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Tumour microenvironment of both early- and late-stage colorectal cancer is equally immunosuppressive.

O'Toole A, Michielsen AJ, Nolan B, Tosetto M, Sheahan K, Mulcahy HE, Winter DC, Hyland JM, O'Connell PR, Fennelly D, O'Donoghue D, O'Sullivan J, Doherty GA, Ryan EJ - Br. J. Cancer (2014)

Bottom Line: The profile of inflammatory mediators in TCM was similar across stages, and all increased pSTAT3 expression by DCs.CRC patient DCs (n=31) secreted low levels of IL-12p70 and failed to upregulate expression of maturation markers in response to LPS.Furthermore, in vitro culture of autologous DCs with TCM did not change the hypo-responsiveness of patient DCs.Measures to reverse the negative influence of the TME on DCs will optimise cancer vaccines in both early- and late-stage CRC.

View Article: PubMed Central - PubMed

Affiliation: 1] Centre for Colorectal Disease, St Vincent's University Hospital, Dublin 4, Ireland [2] School of Medicine and Medical Sciences, University College Dublin, Dublin 4, Ireland.

ABSTRACT

Background: Tumour microenvironment (TME) of advanced colorectal cancer (CRC) suppresses dendritic cell (DC) maturation. Here, our aim was to determine how the microenvironment of early-stage tumours influences DCs.

Methods: Tumour-conditioned media (TCM) was generated by culturing explant tumour tissue in vitro (n=50). Monocyte-derived DCs (MDDCs) of healthy donors or cancer patients were pretreated with TCM and stimulated with lipopolysaccharide (LPS). DC maturation was assessed by flow cytometry and cytokine production measured by ELISA.

Results: TCM from both early- and late-staged tumours abrogated LPS-induction of IL-12p70 secretion, while increasing IL-10. The profile of inflammatory mediators in TCM was similar across stages, and all increased pSTAT3 expression by DCs.CRC patient DCs (n=31) secreted low levels of IL-12p70 and failed to upregulate expression of maturation markers in response to LPS. Furthermore, in vitro culture of autologous DCs with TCM did not change the hypo-responsiveness of patient DCs.

Conclusion: Our data demonstrates that the TME of all stages of CRC contains inflammatory mediators capable of suppressing local DCs. MDDCs obtained from CRC patients are hyporesponsive to stimuli such as LPS. Measures to reverse the negative influence of the TME on DCs will optimise cancer vaccines in both early- and late-stage CRC.

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Related in: MedlinePlus

TME of all colorectal cancer stages contains high levels of inflammatory mediators including IL-6. TCM of all stages of colorectal cancer was screened for the presence of a number of inflammatory mediators using an antibody array. High levels of IL-6, IL-8, GRO, angiogenin and TIMP1 were expressed in TCM of all stages (A). Levels of IL-6 were quantified by ELISA in TCM from all stages of CRC (n=5 per stage) (B). DCs isolated from healthy controls were stimulated for 15 min with IL-6 (100 ng ml−1) or cultured with a 1 : 2 dilution of TCM of stage I or stage IV TCM. The levels of pSTAT3 were analysed by flow cytometry (C).
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fig2: TME of all colorectal cancer stages contains high levels of inflammatory mediators including IL-6. TCM of all stages of colorectal cancer was screened for the presence of a number of inflammatory mediators using an antibody array. High levels of IL-6, IL-8, GRO, angiogenin and TIMP1 were expressed in TCM of all stages (A). Levels of IL-6 were quantified by ELISA in TCM from all stages of CRC (n=5 per stage) (B). DCs isolated from healthy controls were stimulated for 15 min with IL-6 (100 ng ml−1) or cultured with a 1 : 2 dilution of TCM of stage I or stage IV TCM. The levels of pSTAT3 were analysed by flow cytometry (C).

Mentions: TCM generated from CRC stages I–IV were screened for the presence of a panel of inflammatory mediators using an antibody based protein-array. Figure 2A shows that TCM of all stages of CRC progression contains a similar profile of inflammatory mediators. Of note, high levels of IL-6, IL-8, GRO and angiogenin are present. Using ELISA we confirmed that IL-6 was present in equivalent concentration in TCM from all tumour stages (Figure 2B).


Tumour microenvironment of both early- and late-stage colorectal cancer is equally immunosuppressive.

O'Toole A, Michielsen AJ, Nolan B, Tosetto M, Sheahan K, Mulcahy HE, Winter DC, Hyland JM, O'Connell PR, Fennelly D, O'Donoghue D, O'Sullivan J, Doherty GA, Ryan EJ - Br. J. Cancer (2014)

TME of all colorectal cancer stages contains high levels of inflammatory mediators including IL-6. TCM of all stages of colorectal cancer was screened for the presence of a number of inflammatory mediators using an antibody array. High levels of IL-6, IL-8, GRO, angiogenin and TIMP1 were expressed in TCM of all stages (A). Levels of IL-6 were quantified by ELISA in TCM from all stages of CRC (n=5 per stage) (B). DCs isolated from healthy controls were stimulated for 15 min with IL-6 (100 ng ml−1) or cultured with a 1 : 2 dilution of TCM of stage I or stage IV TCM. The levels of pSTAT3 were analysed by flow cytometry (C).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150274&req=5

fig2: TME of all colorectal cancer stages contains high levels of inflammatory mediators including IL-6. TCM of all stages of colorectal cancer was screened for the presence of a number of inflammatory mediators using an antibody array. High levels of IL-6, IL-8, GRO, angiogenin and TIMP1 were expressed in TCM of all stages (A). Levels of IL-6 were quantified by ELISA in TCM from all stages of CRC (n=5 per stage) (B). DCs isolated from healthy controls were stimulated for 15 min with IL-6 (100 ng ml−1) or cultured with a 1 : 2 dilution of TCM of stage I or stage IV TCM. The levels of pSTAT3 were analysed by flow cytometry (C).
Mentions: TCM generated from CRC stages I–IV were screened for the presence of a panel of inflammatory mediators using an antibody based protein-array. Figure 2A shows that TCM of all stages of CRC progression contains a similar profile of inflammatory mediators. Of note, high levels of IL-6, IL-8, GRO and angiogenin are present. Using ELISA we confirmed that IL-6 was present in equivalent concentration in TCM from all tumour stages (Figure 2B).

Bottom Line: The profile of inflammatory mediators in TCM was similar across stages, and all increased pSTAT3 expression by DCs.CRC patient DCs (n=31) secreted low levels of IL-12p70 and failed to upregulate expression of maturation markers in response to LPS.Furthermore, in vitro culture of autologous DCs with TCM did not change the hypo-responsiveness of patient DCs.Measures to reverse the negative influence of the TME on DCs will optimise cancer vaccines in both early- and late-stage CRC.

View Article: PubMed Central - PubMed

Affiliation: 1] Centre for Colorectal Disease, St Vincent's University Hospital, Dublin 4, Ireland [2] School of Medicine and Medical Sciences, University College Dublin, Dublin 4, Ireland.

ABSTRACT

Background: Tumour microenvironment (TME) of advanced colorectal cancer (CRC) suppresses dendritic cell (DC) maturation. Here, our aim was to determine how the microenvironment of early-stage tumours influences DCs.

Methods: Tumour-conditioned media (TCM) was generated by culturing explant tumour tissue in vitro (n=50). Monocyte-derived DCs (MDDCs) of healthy donors or cancer patients were pretreated with TCM and stimulated with lipopolysaccharide (LPS). DC maturation was assessed by flow cytometry and cytokine production measured by ELISA.

Results: TCM from both early- and late-staged tumours abrogated LPS-induction of IL-12p70 secretion, while increasing IL-10. The profile of inflammatory mediators in TCM was similar across stages, and all increased pSTAT3 expression by DCs.CRC patient DCs (n=31) secreted low levels of IL-12p70 and failed to upregulate expression of maturation markers in response to LPS. Furthermore, in vitro culture of autologous DCs with TCM did not change the hypo-responsiveness of patient DCs.

Conclusion: Our data demonstrates that the TME of all stages of CRC contains inflammatory mediators capable of suppressing local DCs. MDDCs obtained from CRC patients are hyporesponsive to stimuli such as LPS. Measures to reverse the negative influence of the TME on DCs will optimise cancer vaccines in both early- and late-stage CRC.

Show MeSH
Related in: MedlinePlus