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Pancreatic ductal adenocarcinoma contains an effector and regulatory immune cell infiltrate that is altered by multimodal neoadjuvant treatment.

Shibuya KC, Goel VK, Xiong W, Sham JG, Pollack SM, Leahy AM, Whiting SH, Yeh MM, Yee C, Riddell SR, Pillarisetty VG - PLoS ONE (2014)

Bottom Line: Most intratumoral CD8+ T cells exhibited an antigen-experienced effector memory cell phenotype and were capable of producing IFN-γ.CD4+ regulatory T cells (Treg) and IL-17 producing T helper cells were significantly more prevalent in tumor than in blood.Consistent with the association with reduced survival in previous studies, we observed higher frequencies of both myeloid cells and Treg in poorly differentiated tumors.

View Article: PubMed Central - PubMed

Affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.

ABSTRACT

Objective: The immune response to pancreatic ductal adenocarcinoma (PDA) may play a role in defining its uniquely aggressive biology; therefore, we sought to clearly define the adaptive immune infiltrate in PDA.

Design: We used immunohistochemistry and flow cytometry to characterize the immune infiltrate in human PDA and compared our findings to the patients' peripheral blood.

Results: In contrast to the myeloid cell predominant infiltrate seen in murine models, T cells comprised the majority of the hematopoietic cell component of the tumor stroma in human PDA. Most intratumoral CD8+ T cells exhibited an antigen-experienced effector memory cell phenotype and were capable of producing IFN-γ. CD4+ regulatory T cells (Treg) and IL-17 producing T helper cells were significantly more prevalent in tumor than in blood. Consistent with the association with reduced survival in previous studies, we observed higher frequencies of both myeloid cells and Treg in poorly differentiated tumors. The majority of intratumoral T cells expressed the co-inhibitory receptor programmed death-1 (PD-1), suggesting one potential mechanism through which PDA may evade antitumor immunity. Successful multimodal neoadjuvant therapy altered the immunoregulatory balance and was associated with reduced infiltration of both myeloid cells and Treg.

Conclusion: Our data show that human PDA contains a complex mixture of inflammatory and regulatory immune cells, and that neoadjuvant therapy attenuates the infiltration of intratumoral cells associated with immunosuppression and worsened survival.

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T cells infiltrate PDA regardless of GM-CSF expression.A-C) IHC of normal adjacent pancreas, well-differentiated PDA, and poorly differentiated PDA showing dual staining of T cells (CD3+ in brown, arrows) and myeloid cells (CD11b+in purple, arrowheads). Open arrowhead indicates a normal duct. Photomicrographs taken with a 20x objective. D) Quantification of CD3+ and CD11b+ cells in PDA (p = 0.0009). E) Correlation between density of CD3+ and CD11b+ cells in each tumor (r = 0.51; p = 0.0008). F-H) Representative photomicrographs (40x) showing GM-CSF expression in normal pancreas and PDA. I) Distribution of myeloid cells and T cells stratified based upon chromogen intensity for GM-CSF. At least five 40x fields were analyzed per specimen. Scale bars = 25 µm.
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pone-0096565-g001: T cells infiltrate PDA regardless of GM-CSF expression.A-C) IHC of normal adjacent pancreas, well-differentiated PDA, and poorly differentiated PDA showing dual staining of T cells (CD3+ in brown, arrows) and myeloid cells (CD11b+in purple, arrowheads). Open arrowhead indicates a normal duct. Photomicrographs taken with a 20x objective. D) Quantification of CD3+ and CD11b+ cells in PDA (p = 0.0009). E) Correlation between density of CD3+ and CD11b+ cells in each tumor (r = 0.51; p = 0.0008). F-H) Representative photomicrographs (40x) showing GM-CSF expression in normal pancreas and PDA. I) Distribution of myeloid cells and T cells stratified based upon chromogen intensity for GM-CSF. At least five 40x fields were analyzed per specimen. Scale bars = 25 µm.

Mentions: We found that, while T cells and myeloid cells were rare in normal, non-neoplastic pancreatic parenchyma (Figure 1A), both cell types were present both within the stroma and adjacent to carcinoma cells throughout PDA tumors (Figure 1B&C). Since tumor differentiation in PDA has been shown to impact survival,[26] we categorized tumors based upon standard histological criteria into well- to moderately (well-mod) differentiated or poorly differentiated. We determined the average numbers of CD3+ and CD11b+ cells in regions containing carcinoma cells and found that T cells were significantly more prevalent than myeloid cells in both well-mod differentiated (p<0.0001) and poorly differentiated tumors (p = 0.03; Figure 1D). There were significantly more myeloid cells in poorly differentiated tumors (p = 0.04), as well as a trend towards an increase in T cell infiltrate (p = 0.06) based upon tumor differentiation status. Importantly, we discovered a strong positive correlation between the density of T cell and myeloid cell infiltration in our specimens (r = 0.51; p = 0.0008; Figure 1E).


Pancreatic ductal adenocarcinoma contains an effector and regulatory immune cell infiltrate that is altered by multimodal neoadjuvant treatment.

Shibuya KC, Goel VK, Xiong W, Sham JG, Pollack SM, Leahy AM, Whiting SH, Yeh MM, Yee C, Riddell SR, Pillarisetty VG - PLoS ONE (2014)

T cells infiltrate PDA regardless of GM-CSF expression.A-C) IHC of normal adjacent pancreas, well-differentiated PDA, and poorly differentiated PDA showing dual staining of T cells (CD3+ in brown, arrows) and myeloid cells (CD11b+in purple, arrowheads). Open arrowhead indicates a normal duct. Photomicrographs taken with a 20x objective. D) Quantification of CD3+ and CD11b+ cells in PDA (p = 0.0009). E) Correlation between density of CD3+ and CD11b+ cells in each tumor (r = 0.51; p = 0.0008). F-H) Representative photomicrographs (40x) showing GM-CSF expression in normal pancreas and PDA. I) Distribution of myeloid cells and T cells stratified based upon chromogen intensity for GM-CSF. At least five 40x fields were analyzed per specimen. Scale bars = 25 µm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4008589&req=5

pone-0096565-g001: T cells infiltrate PDA regardless of GM-CSF expression.A-C) IHC of normal adjacent pancreas, well-differentiated PDA, and poorly differentiated PDA showing dual staining of T cells (CD3+ in brown, arrows) and myeloid cells (CD11b+in purple, arrowheads). Open arrowhead indicates a normal duct. Photomicrographs taken with a 20x objective. D) Quantification of CD3+ and CD11b+ cells in PDA (p = 0.0009). E) Correlation between density of CD3+ and CD11b+ cells in each tumor (r = 0.51; p = 0.0008). F-H) Representative photomicrographs (40x) showing GM-CSF expression in normal pancreas and PDA. I) Distribution of myeloid cells and T cells stratified based upon chromogen intensity for GM-CSF. At least five 40x fields were analyzed per specimen. Scale bars = 25 µm.
Mentions: We found that, while T cells and myeloid cells were rare in normal, non-neoplastic pancreatic parenchyma (Figure 1A), both cell types were present both within the stroma and adjacent to carcinoma cells throughout PDA tumors (Figure 1B&C). Since tumor differentiation in PDA has been shown to impact survival,[26] we categorized tumors based upon standard histological criteria into well- to moderately (well-mod) differentiated or poorly differentiated. We determined the average numbers of CD3+ and CD11b+ cells in regions containing carcinoma cells and found that T cells were significantly more prevalent than myeloid cells in both well-mod differentiated (p<0.0001) and poorly differentiated tumors (p = 0.03; Figure 1D). There were significantly more myeloid cells in poorly differentiated tumors (p = 0.04), as well as a trend towards an increase in T cell infiltrate (p = 0.06) based upon tumor differentiation status. Importantly, we discovered a strong positive correlation between the density of T cell and myeloid cell infiltration in our specimens (r = 0.51; p = 0.0008; Figure 1E).

Bottom Line: Most intratumoral CD8+ T cells exhibited an antigen-experienced effector memory cell phenotype and were capable of producing IFN-γ.CD4+ regulatory T cells (Treg) and IL-17 producing T helper cells were significantly more prevalent in tumor than in blood.Consistent with the association with reduced survival in previous studies, we observed higher frequencies of both myeloid cells and Treg in poorly differentiated tumors.

View Article: PubMed Central - PubMed

Affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, United States of America.

ABSTRACT

Objective: The immune response to pancreatic ductal adenocarcinoma (PDA) may play a role in defining its uniquely aggressive biology; therefore, we sought to clearly define the adaptive immune infiltrate in PDA.

Design: We used immunohistochemistry and flow cytometry to characterize the immune infiltrate in human PDA and compared our findings to the patients' peripheral blood.

Results: In contrast to the myeloid cell predominant infiltrate seen in murine models, T cells comprised the majority of the hematopoietic cell component of the tumor stroma in human PDA. Most intratumoral CD8+ T cells exhibited an antigen-experienced effector memory cell phenotype and were capable of producing IFN-γ. CD4+ regulatory T cells (Treg) and IL-17 producing T helper cells were significantly more prevalent in tumor than in blood. Consistent with the association with reduced survival in previous studies, we observed higher frequencies of both myeloid cells and Treg in poorly differentiated tumors. The majority of intratumoral T cells expressed the co-inhibitory receptor programmed death-1 (PD-1), suggesting one potential mechanism through which PDA may evade antitumor immunity. Successful multimodal neoadjuvant therapy altered the immunoregulatory balance and was associated with reduced infiltration of both myeloid cells and Treg.

Conclusion: Our data show that human PDA contains a complex mixture of inflammatory and regulatory immune cells, and that neoadjuvant therapy attenuates the infiltration of intratumoral cells associated with immunosuppression and worsened survival.

Show MeSH
Related in: MedlinePlus