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Mesothelioma tumor cells modulate dendritic cell lipid content, phenotype and function.

Gardner JK, Mamotte CD, Patel P, Yeoh TL, Jackaman C, Nelson DJ - PLoS ONE (2015)

Bottom Line: Lipid accumulation was associated with reduced antigen processing ability (measured using a DQ OVA assay), upregulation of the co-stimulatory molecule, CD86, and production of the tolerogenic cytokine, IL-10.Moreover, increasing tumor burden was associated with reduced proliferation of tumor-antigen-specific CD8+ T cells in tumor-draining lymph nodes.This study shows that mesothelioma promotes DC lipid acquisition, which is associated with altered activation status and reduced capacity to process and present antigens, which may impair the ability of DCs to generate effective anti mesothelioma T cell responses.

View Article: PubMed Central - PubMed

Affiliation: Immunology and Cancer Group, School of Biomedical Sciences, Curtin University, Perth, Western Australia, Australia; CHIRI Biosciences Research Precinct, Curtin University, Perth, Western Australia, Australia.

ABSTRACT
Dendritic cells (DCs) play an important role in the generation of anti-cancer immune responses, however there is evidence that DCs in cancer patients are dysfunctional. Lipid accumulation driven by tumor-derived factors has recently been shown to contribute to DC dysfunction in several human cancers, but has not yet been examined in mesothelioma. This study investigated if mesothelioma tumor cells and/or their secreted factors promote increases in DC lipid content and modulate DC function. Human monocyte-derived DCs (MoDCs) were exposed to human mesothelioma tumor cells and tumor-derived factors in the presence or absence of lipoproteins. The data showed that immature MoDCs exposed to mesothelioma cells or factors contained increased lipid levels relative to control DCs. Lipid accumulation was associated with reduced antigen processing ability (measured using a DQ OVA assay), upregulation of the co-stimulatory molecule, CD86, and production of the tolerogenic cytokine, IL-10. Increases in DC lipid content were further enhanced by co-exposure to mesothelioma-derived factors and triglyceride-rich lipoproteins, but not low-density lipoproteins. In vivo studies using a murine mesothelioma model showed that the lipid content of tumor-infiltrating CD4+ CD8α- DCs, CD4- CD8α- DCs DCs and plasmacytoid DCs increased with tumor progression. Moreover, increasing tumor burden was associated with reduced proliferation of tumor-antigen-specific CD8+ T cells in tumor-draining lymph nodes. This study shows that mesothelioma promotes DC lipid acquisition, which is associated with altered activation status and reduced capacity to process and present antigens, which may impair the ability of DCs to generate effective anti mesothelioma T cell responses.

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Tumor-antigen-specific CD8+ T cell proliferation in draining lymph nodes decreases with increasing tumor burden.C57BL/6J mice were inoculated with 5 x105 AE17 tumor cells, which were used as negative controls (B) or with AE17sOVA tumor cells (growth rate shown in A; n = 10 mice; data shown as mean ± SEM) on day 0. CFSE-labelled, CD8+, class I restricted, OVA-specific T cells from OT-1 mice were adoptively transferred at days 4, 11, 18 and 25 into the tumor-bearing mice. The dLNs were harvested from recipient mice three days post transfer such that FACS analysis was on days 7 (C, G), 14 (D, G), 21 (E, G) and 28 (F, G) post tumor cell inoculation. The lymph nodes were prepared as single cell suspensions and stained for CD8 for re-isolation of CFSE-labelled OT-1 cells. FACS analysis was performed by gating on CD8+ T cells. Representative FACS profiles shown in B-F. Results from individual mice from 1 experiment with 3–6 mice/group (G).
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pone.0123563.g010: Tumor-antigen-specific CD8+ T cell proliferation in draining lymph nodes decreases with increasing tumor burden.C57BL/6J mice were inoculated with 5 x105 AE17 tumor cells, which were used as negative controls (B) or with AE17sOVA tumor cells (growth rate shown in A; n = 10 mice; data shown as mean ± SEM) on day 0. CFSE-labelled, CD8+, class I restricted, OVA-specific T cells from OT-1 mice were adoptively transferred at days 4, 11, 18 and 25 into the tumor-bearing mice. The dLNs were harvested from recipient mice three days post transfer such that FACS analysis was on days 7 (C, G), 14 (D, G), 21 (E, G) and 28 (F, G) post tumor cell inoculation. The lymph nodes were prepared as single cell suspensions and stained for CD8 for re-isolation of CFSE-labelled OT-1 cells. FACS analysis was performed by gating on CD8+ T cells. Representative FACS profiles shown in B-F. Results from individual mice from 1 experiment with 3–6 mice/group (G).

Mentions: The next experiments were designed to dissect out the in vivo relationship between tumor burden and tumor antigen presentation to CD8+ T cells in dLNs. A kinetic study was undertaken in which CFSE-labelled, CD8+ T cells prepared from the lymph nodes and spleens of OT-1 mice were adoptively transferred into recipient mice bearing either AE17 tumors (the negative control) or AE17sOVA tumors which express ovalbumin as a marker, or spy, tumor antigen and to which OT-1 T cells will respond by proliferating if the antigen is appropriately presented to them by local DCs [16]. The growth rate and tumor size of AE17sOVA is shown in Fig 10A. Throughout these experiments the proliferative responses of CFSE-labelled OT-1 cells in dLNs of recipient animals were analyzed 3 days post adoptive transfer. Responses were determined weekly over one month. A strong in vivo OT-1 proliferative response was seen in the dLNs of all mice bearing AE17sOVA tumors at days 7 and 14 (Fig 10C, 10D and 10G). This response started to diminish at day 21, disappearing by day 28 (Fig 10E, 10F and 10G). No OT-1 proliferation was seen in the dLNs at any time point in mice given control AE17 tumor cells (Fig 10B). These data show that decreasing tumor-antigen-specific CD8+ T cell proliferation in dLNs is associated with increasing tumor burden.


Mesothelioma tumor cells modulate dendritic cell lipid content, phenotype and function.

Gardner JK, Mamotte CD, Patel P, Yeoh TL, Jackaman C, Nelson DJ - PLoS ONE (2015)

Tumor-antigen-specific CD8+ T cell proliferation in draining lymph nodes decreases with increasing tumor burden.C57BL/6J mice were inoculated with 5 x105 AE17 tumor cells, which were used as negative controls (B) or with AE17sOVA tumor cells (growth rate shown in A; n = 10 mice; data shown as mean ± SEM) on day 0. CFSE-labelled, CD8+, class I restricted, OVA-specific T cells from OT-1 mice were adoptively transferred at days 4, 11, 18 and 25 into the tumor-bearing mice. The dLNs were harvested from recipient mice three days post transfer such that FACS analysis was on days 7 (C, G), 14 (D, G), 21 (E, G) and 28 (F, G) post tumor cell inoculation. The lymph nodes were prepared as single cell suspensions and stained for CD8 for re-isolation of CFSE-labelled OT-1 cells. FACS analysis was performed by gating on CD8+ T cells. Representative FACS profiles shown in B-F. Results from individual mice from 1 experiment with 3–6 mice/group (G).
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Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4401725&req=5

pone.0123563.g010: Tumor-antigen-specific CD8+ T cell proliferation in draining lymph nodes decreases with increasing tumor burden.C57BL/6J mice were inoculated with 5 x105 AE17 tumor cells, which were used as negative controls (B) or with AE17sOVA tumor cells (growth rate shown in A; n = 10 mice; data shown as mean ± SEM) on day 0. CFSE-labelled, CD8+, class I restricted, OVA-specific T cells from OT-1 mice were adoptively transferred at days 4, 11, 18 and 25 into the tumor-bearing mice. The dLNs were harvested from recipient mice three days post transfer such that FACS analysis was on days 7 (C, G), 14 (D, G), 21 (E, G) and 28 (F, G) post tumor cell inoculation. The lymph nodes were prepared as single cell suspensions and stained for CD8 for re-isolation of CFSE-labelled OT-1 cells. FACS analysis was performed by gating on CD8+ T cells. Representative FACS profiles shown in B-F. Results from individual mice from 1 experiment with 3–6 mice/group (G).
Mentions: The next experiments were designed to dissect out the in vivo relationship between tumor burden and tumor antigen presentation to CD8+ T cells in dLNs. A kinetic study was undertaken in which CFSE-labelled, CD8+ T cells prepared from the lymph nodes and spleens of OT-1 mice were adoptively transferred into recipient mice bearing either AE17 tumors (the negative control) or AE17sOVA tumors which express ovalbumin as a marker, or spy, tumor antigen and to which OT-1 T cells will respond by proliferating if the antigen is appropriately presented to them by local DCs [16]. The growth rate and tumor size of AE17sOVA is shown in Fig 10A. Throughout these experiments the proliferative responses of CFSE-labelled OT-1 cells in dLNs of recipient animals were analyzed 3 days post adoptive transfer. Responses were determined weekly over one month. A strong in vivo OT-1 proliferative response was seen in the dLNs of all mice bearing AE17sOVA tumors at days 7 and 14 (Fig 10C, 10D and 10G). This response started to diminish at day 21, disappearing by day 28 (Fig 10E, 10F and 10G). No OT-1 proliferation was seen in the dLNs at any time point in mice given control AE17 tumor cells (Fig 10B). These data show that decreasing tumor-antigen-specific CD8+ T cell proliferation in dLNs is associated with increasing tumor burden.

Bottom Line: Lipid accumulation was associated with reduced antigen processing ability (measured using a DQ OVA assay), upregulation of the co-stimulatory molecule, CD86, and production of the tolerogenic cytokine, IL-10.Moreover, increasing tumor burden was associated with reduced proliferation of tumor-antigen-specific CD8+ T cells in tumor-draining lymph nodes.This study shows that mesothelioma promotes DC lipid acquisition, which is associated with altered activation status and reduced capacity to process and present antigens, which may impair the ability of DCs to generate effective anti mesothelioma T cell responses.

View Article: PubMed Central - PubMed

Affiliation: Immunology and Cancer Group, School of Biomedical Sciences, Curtin University, Perth, Western Australia, Australia; CHIRI Biosciences Research Precinct, Curtin University, Perth, Western Australia, Australia.

ABSTRACT
Dendritic cells (DCs) play an important role in the generation of anti-cancer immune responses, however there is evidence that DCs in cancer patients are dysfunctional. Lipid accumulation driven by tumor-derived factors has recently been shown to contribute to DC dysfunction in several human cancers, but has not yet been examined in mesothelioma. This study investigated if mesothelioma tumor cells and/or their secreted factors promote increases in DC lipid content and modulate DC function. Human monocyte-derived DCs (MoDCs) were exposed to human mesothelioma tumor cells and tumor-derived factors in the presence or absence of lipoproteins. The data showed that immature MoDCs exposed to mesothelioma cells or factors contained increased lipid levels relative to control DCs. Lipid accumulation was associated with reduced antigen processing ability (measured using a DQ OVA assay), upregulation of the co-stimulatory molecule, CD86, and production of the tolerogenic cytokine, IL-10. Increases in DC lipid content were further enhanced by co-exposure to mesothelioma-derived factors and triglyceride-rich lipoproteins, but not low-density lipoproteins. In vivo studies using a murine mesothelioma model showed that the lipid content of tumor-infiltrating CD4+ CD8α- DCs, CD4- CD8α- DCs DCs and plasmacytoid DCs increased with tumor progression. Moreover, increasing tumor burden was associated with reduced proliferation of tumor-antigen-specific CD8+ T cells in tumor-draining lymph nodes. This study shows that mesothelioma promotes DC lipid acquisition, which is associated with altered activation status and reduced capacity to process and present antigens, which may impair the ability of DCs to generate effective anti mesothelioma T cell responses.

Show MeSH
Related in: MedlinePlus