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1 alpha, 25-dihydroxylvitamin D3 promotes Bacillus Calmette-Guérin immunotherapy of bladder cancer.

Hsu JW, Yin PN, Wood R, Messing J, Messing E, Lee YF - Oncotarget (2013)

Bottom Line: However, the mechanisms of BCG action have not been completely understood, thereby limiting the improvement of BCG therapy.Vitamin D deficiency has been associated with a high risk of TB infection, and the beneficial effect of UV exposure in TB patients was proven to be mediated via activation of vitamin D signals of innate immune cells.This THP-1 cell migration promoted by 1,25-VD can be blocked by IL-8 neutralized antibody.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, University of Rochester, Rochester, New York, USA.

ABSTRACT
Bacillus Calmette-Guérin (BCG), a vaccine against tuberculosis(TB), has been used and proven to be one of the most effective treatments for non-muscle invasive bladder cancer (BCa). However, the mechanisms of BCG action have not been completely understood, thereby limiting the improvement of BCG therapy. Vitamin D deficiency has been associated with a high risk of TB infection, and the beneficial effect of UV exposure in TB patients was proven to be mediated via activation of vitamin D signals of innate immune cells. Thus, vitamin D signals might be involved in mediating BCG immunotherapy. To test this hypothesis, we examined the impact of 1 alpha, 25-dihydroxyvitamin D3 (1,25-VD) on BCG-induced response in BCa cells and macrophage cells. Our data revealed that 1,25-VD promotes BCG-induced interleukin 8 (IL-8) secretion by BCa cells, consequently inducing the migration of macrophage, THP-1. This THP-1 cell migration promoted by 1,25-VD can be blocked by IL-8 neutralized antibody. Furthermore, 1,25-VD increased BCG-induced expression of macrophage markers in THP-1 cell, and enhanced the BCG-induced THP-1 cytotoxicity against low-grade BCa cells. Importantly, a pre-clinical trial using the N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN)-induced BCa mouse model revealed that intravesical co-treatment of 1,25-VD with BCG can prolong mice survival. These data demonstrate a novel mechanism by which 1,25-VD promotes BCG-mediated anti-BCa pathways and provides a platform for improving BCG efficacy with combination of 1,25-VD.

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1,25-VD promotes BCG induced THP-1 differentation/maturation(A) THP-1 cells were treated with vehicle, 1.25-VD, BCG, or combination overnight. Cells were collected for total RNA extraction. The gene expression levels of macrophage markers, CD163, CHIT1, and CH68 were determined by real-time PCR assay. (B) Representative photos display a morphological alteration after treatments were shown. (C) The expression of IL-8 after the treatment was detected by the real-time PCR and ELISA assays.
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Figure 4: 1,25-VD promotes BCG induced THP-1 differentation/maturation(A) THP-1 cells were treated with vehicle, 1.25-VD, BCG, or combination overnight. Cells were collected for total RNA extraction. The gene expression levels of macrophage markers, CD163, CHIT1, and CH68 were determined by real-time PCR assay. (B) Representative photos display a morphological alteration after treatments were shown. (C) The expression of IL-8 after the treatment was detected by the real-time PCR and ELISA assays.

Mentions: Vitamin D signaling is known to be critical for controlling mycobacteria infection through the regulation of TLR signal in innate immune cells, such as macrophage. Given the critical roles of macrophage in initiating an effective response to BCG therapy, we determined the expression of activated macrophage markers and cytokine secretion in THP-1 cells in responses to BCG and/or 1,25-VD. Three activated macrophage markers, CD163, Chitotriosidase (CHIT) and CD68, were examined when THP-1 cells were treated with BCG and/or 1,25-VD. We found that all three macrophage markers were significantly induced by BCG treatment. CD163 and CHIT, but not CD68, expression levels were further enhanced by additional 1,25-VD treatment (Fig. 4) in THP-1 cells. With respect to the elevated macrophage marker expression, BCG also induces morphological alteration in THP-1 cells where the majority of BGC-treated THP-1 cells were attached with a spindle-like morphology as compared to undifferentiated state where cells are round and suspended as shown in vehicle treated cells. Importantly, we found more spindle-like cells were attached suggesting 1,25-VD promotes BCG-induced macrophage differentiation; unexpectedly, we did not see too much alteration in the 1,25-VD treated THP-1 cells.


1 alpha, 25-dihydroxylvitamin D3 promotes Bacillus Calmette-Guérin immunotherapy of bladder cancer.

Hsu JW, Yin PN, Wood R, Messing J, Messing E, Lee YF - Oncotarget (2013)

1,25-VD promotes BCG induced THP-1 differentation/maturation(A) THP-1 cells were treated with vehicle, 1.25-VD, BCG, or combination overnight. Cells were collected for total RNA extraction. The gene expression levels of macrophage markers, CD163, CHIT1, and CH68 were determined by real-time PCR assay. (B) Representative photos display a morphological alteration after treatments were shown. (C) The expression of IL-8 after the treatment was detected by the real-time PCR and ELISA assays.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3926835&req=5

Figure 4: 1,25-VD promotes BCG induced THP-1 differentation/maturation(A) THP-1 cells were treated with vehicle, 1.25-VD, BCG, or combination overnight. Cells were collected for total RNA extraction. The gene expression levels of macrophage markers, CD163, CHIT1, and CH68 were determined by real-time PCR assay. (B) Representative photos display a morphological alteration after treatments were shown. (C) The expression of IL-8 after the treatment was detected by the real-time PCR and ELISA assays.
Mentions: Vitamin D signaling is known to be critical for controlling mycobacteria infection through the regulation of TLR signal in innate immune cells, such as macrophage. Given the critical roles of macrophage in initiating an effective response to BCG therapy, we determined the expression of activated macrophage markers and cytokine secretion in THP-1 cells in responses to BCG and/or 1,25-VD. Three activated macrophage markers, CD163, Chitotriosidase (CHIT) and CD68, were examined when THP-1 cells were treated with BCG and/or 1,25-VD. We found that all three macrophage markers were significantly induced by BCG treatment. CD163 and CHIT, but not CD68, expression levels were further enhanced by additional 1,25-VD treatment (Fig. 4) in THP-1 cells. With respect to the elevated macrophage marker expression, BCG also induces morphological alteration in THP-1 cells where the majority of BGC-treated THP-1 cells were attached with a spindle-like morphology as compared to undifferentiated state where cells are round and suspended as shown in vehicle treated cells. Importantly, we found more spindle-like cells were attached suggesting 1,25-VD promotes BCG-induced macrophage differentiation; unexpectedly, we did not see too much alteration in the 1,25-VD treated THP-1 cells.

Bottom Line: However, the mechanisms of BCG action have not been completely understood, thereby limiting the improvement of BCG therapy.Vitamin D deficiency has been associated with a high risk of TB infection, and the beneficial effect of UV exposure in TB patients was proven to be mediated via activation of vitamin D signals of innate immune cells.This THP-1 cell migration promoted by 1,25-VD can be blocked by IL-8 neutralized antibody.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, University of Rochester, Rochester, New York, USA.

ABSTRACT
Bacillus Calmette-Guérin (BCG), a vaccine against tuberculosis(TB), has been used and proven to be one of the most effective treatments for non-muscle invasive bladder cancer (BCa). However, the mechanisms of BCG action have not been completely understood, thereby limiting the improvement of BCG therapy. Vitamin D deficiency has been associated with a high risk of TB infection, and the beneficial effect of UV exposure in TB patients was proven to be mediated via activation of vitamin D signals of innate immune cells. Thus, vitamin D signals might be involved in mediating BCG immunotherapy. To test this hypothesis, we examined the impact of 1 alpha, 25-dihydroxyvitamin D3 (1,25-VD) on BCG-induced response in BCa cells and macrophage cells. Our data revealed that 1,25-VD promotes BCG-induced interleukin 8 (IL-8) secretion by BCa cells, consequently inducing the migration of macrophage, THP-1. This THP-1 cell migration promoted by 1,25-VD can be blocked by IL-8 neutralized antibody. Furthermore, 1,25-VD increased BCG-induced expression of macrophage markers in THP-1 cell, and enhanced the BCG-induced THP-1 cytotoxicity against low-grade BCa cells. Importantly, a pre-clinical trial using the N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN)-induced BCa mouse model revealed that intravesical co-treatment of 1,25-VD with BCG can prolong mice survival. These data demonstrate a novel mechanism by which 1,25-VD promotes BCG-mediated anti-BCa pathways and provides a platform for improving BCG efficacy with combination of 1,25-VD.

Show MeSH
Related in: MedlinePlus