Immune Cell Subtypes and Cytokines in Lung Tumor Microenvironment: Influence of COPD

Abstract

Background: The immune microenvironment plays a role in tumorigenesis. Chronic Obstructive Pulmonary Disease (COPD) is an independent risk factor for lung cancer (LC). We hypothesized that immune profile characterized by T regulatory (Treg), natural killer (NK), and plasma cells, as well as interleukin (IL)-10 and interferon-gamma, may differ within tumors of LC patients with/without COPD. Methods: Treg (anti-CD3 and anti-forkhead boxP3 antibodies), NK (anti-NCR1 antibody), IgG (anti-CD138-IgG antibody), IgA (anti-CD138-IgA antibody) using immunohistochemistry, and both IL-10 and interferon-gamma (ELISA) were quantified in tumor and non-tumor specimens (thoracotomy for lung tumor resection) from 33 LC–COPD patients and 20 LC-only patients. Results: Immune profile in tumor versus non-tumor specimens: Treg cell counts significantly increased in tumors of both LC and LC–COPD patients, while in tumors of the latter group, IgG-secreting plasma cells significantly decreased and IL-10 increased. No significant differences were seen in levels of NK cells, IgA-secreting cells, IgA/IgG, or interferon-gamma. Immune profile in tumors of LC–COPD versus LC: No significant differences were observed in tumors between LC–COPD and LC patients for any study marker. Conclusions: Immune cell subtypes and cytokines are differentially expressed in lung tumors, and the presence of COPD elicited a decline in IgG-secreting plasma cell levels but not in other cell types.

Increased PARP Activity and DNA Damage in NSCLC Patients: The Influence of COPD

“…DNA damage was assessed in tumor and non-tumor lung specimens by the presence of γ-H2AX, a hallmark of DNA damage [ 37 ] using conventional immunohistochemistry as previously described [ 10 , 38 ]. Briefly, paraffin-embedded specimens were cut into three-micrometer sections on a microtome.…”

“…PARP-1 and PARP-2 expression levels in tumor samples and non-tumor samples of human lungs were quantified using ELISA (MyBioSource, Inc., San Diego, CA, USA). All procedures were performed following the manufacturer’s instructions and previous studies [ 10 ]. The amount of total protein levels in lung homogenates were quantified using NanoDrop ND1000 spectrophotometer (Thermo Scientific, Waltham, MA, USA) with duplicates of each sample.…”

“…Another fragment was frozen immediately in liquid nitrogen and preserved at −80 • C for the measurement of protein levels. [37] using conventional immunohistochemistry as previously described [10,38]. Briefly, paraffin-embedded specimens were cut into three-micrometer sections on a microtome.…”

“…Underlying respiratory conditions favor the incidence of lung tumors in patients, especially in those with emphysema [ 8 , 9 ]. Mechanisms such as inflammation, oxidative stress, epigenetics alterations, and the tumor microenvironment underlie the pathophysiology of tumor development in patients with chronic respiratory disorders including stromal structures and immune cell profiles, and have an impact on the patients’ survival [ 10 , 11 , 12 ]. Indeed, chronic obstructive pulmonary disease (COPD) is a major risk factor for LC development [ 9 , 13 ].…”

B Cells and Tertiary Lymphoid Structures Influence Survival in Lung Cancer Patients with Resectable Tumors

“…B cells, T cells, and TLSs were identified on three-micrometer lung tumor and non-tumor cross-sections using double-staining immunohistochemical procedures (EnVision DuoFLEX Doublestain System, Dako North America Inc., Carpinteria, CA, USA) following the manufacturer’s instructions and previous study [ 10 , 44 , 45 ]. B and T cells were identified by staining of the lung samples with specific antibodies for B cells (anti-CD20 antibody, clone L26, Dako) and T cells (anti-CD3 antibody, Dako).…”