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The effect of lung cancer on cytokine expression in peripheral blood mononuclear cells.

Chang DH, Rutledge JR, Patel AA, Heerdt BG, Augenlicht LH, Korst RJ - PLoS ONE (2013)

Bottom Line: PBMC gene expression of CCL3, IL8 and IL1β was higher in lung cancer patients compared to the same patients at each of four sequential timepoints after removal of their tumors, while CXCL10 and IL2Rα were essentially unchanged.When non-cancer patients underwent surgery for benign diseases, these cytokine expression changes were not demonstrable.We conclude that PBMCs from stage I lung cancer patients possess distinct cytokine expression patterns compared to both non-cancer patients, and lung cancer patients following tumor removal.

View Article: PubMed Central - PubMed

Affiliation: Center for Cancer Research and Genomic Medicine, The Daniel and Gloria Blumenthal Cancer Center, Paramus, New Jersey, United States of America.

ABSTRACT
The purpose of this study is to evaluate cytokine expression by peripheral blood mononuclear cells (PBMC) from stage I lung cancer patients and to confirm these expression patterns by exposing PBMCs to lung cancer cells in vitro. Five altered cytokines in stage I lung cancer patients (CCL3, IL8, IL1β, CXCL10, sIL2Rα) were identified in plasma from subjects (n = 15) before and after resection using a 30-plex panel protein assay. Gene expression studies using quantitative RT-qPCR were performed on PBMCs from stage I lung cancer patients (n = 62) before and after resection, and compared to non-cancer patients (n = 32) before and after surgery for benign disease. Co-culture experiments that exposed healthy donor PBMCs to lung cancer cells in vitro were performed to evaluate the effect on PBMC cytokine expression. PBMC gene expression of CCL3, IL8 and IL1β was higher in lung cancer patients compared to the same patients at each of four sequential timepoints after removal of their tumors, while CXCL10 and IL2Rα were essentially unchanged. This pattern was also detected when lung cancer patients were compared to non-cancer patients. When non-cancer patients underwent surgery for benign diseases, these cytokine expression changes were not demonstrable. Lung cancer cell lines, but not benign bronchial epithelial cells, induced similar changes in cytokine gene and protein expression by healthy donor PBMCs in an in vitro co-culture system. We conclude that PBMCs from stage I lung cancer patients possess distinct cytokine expression patterns compared to both non-cancer patients, and lung cancer patients following tumor removal. These expression patterns are replicated by healthy donor PBMCs exposed to lung cancer cell lines, but not benign bronchial epithelial cells in vitro. These findings have implications for understanding the immune response to lung cancer.

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Summary of postoperative PBMC gene expression patterns for 5 selected cytokines.A repeated measures analysis of variance was used to isolate the effect of the presence or absence of lung cancer on the expression of CCL3 (p = 0.07), IL8 (p = 0.01), IL1β (p = 0.004), CXCL10 (p = 0.4) and IL2Rα (p = 0.69). Each bar represents the mean (+/− SEM) relative fold change for each cytokine incorporating all postoperative data compared to the expression before surgery, expressed in log2 scale. A negative relative fold change indicates downregulation after stage I lung cancer resection.
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pone-0064456-g002: Summary of postoperative PBMC gene expression patterns for 5 selected cytokines.A repeated measures analysis of variance was used to isolate the effect of the presence or absence of lung cancer on the expression of CCL3 (p = 0.07), IL8 (p = 0.01), IL1β (p = 0.004), CXCL10 (p = 0.4) and IL2Rα (p = 0.69). Each bar represents the mean (+/− SEM) relative fold change for each cytokine incorporating all postoperative data compared to the expression before surgery, expressed in log2 scale. A negative relative fold change indicates downregulation after stage I lung cancer resection.

Mentions: To validate and extend the results obtained from the initial Luminex and microarray data, PBMC gene expression was evaluated in much larger cohorts of stage I lung cancer patients (n = 62) and control patients undergoing thoracic surgery for benign diseases (n = 32). Demographic features of these two cohorts of patients are displayed in Table S2.The five selected target genes, CCL3, IL8, IL1β, CXCL10, and IL2Rα, were further investigated by the real-time RT-qPCR assay. Of the 62 patients in the lung cancer cohort who underwent phlebotomy prior to resection, 58 had blood drawn for analysis during the postoperative period, as did 16 of the 32 control patients. Figure 1 demonstrates the relative changes in expression detected for the 5 genes during the postoperative period for the lung cancer and control patients. Interestingly, PBMC gene expression for the three cytokines (CCL3, IL8, IL1β) detected in plasma at elevated levels prior to lung cancer resection during the initial screening were found to be downregulated after lung cancer resection at all timepoints. Further, the same pattern was not detected in the control patients undergoing thoracic surgery for benign diseases. Finally, PBMC gene expression for the 2 cytokines (CXCL10, IL2Rα) which were not detected in plasma at elevated levels prior to lung cancer resection during the initial screening were not significantly different after resection (Figure 1). Following analysis of these data using repeated measures ANOVA, it becomes clear that IL8 and IL1β expression are significantly downregulated after resection of stage I lung cancer, a finding which is not associated with the effect of thoracic surgery in non-cancer patients. CCL3 also appears to be downregulated in a similar fashion, to the extent that it approaches statistical significance (Figure 2). Further, flow cytometry of PBMCs shows a uniform population of lymphocyte subtypes across all patient cohorts, suggesting that changes in gene expression do not appear to be due to differences in cell representation in the PBMC lymphocyte populations, although these data do not eliminate the possibility of subtle shifts in subpopulations (Figure S2).


The effect of lung cancer on cytokine expression in peripheral blood mononuclear cells.

Chang DH, Rutledge JR, Patel AA, Heerdt BG, Augenlicht LH, Korst RJ - PLoS ONE (2013)

Summary of postoperative PBMC gene expression patterns for 5 selected cytokines.A repeated measures analysis of variance was used to isolate the effect of the presence or absence of lung cancer on the expression of CCL3 (p = 0.07), IL8 (p = 0.01), IL1β (p = 0.004), CXCL10 (p = 0.4) and IL2Rα (p = 0.69). Each bar represents the mean (+/− SEM) relative fold change for each cytokine incorporating all postoperative data compared to the expression before surgery, expressed in log2 scale. A negative relative fold change indicates downregulation after stage I lung cancer resection.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3675097&req=5

pone-0064456-g002: Summary of postoperative PBMC gene expression patterns for 5 selected cytokines.A repeated measures analysis of variance was used to isolate the effect of the presence or absence of lung cancer on the expression of CCL3 (p = 0.07), IL8 (p = 0.01), IL1β (p = 0.004), CXCL10 (p = 0.4) and IL2Rα (p = 0.69). Each bar represents the mean (+/− SEM) relative fold change for each cytokine incorporating all postoperative data compared to the expression before surgery, expressed in log2 scale. A negative relative fold change indicates downregulation after stage I lung cancer resection.
Mentions: To validate and extend the results obtained from the initial Luminex and microarray data, PBMC gene expression was evaluated in much larger cohorts of stage I lung cancer patients (n = 62) and control patients undergoing thoracic surgery for benign diseases (n = 32). Demographic features of these two cohorts of patients are displayed in Table S2.The five selected target genes, CCL3, IL8, IL1β, CXCL10, and IL2Rα, were further investigated by the real-time RT-qPCR assay. Of the 62 patients in the lung cancer cohort who underwent phlebotomy prior to resection, 58 had blood drawn for analysis during the postoperative period, as did 16 of the 32 control patients. Figure 1 demonstrates the relative changes in expression detected for the 5 genes during the postoperative period for the lung cancer and control patients. Interestingly, PBMC gene expression for the three cytokines (CCL3, IL8, IL1β) detected in plasma at elevated levels prior to lung cancer resection during the initial screening were found to be downregulated after lung cancer resection at all timepoints. Further, the same pattern was not detected in the control patients undergoing thoracic surgery for benign diseases. Finally, PBMC gene expression for the 2 cytokines (CXCL10, IL2Rα) which were not detected in plasma at elevated levels prior to lung cancer resection during the initial screening were not significantly different after resection (Figure 1). Following analysis of these data using repeated measures ANOVA, it becomes clear that IL8 and IL1β expression are significantly downregulated after resection of stage I lung cancer, a finding which is not associated with the effect of thoracic surgery in non-cancer patients. CCL3 also appears to be downregulated in a similar fashion, to the extent that it approaches statistical significance (Figure 2). Further, flow cytometry of PBMCs shows a uniform population of lymphocyte subtypes across all patient cohorts, suggesting that changes in gene expression do not appear to be due to differences in cell representation in the PBMC lymphocyte populations, although these data do not eliminate the possibility of subtle shifts in subpopulations (Figure S2).

Bottom Line: PBMC gene expression of CCL3, IL8 and IL1β was higher in lung cancer patients compared to the same patients at each of four sequential timepoints after removal of their tumors, while CXCL10 and IL2Rα were essentially unchanged.When non-cancer patients underwent surgery for benign diseases, these cytokine expression changes were not demonstrable.We conclude that PBMCs from stage I lung cancer patients possess distinct cytokine expression patterns compared to both non-cancer patients, and lung cancer patients following tumor removal.

View Article: PubMed Central - PubMed

Affiliation: Center for Cancer Research and Genomic Medicine, The Daniel and Gloria Blumenthal Cancer Center, Paramus, New Jersey, United States of America.

ABSTRACT
The purpose of this study is to evaluate cytokine expression by peripheral blood mononuclear cells (PBMC) from stage I lung cancer patients and to confirm these expression patterns by exposing PBMCs to lung cancer cells in vitro. Five altered cytokines in stage I lung cancer patients (CCL3, IL8, IL1β, CXCL10, sIL2Rα) were identified in plasma from subjects (n = 15) before and after resection using a 30-plex panel protein assay. Gene expression studies using quantitative RT-qPCR were performed on PBMCs from stage I lung cancer patients (n = 62) before and after resection, and compared to non-cancer patients (n = 32) before and after surgery for benign disease. Co-culture experiments that exposed healthy donor PBMCs to lung cancer cells in vitro were performed to evaluate the effect on PBMC cytokine expression. PBMC gene expression of CCL3, IL8 and IL1β was higher in lung cancer patients compared to the same patients at each of four sequential timepoints after removal of their tumors, while CXCL10 and IL2Rα were essentially unchanged. This pattern was also detected when lung cancer patients were compared to non-cancer patients. When non-cancer patients underwent surgery for benign diseases, these cytokine expression changes were not demonstrable. Lung cancer cell lines, but not benign bronchial epithelial cells, induced similar changes in cytokine gene and protein expression by healthy donor PBMCs in an in vitro co-culture system. We conclude that PBMCs from stage I lung cancer patients possess distinct cytokine expression patterns compared to both non-cancer patients, and lung cancer patients following tumor removal. These expression patterns are replicated by healthy donor PBMCs exposed to lung cancer cell lines, but not benign bronchial epithelial cells in vitro. These findings have implications for understanding the immune response to lung cancer.

Show MeSH
Related in: MedlinePlus