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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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Phenotype of liver natural killer cells under starvation.Isolated liver lymphocytes from 3 mouse groups were stained with monoclonal antibodies against the cell surface markers TRAIL, CD69, CD122, and CD25 prior to analysis by flow cytometry. Representative natural killer (NK) cell phenotype analyses from (A) fed, (B) 1-day-fasted and (C) 3-day-fasted mice are presented in dot plots and histograms. TCRβ− NK1.1+ cells were gated as NK cells. The dotted lines represent the negative control. The distribution of TRAIL, CD69, CD122, and CD25 expression in NK cells is indicated by the solid lines (shaded areas) and the percentage and mean fluorescence intensity (MFI) of positive cells are provided. (D) The percentages or (E) MFI of liver NK cells that are positive for TRAIL, CD69, CD122, and CD25 are shown in bar graphs as mean plus standard deviation. (F) Dot plots of representative data and (G) bar graph present the mean plus standard deviation of proportion of NK cell subsets regarding to TRAIL and CD69 expression in fed and fasted mice; *p <0.05, **p <0.01 as analyzed by the independent samples T test.
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pone-0110748-g001: Phenotype of liver natural killer cells under starvation.Isolated liver lymphocytes from 3 mouse groups were stained with monoclonal antibodies against the cell surface markers TRAIL, CD69, CD122, and CD25 prior to analysis by flow cytometry. Representative natural killer (NK) cell phenotype analyses from (A) fed, (B) 1-day-fasted and (C) 3-day-fasted mice are presented in dot plots and histograms. TCRβ− NK1.1+ cells were gated as NK cells. The dotted lines represent the negative control. The distribution of TRAIL, CD69, CD122, and CD25 expression in NK cells is indicated by the solid lines (shaded areas) and the percentage and mean fluorescence intensity (MFI) of positive cells are provided. (D) The percentages or (E) MFI of liver NK cells that are positive for TRAIL, CD69, CD122, and CD25 are shown in bar graphs as mean plus standard deviation. (F) Dot plots of representative data and (G) bar graph present the mean plus standard deviation of proportion of NK cell subsets regarding to TRAIL and CD69 expression in fed and fasted mice; *p <0.05, **p <0.01 as analyzed by the independent samples T test.

Mentions: Electronically gated TCRβ− NK1.1+ NK cells and NK cell markers from a representative fed, 1-day-fasted, or 3-day-fasted mouse are shown in Figure 1 A–C. Notably, compared to fed mice, 3-day-fasted mice showed significantly higher proportions of TRAIL and CD69 in liver NK cells (Figure 1D). Mean fluorescence intensity (MFI) of TRAIL or CD69 positive NK cells showed no significant differences among the groups (Figure 1E). Next, the distribution analysis of CD69 and TRAIL expression revealed that the proportion of CD69+TRAIL+ double positive NK cells significantly increased in fasted mice, while CD69− TRAIL− NK cells significantly decreased. The proportion of CD69+ TRAIL− cells also increased (Figure 1F, G). There was no difference in CD122 and CD25 expression in liver NK cells among the groups (Figure 1D, E) as well as in splenic NK cells (data not shown). The proportion of NK cells in the liver mononuclear cell fractions from 3-day-fasted mice did not differ from that from fed mice (Figure S1A).


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)

Phenotype of liver natural killer cells under starvation.Isolated liver lymphocytes from 3 mouse groups were stained with monoclonal antibodies against the cell surface markers TRAIL, CD69, CD122, and CD25 prior to analysis by flow cytometry. Representative natural killer (NK) cell phenotype analyses from (A) fed, (B) 1-day-fasted and (C) 3-day-fasted mice are presented in dot plots and histograms. TCRβ− NK1.1+ cells were gated as NK cells. The dotted lines represent the negative control. The distribution of TRAIL, CD69, CD122, and CD25 expression in NK cells is indicated by the solid lines (shaded areas) and the percentage and mean fluorescence intensity (MFI) of positive cells are provided. (D) The percentages or (E) MFI of liver NK cells that are positive for TRAIL, CD69, CD122, and CD25 are shown in bar graphs as mean plus standard deviation. (F) Dot plots of representative data and (G) bar graph present the mean plus standard deviation of proportion of NK cell subsets regarding to TRAIL and CD69 expression in fed and fasted mice; *p <0.05, **p <0.01 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-g001: Phenotype of liver natural killer cells under starvation.Isolated liver lymphocytes from 3 mouse groups were stained with monoclonal antibodies against the cell surface markers TRAIL, CD69, CD122, and CD25 prior to analysis by flow cytometry. Representative natural killer (NK) cell phenotype analyses from (A) fed, (B) 1-day-fasted and (C) 3-day-fasted mice are presented in dot plots and histograms. TCRβ− NK1.1+ cells were gated as NK cells. The dotted lines represent the negative control. The distribution of TRAIL, CD69, CD122, and CD25 expression in NK cells is indicated by the solid lines (shaded areas) and the percentage and mean fluorescence intensity (MFI) of positive cells are provided. (D) The percentages or (E) MFI of liver NK cells that are positive for TRAIL, CD69, CD122, and CD25 are shown in bar graphs as mean plus standard deviation. (F) Dot plots of representative data and (G) bar graph present the mean plus standard deviation of proportion of NK cell subsets regarding to TRAIL and CD69 expression in fed and fasted mice; *p <0.05, **p <0.01 as analyzed by the independent samples T test.
Mentions: Electronically gated TCRβ− NK1.1+ NK cells and NK cell markers from a representative fed, 1-day-fasted, or 3-day-fasted mouse are shown in Figure 1 A–C. Notably, compared to fed mice, 3-day-fasted mice showed significantly higher proportions of TRAIL and CD69 in liver NK cells (Figure 1D). Mean fluorescence intensity (MFI) of TRAIL or CD69 positive NK cells showed no significant differences among the groups (Figure 1E). Next, the distribution analysis of CD69 and TRAIL expression revealed that the proportion of CD69+TRAIL+ double positive NK cells significantly increased in fasted mice, while CD69− TRAIL− NK cells significantly decreased. The proportion of CD69+ TRAIL− cells also increased (Figure 1F, G). There was no difference in CD122 and CD25 expression in liver NK cells among the groups (Figure 1D, E) as well as in splenic NK cells (data not shown). The proportion of NK cells in the liver mononuclear cell fractions from 3-day-fasted mice did not differ from that from fed mice (Figure S1A).

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