5 [24]. While proportions of CD49a− DX5+ NK cells significantly decreased in fasted mice, the proportion of CD49a+ DX5− NK cells, which highly expressed TRAIL and CD69, significantly increased (Figure 3A, C). Taken together, our results indicate that, under 3-day fasting, TRAIL and CD69 are highly expressed in mouse liver-resident CD49a+ DX5− NK cells.


Fasting enhances TRAIL-mediated liver natural killer cell activity via HSP70 upregulation.

Dang VT, Tanabe K, Tanaka Y, Tokumoto N, Misumi T, Saeki Y, Fujikuni N, Ohdan H - PLoS ONE (2014)

Analysis of functional markers in liver natural killer cells and their CD49a+ DX5− and CD49a− DX5+ subgroups.Liver lymphocytes from fed and 3-day-fasted mice were simultaneously stained with monoclonal antibodies against DX5, CD49a, TRAIL, CD69, NKG2D, NKp46, and FasL. (A) Representative dot plots of gated TCRβ− NK1.1+ natural killer (NK) cells and its two subsets, CD49a+ DX5− and CD49a− DX5+ NK cells, in fed and fasted mice are presented. Histograms show the expression of TRAIL, CD69, NKG2D, NKp46, and FasL (solid lines) on whole NK cells and their subsets with the percentage of NK cells that are positive for those markers, dotted lines present negative control. (B) Bar graph shows the mean percentage plus standard deviation of NK cells that are positive for TRAIL, CD69, NKG2D, NKp46, or FasL. (C) The proportion of NK cell subsets, CD49a+ DX5− and CD49a− DX5+ NK cells, in fed and fasted mice are shown as mean ratio plus standard deviation; *p <0.05 as analyzed by the independent samples T test.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4214715&req=5

pone-0110748-g003: Analysis of functional markers in liver natural killer cells and their CD49a+ DX5− and CD49a− DX5+ subgroups.Liver lymphocytes from fed and 3-day-fasted mice were simultaneously stained with monoclonal antibodies against DX5, CD49a, TRAIL, CD69, NKG2D, NKp46, and FasL. (A) Representative dot plots of gated TCRβ− NK1.1+ natural killer (NK) cells and its two subsets, CD49a+ DX5− and CD49a− DX5+ NK cells, in fed and fasted mice are presented. Histograms show the expression of TRAIL, CD69, NKG2D, NKp46, and FasL (solid lines) on whole NK cells and their subsets with the percentage of NK cells that are positive for those markers, dotted lines present negative control. (B) Bar graph shows the mean percentage plus standard deviation of NK cells that are positive for TRAIL, CD69, NKG2D, NKp46, or FasL. (C) The proportion of NK cell subsets, CD49a+ DX5− and CD49a− DX5+ NK cells, in fed and fasted mice are shown as mean ratio plus standard deviation; *p <0.05 as analyzed by the independent samples T test.
Mentions: Analysis of other functional markers of NK cells indicated that whole liver NK cells from 3-day fasted mice highly expressed not only TRAIL and CD69 but also NKp46 when compared with fed mice. There was no significant difference in NKG2D or FasL expression (Figure 3A, B). Additionally, changes in CD49a and DX5 phenotype characteristics in NK cells were examined based on a report that recently demonstrated that liver-resident CD3− NK1.1+ NK subsets are characterized according to the differential expression of CD49a and DX5 [24]. While proportions of CD49a− DX5+ NK cells significantly decreased in fasted mice, the proportion of CD49a+ DX5− NK cells, which highly expressed TRAIL and CD69, significantly increased (Figure 3A, C). Taken together, our results indicate that, under 3-day fasting, TRAIL and CD69 are highly expressed in mouse liver-resident CD49a+ DX5− NK cells.

Bottom Line: Liver natural killer cells also showed high TRAIL-mediated antitumor function in response to 3-day fasting.Treatment of liver lymphocytes with 50 µg/mL of recombinant heat shock protein 70 led to the upregulation of both TRAIL and CD69 in liver natural killer cells (n = 6, p <0.05).These findings indicate that acute fasting enhances TRAIL-mediated liver natural killer cell activity against neoplastic cells through upregulation of heat shock protein 70.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterological and Transplant Surgery, Applied Life Sciences, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

ABSTRACT
Acute starvation, which is frequently observed in clinical practice, sometimes augments the cytolytic activity of natural killer cells against neoplastic cells. In this study, we investigated the molecular mechanisms underlying the enhancement of natural killer cell function by fasting in mice. The total number of liver resident natural killer cells in a unit weight of liver tissue obtained from C57BL/6J mice did not change after a 3-day fast, while the proportions of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)+ and CD69+ natural killer cells were significantly elevated (n = 7, p <0.01), as determined by flow cytometric analysis. Furthermore, we found that TRAIL- natural killer cells that were adoptively transferred into Rag-2-/- γ chain-/- mice could convert into TRAIL+ natural killer cells in fasted mice at a higher proportion than in fed mice. Liver natural killer cells also showed high TRAIL-mediated antitumor function in response to 3-day fasting. Since these fasted mice highly expressed heat shock protein 70 (n = 7, p <0.05) in liver tissues, as determined by western blot, the role of this protein in natural killer cell activation was investigated. Treatment of liver lymphocytes with 50 µg/mL of recombinant heat shock protein 70 led to the upregulation of both TRAIL and CD69 in liver natural killer cells (n = 6, p <0.05). In addition, HSP70 neutralization by intraperitoneally injecting an anti- heat shock protein 70 monoclonal antibody into mice prior to fasting led to the downregulation of TRAIL expression (n = 6, p <0.05). These findings indicate that acute fasting enhances TRAIL-mediated liver natural killer cell activity against neoplastic cells through upregulation of heat shock protein 70.

Show MeSH
Related in: MedlinePlus