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Local and systemic neutrophilic inflammation in patients with lung cancer and chronic obstructive pulmonary disease.

Vaguliene N, Zemaitis M, Lavinskiene S, Miliauskas S, Sakalauskas R - BMC Immunol. (2013)

Bottom Line: NE and MPO levels in the serum and BAL fluid were determined by ELISA.ROS production was analyzed by flow cytometer.However, BAL fluid and serum levels of both NE and MPO were significantly higher in patients with lung cancer than COPD patients or healthy individuals (P < 0.05).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pulmonology and Immunology, Medical Academy, Hospital of Lithuanian University of Health Sciences, Eiveniu 2, Kaunas LT-50028, Lithuania. neringa.vaguliene@gmail.com

ABSTRACT

Background: Recent investigations suggest that neutrophils play an important role in the immune response to lung cancer as well as chronic obstructive pulmonary disease (COPD). The aim of this study was to evaluate the amount of neutrophils and markers of their activity in lung cancer and COPD and in coexistence of these two diseases.

Methods: In total, 267 persons were included in the study: 139 patients with lung cancer, 55 patients with lung cancer and COPD, 40 patients with COPD, and 33 healthy subjects. Peripheral blood and BAL fluid samples were obtained for cell count analysis and determination of NE, MPO levels and ROS production. NE and MPO levels in the serum and BAL fluid were determined by ELISA. ROS production was analyzed by flow cytometer.

Results: The percentage, cell count of neutrophils and neutrophil to lymphocyte ratio in the peripheral blood were significantly higher in lung cancer patients with or without COPD compared to COPD patients or healthy individuals (P < 0.05). The percentage and cell count of neutrophils in BAL fluid were significantly lower in patients with lung cancer with or without COPD than in patients with COPD (P < 0.05). However, BAL fluid and serum levels of both NE and MPO were significantly higher in patients with lung cancer than COPD patients or healthy individuals (P < 0.05). Neutrophils produced higher amounts of ROS in patients with lung cancer with or without COPD compared with COPD patients or healthy individuals (P < 0.05).

Conclusions: The results from this study demonstrate higher degree of local and systemic neutrophilic inflammation in patients with lung cancer (with or without COPD) than in patients with COPD.

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Related in: MedlinePlus

A histogram representing changes in ROS production in the neutrophils of peripheral blood of study patients after stimulation with different concentrations of PMA. Data are presented as mean fluorescence intensity (MFI) ± SEM. *P < 0.05, when compared with COPD and healthy individuals groups; §P < 0.05, when compared with healthy individuals. The representative dot plot of neurophil population isolated from peripheral blood of healthy individuals (A), patients with COPD (B), patients with lung cancer/COPD (C) and patients with lung cancer (D) stimulated upon 30 nM of Phorbol 12-myristate 13-acetate (PMA). Side light scater (SSC) represents the granularity, complexity of the cells; FSC – forward light scatter (FSC) represents cell size; dihydrorhodamine-123 (DHR-123) a green fluorescent compound showing H2O2 intensity in neutrophils.
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Figure 1: A histogram representing changes in ROS production in the neutrophils of peripheral blood of study patients after stimulation with different concentrations of PMA. Data are presented as mean fluorescence intensity (MFI) ± SEM. *P < 0.05, when compared with COPD and healthy individuals groups; §P < 0.05, when compared with healthy individuals. The representative dot plot of neurophil population isolated from peripheral blood of healthy individuals (A), patients with COPD (B), patients with lung cancer/COPD (C) and patients with lung cancer (D) stimulated upon 30 nM of Phorbol 12-myristate 13-acetate (PMA). Side light scater (SSC) represents the granularity, complexity of the cells; FSC – forward light scatter (FSC) represents cell size; dihydrorhodamine-123 (DHR-123) a green fluorescent compound showing H2O2 intensity in neutrophils.

Mentions: Neutrophils produced higher spontaneous ROS levels in the groups of lung cancer patients with or without COPD compared to the COPD patients or healthy individuals (P < 0.05) (Figure 1). The spontaneous ROS production in the lung cancer patients did not significantly differ despite the coexistence of COPD. Spontaneous ROS production in neutrophils did not differ between the male and female patients with lung cancer (175.53 ± 2.12 MFI vs. 175.29 ± 3.78 MFI, P > 0.05). There were no significant differences of spontaneous ROS production in neutrophils among the major histologic types of lung cancer: squamous cell carcinoma, adenocarcinoma, large cell carcinoma, non-small cell lung cancer not otherwise specified and small cell carcinoma (166.53 ± 4.93 MFI, 176.50 ± 2.73 MFI, 172.29 ± 4.56 MFI, 182.47 ± 6.43 MFI, 180.53 ± 3.98 MFI, P > 0.05) as well as between non-small cell lung cancer and small cell lung cancer groups (174.67 ± 2.02 MFI vs. 180.53 ± 3.98 MFI, P > 0.05). Additionally, spontaneous ROS production in neutrophils was significantly higher in patients with advanced lung cancer than in those with early lung cancer (183.66 ± 1.78 MFI vs. 145.91 ± 2.67 MFI, P < 0.001) and in the lung cancer patients with poor performance status 2–3 compared with those with performance status 0–1 (209.10 ± 4.93 MFI vs. 169.37 ± 1.52 MFI, P < 0.001). Furthermore, the patients with early lung cancer had a significantly higher spontaneous ROS production in neutrophils than the patients with COPD (P < 0.01). There were no significant differences of spontaneous ROS production in neutrophils in lung cancer groups among never smokers, former and current smokers (174.75 ± 4.93 MFI, 181.00 ± 5.65 MFI, 174.33 ± 2.19 MFI, P > 0.05). Additionally, spontaneous ROS production was found to be higher in lung cancer patients with or without COPD, who have never smoked, when comparing to current smokers with COPD (172.86 ± 5.26 and 179.25 ± 3.68 vs. 64.29 ± 1.17, P < 0.001). Different concentrations of PMA (0.3-30 nM) stimulated ROS production in neutrophils in all studied groups (Figure 1). An obvious increase of ROS production in neutrophils was detected after stimulation with 0,3 nM of PMA in lung cancer patients (with and without COPD), and with 10 nM in COPD group (P < 0.05). But the most significant increase of ROS production in all studied groups was observed in neutrophils stimulated with 30 nM PMA. There were no correlations between spontaneous ROS production in neutrophils and age, BMI and smoking intensity in study groups (data not shown).


Local and systemic neutrophilic inflammation in patients with lung cancer and chronic obstructive pulmonary disease.

Vaguliene N, Zemaitis M, Lavinskiene S, Miliauskas S, Sakalauskas R - BMC Immunol. (2013)

A histogram representing changes in ROS production in the neutrophils of peripheral blood of study patients after stimulation with different concentrations of PMA. Data are presented as mean fluorescence intensity (MFI) ± SEM. *P < 0.05, when compared with COPD and healthy individuals groups; §P < 0.05, when compared with healthy individuals. The representative dot plot of neurophil population isolated from peripheral blood of healthy individuals (A), patients with COPD (B), patients with lung cancer/COPD (C) and patients with lung cancer (D) stimulated upon 30 nM of Phorbol 12-myristate 13-acetate (PMA). Side light scater (SSC) represents the granularity, complexity of the cells; FSC – forward light scatter (FSC) represents cell size; dihydrorhodamine-123 (DHR-123) a green fluorescent compound showing H2O2 intensity in neutrophils.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750549&req=5

Figure 1: A histogram representing changes in ROS production in the neutrophils of peripheral blood of study patients after stimulation with different concentrations of PMA. Data are presented as mean fluorescence intensity (MFI) ± SEM. *P < 0.05, when compared with COPD and healthy individuals groups; §P < 0.05, when compared with healthy individuals. The representative dot plot of neurophil population isolated from peripheral blood of healthy individuals (A), patients with COPD (B), patients with lung cancer/COPD (C) and patients with lung cancer (D) stimulated upon 30 nM of Phorbol 12-myristate 13-acetate (PMA). Side light scater (SSC) represents the granularity, complexity of the cells; FSC – forward light scatter (FSC) represents cell size; dihydrorhodamine-123 (DHR-123) a green fluorescent compound showing H2O2 intensity in neutrophils.
Mentions: Neutrophils produced higher spontaneous ROS levels in the groups of lung cancer patients with or without COPD compared to the COPD patients or healthy individuals (P < 0.05) (Figure 1). The spontaneous ROS production in the lung cancer patients did not significantly differ despite the coexistence of COPD. Spontaneous ROS production in neutrophils did not differ between the male and female patients with lung cancer (175.53 ± 2.12 MFI vs. 175.29 ± 3.78 MFI, P > 0.05). There were no significant differences of spontaneous ROS production in neutrophils among the major histologic types of lung cancer: squamous cell carcinoma, adenocarcinoma, large cell carcinoma, non-small cell lung cancer not otherwise specified and small cell carcinoma (166.53 ± 4.93 MFI, 176.50 ± 2.73 MFI, 172.29 ± 4.56 MFI, 182.47 ± 6.43 MFI, 180.53 ± 3.98 MFI, P > 0.05) as well as between non-small cell lung cancer and small cell lung cancer groups (174.67 ± 2.02 MFI vs. 180.53 ± 3.98 MFI, P > 0.05). Additionally, spontaneous ROS production in neutrophils was significantly higher in patients with advanced lung cancer than in those with early lung cancer (183.66 ± 1.78 MFI vs. 145.91 ± 2.67 MFI, P < 0.001) and in the lung cancer patients with poor performance status 2–3 compared with those with performance status 0–1 (209.10 ± 4.93 MFI vs. 169.37 ± 1.52 MFI, P < 0.001). Furthermore, the patients with early lung cancer had a significantly higher spontaneous ROS production in neutrophils than the patients with COPD (P < 0.01). There were no significant differences of spontaneous ROS production in neutrophils in lung cancer groups among never smokers, former and current smokers (174.75 ± 4.93 MFI, 181.00 ± 5.65 MFI, 174.33 ± 2.19 MFI, P > 0.05). Additionally, spontaneous ROS production was found to be higher in lung cancer patients with or without COPD, who have never smoked, when comparing to current smokers with COPD (172.86 ± 5.26 and 179.25 ± 3.68 vs. 64.29 ± 1.17, P < 0.001). Different concentrations of PMA (0.3-30 nM) stimulated ROS production in neutrophils in all studied groups (Figure 1). An obvious increase of ROS production in neutrophils was detected after stimulation with 0,3 nM of PMA in lung cancer patients (with and without COPD), and with 10 nM in COPD group (P < 0.05). But the most significant increase of ROS production in all studied groups was observed in neutrophils stimulated with 30 nM PMA. There were no correlations between spontaneous ROS production in neutrophils and age, BMI and smoking intensity in study groups (data not shown).

Bottom Line: NE and MPO levels in the serum and BAL fluid were determined by ELISA.ROS production was analyzed by flow cytometer.However, BAL fluid and serum levels of both NE and MPO were significantly higher in patients with lung cancer than COPD patients or healthy individuals (P < 0.05).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pulmonology and Immunology, Medical Academy, Hospital of Lithuanian University of Health Sciences, Eiveniu 2, Kaunas LT-50028, Lithuania. neringa.vaguliene@gmail.com

ABSTRACT

Background: Recent investigations suggest that neutrophils play an important role in the immune response to lung cancer as well as chronic obstructive pulmonary disease (COPD). The aim of this study was to evaluate the amount of neutrophils and markers of their activity in lung cancer and COPD and in coexistence of these two diseases.

Methods: In total, 267 persons were included in the study: 139 patients with lung cancer, 55 patients with lung cancer and COPD, 40 patients with COPD, and 33 healthy subjects. Peripheral blood and BAL fluid samples were obtained for cell count analysis and determination of NE, MPO levels and ROS production. NE and MPO levels in the serum and BAL fluid were determined by ELISA. ROS production was analyzed by flow cytometer.

Results: The percentage, cell count of neutrophils and neutrophil to lymphocyte ratio in the peripheral blood were significantly higher in lung cancer patients with or without COPD compared to COPD patients or healthy individuals (P < 0.05). The percentage and cell count of neutrophils in BAL fluid were significantly lower in patients with lung cancer with or without COPD than in patients with COPD (P < 0.05). However, BAL fluid and serum levels of both NE and MPO were significantly higher in patients with lung cancer than COPD patients or healthy individuals (P < 0.05). Neutrophils produced higher amounts of ROS in patients with lung cancer with or without COPD compared with COPD patients or healthy individuals (P < 0.05).

Conclusions: The results from this study demonstrate higher degree of local and systemic neutrophilic inflammation in patients with lung cancer (with or without COPD) than in patients with COPD.

Show MeSH
Related in: MedlinePlus