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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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The effect of recombinant heat shock protein 70 on natural killer cell proliferation and TRAIL and CD69 expression.Isolated liver lymphocytes (2 million cells/well) were cultured with mouse recombinant heat shock protein (HSP) 70 at various concentrations: 0, 0.5, 5, or 50 µg/mL. After 3 days of culture, the lymphocytes were harvested for phenotypic determination. (A) Representative flow cytometric analysis of TRAIL and CD69 expression in TCRβ− NK1.1+ natural killer (NK) cells is shown. The dotted lines represent the expression distribution in the negative control cells and the solid lines (shaded areas) with numbers indicate the distribution of TRAIL+ and CD69+ NK cells. (B) TCRβ− NK1.1+ NK cell number per well, (C) TRAIL+ and (D) CD69+ NK cell percentages are shown in bar graphs as mean plus standard deviation (n = 6). Data were statistically analyzed using the independent samples T test; *p <0.05.
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pone-0110748-g007: The effect of recombinant heat shock protein 70 on natural killer cell proliferation and TRAIL and CD69 expression.Isolated liver lymphocytes (2 million cells/well) were cultured with mouse recombinant heat shock protein (HSP) 70 at various concentrations: 0, 0.5, 5, or 50 µg/mL. After 3 days of culture, the lymphocytes were harvested for phenotypic determination. (A) Representative flow cytometric analysis of TRAIL and CD69 expression in TCRβ− NK1.1+ natural killer (NK) cells is shown. The dotted lines represent the expression distribution in the negative control cells and the solid lines (shaded areas) with numbers indicate the distribution of TRAIL+ and CD69+ NK cells. (B) TCRβ− NK1.1+ NK cell number per well, (C) TRAIL+ and (D) CD69+ NK cell percentages are shown in bar graphs as mean plus standard deviation (n = 6). Data were statistically analyzed using the independent samples T test; *p <0.05.

Mentions: The contribution of HSP70 to NK cell activation was assessed in vitro by examining the phenotypic characteristics of mouse liver NK cells after culturing liver lymphocytes with IL-2 and different concentrations of recombinant HSP70 (0 µg/mL as the control, 0.5, 5, or 50 µg/mL) for 3 days. Treatment with ≥5 µg/mL HSP70 induced NK cell proliferation (p <0.05; Figure 7B), whereas treatment with 50 µg/mL HSP70 led to an upregulation of TRAIL and CD69 expression in liver NK cells as compared to the control (Figure 7C, D).


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)

The effect of recombinant heat shock protein 70 on natural killer cell proliferation and TRAIL and CD69 expression.Isolated liver lymphocytes (2 million cells/well) were cultured with mouse recombinant heat shock protein (HSP) 70 at various concentrations: 0, 0.5, 5, or 50 µg/mL. After 3 days of culture, the lymphocytes were harvested for phenotypic determination. (A) Representative flow cytometric analysis of TRAIL and CD69 expression in TCRβ− NK1.1+ natural killer (NK) cells is shown. The dotted lines represent the expression distribution in the negative control cells and the solid lines (shaded areas) with numbers indicate the distribution of TRAIL+ and CD69+ NK cells. (B) TCRβ− NK1.1+ NK cell number per well, (C) TRAIL+ and (D) CD69+ NK cell percentages are shown in bar graphs as mean plus standard deviation (n = 6). Data were statistically analyzed using the independent samples T test; *p <0.05.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0110748-g007: The effect of recombinant heat shock protein 70 on natural killer cell proliferation and TRAIL and CD69 expression.Isolated liver lymphocytes (2 million cells/well) were cultured with mouse recombinant heat shock protein (HSP) 70 at various concentrations: 0, 0.5, 5, or 50 µg/mL. After 3 days of culture, the lymphocytes were harvested for phenotypic determination. (A) Representative flow cytometric analysis of TRAIL and CD69 expression in TCRβ− NK1.1+ natural killer (NK) cells is shown. The dotted lines represent the expression distribution in the negative control cells and the solid lines (shaded areas) with numbers indicate the distribution of TRAIL+ and CD69+ NK cells. (B) TCRβ− NK1.1+ NK cell number per well, (C) TRAIL+ and (D) CD69+ NK cell percentages are shown in bar graphs as mean plus standard deviation (n = 6). Data were statistically analyzed using the independent samples T test; *p <0.05.
Mentions: The contribution of HSP70 to NK cell activation was assessed in vitro by examining the phenotypic characteristics of mouse liver NK cells after culturing liver lymphocytes with IL-2 and different concentrations of recombinant HSP70 (0 µg/mL as the control, 0.5, 5, or 50 µg/mL) for 3 days. Treatment with ≥5 µg/mL HSP70 induced NK cell proliferation (p <0.05; Figure 7B), whereas treatment with 50 µg/mL HSP70 led to an upregulation of TRAIL and CD69 expression in liver NK cells as compared to the control (Figure 7C, D).

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