Perforin Expression and Cytotoxic Activity of Sputum CD8+ Lymphocytes in Patients With COPD

doi:10.1378/chest.125.1.71

Chest

Volume 125, Issue 1, January 2004, Pages 71-76

https://doi.org/10.1378/chest.125.1.71 Get rights and content

Background

Previous studies have shown that the inflammatory response to cigarette smoking differs between smokers who acquire COPD and those who do not, and the CD8⁺ T- lymphocytes have been identified as a key player in this response.

Objective

To investigate the cytotoxic activity and perforin expression of CD8⁺ lymphocytes in the airway lumen of patients with COPD.

Methods

Thirty-six male smokers with COPD, 25 male smokers without COPD, and 10 healthy nonsmokers participated in the study. T-lymphocytes of induced sputum samples were labeled with appropriate monoclonal antibodies and measured using flow cytometry. The cytotoxic activity of CD8⁺ cells was defined by incubating them with specific target cells (K562).

Results

The percentage and the total number of CD8⁺ lymphocytes were significantly higher in COPD smokers compared to non-COPD smokers (p = 0.01 and p = 0.005, respectively) or to healthy nonsmokers (p = 0.02 and p = 0.01, respectively). Perforin expression in CD8⁺ cells was significantly higher in smokers with COPD compared to the other two groups (p = 0.001). Increased cytotoxic activity of T cells was also observed in induced sputum of patients with COPD in comparison to the other two groups.

Conclusion

CD8⁺ cells are not only increased in number in sputum samples of smokers with COPD but are highly activated, expressing high levels of perforin. These findings suggest that CD8⁺ T-lymphocytes play a significant role in the inflammatory process of COPD.

Section snippets

Subjects

Seventy-one male subjects were studied: 36 current smokers with COPD, 25 non-COPD smokers with a similar smoking history to the patients with COPD, and 10 healthy nonsmokers (Table 1). The diagnosis of COPD was based on the European consensus criteria.² All patients with COPD had been free of an acute exacerbation for at least 4 weeks preceding the study, and none had received antibiotics or corticosteroids (oral and inhaled) over the same period. Neither smokers nor healthy volunteers had a

Results

Demographic and spirometric data of the three groups of subjects are shown in Table 1. The patients with COPD exhibited significantly lower values of FEV₁ (percentage of predicted) and FEV₁/FVC (percentage) than non-COPD smokers, although the smoking history was similar. There were no significant differences in FEV₁/FVC (percentage) and FEV₁ (percentage) between non-COPD smokers and normal subjects. Change in FEV₁ (percentage of predicted) after salbutamol was similar in the three groups (Table

Discussion

In agreement with other studies,¹³¹⁸ we also showed that it is possible to analyze sputum samples for lymphocyte cell surface markers by flow cytometry. The main findings of the present study were that CD8⁺ lymphocytes of patients with COPD showed increased cytotoxic activity and increased expression of intracellular perforin compared to smokers without COPD, but with similar smoking history, and healthy nonsmokers (normal subjects). It is well known that it is difficult to induce sputum

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This study was supported by an unrestricted grant from ASTRA Hellas.

Part of this study has been presented at the 11th Hellenic Thoracic Society annual meeting, Thessalonica, December 6–9, 2001, and as prize winner an extensive summary has been published in the Greek journal Pneumon (2002; 15:78).

This work has been awarded the national prize during the 12th European Respiratory Society meeting, Stockholm, Sweden, September 14–18, 2002.

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Chest

Clinical InvestigationsCOPDPerforin Expression and Cytotoxic Activity of Sputum CD8+ Lymphocytes in Patients With COPD

Background

Objective

Methods

Results

Conclusion

Section snippets

Subjects

Results

Discussion

Respir Med

Blood

Chest

Immunol Today

Lancet

Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: National Heart, Lung, and Blood Institute and World Health Organization Global Initiative for Chronic Obstructive Lung Disease (GOLD); executive summary

Respir Care

Optimal assessment and management of chronic obstructive pulmonary disease (COPD). The European Respiratory Society Task Force

Eur Respir J

Chronic obstructive pulmonary disease

N Engl J Med

Inflammation in bronchial biopsies of subjects with chronic bronchitis: inverse relationship of CD8+ T lymphocytes with FEV1

Am J Respir Crit Care Med

CD8+ T-lymphocytes in peripheral airways of smokers with chronic obstructive pulmonary disease

Am J Respir Crit Care Med

Inflammatory reaction in pulmonary muscular arteries of patients with mild chronic obstructive pulmonary disease

Am J Respir Crit Care Med

The binding and lysis of target cells by cytotoxic lymphocytes: molecular and cellular aspects

Annu Rev Immunol

Perforin-independent CD8(+) T-cell-mediated cytotoxicity of alveolar epithelial cells is preferentially mediated by tumor necrosis factor-α: relative insensitivity to Fas ligand

Am J Respir Cell Mol Biol

Perforin-secreting killer cell infiltration and expression of a 65-kD heat-shock protein in aortic tissue of patients with Takayasu's arteritis

J Clin Invest

Activated CD4+ and CD8+ cytotoxic cells are present in increased numbers in the intestinal mucosa from patients with active inflammatory bowel disease

Am J Pathol

Clinical Investigations
COPD
Perforin Expression and Cytotoxic Activity of Sputum CD8⁺ Lymphocytes in Patients With COPD

Inflammation in bronchial biopsies of subjects with chronic bronchitis: inverse relationship of CD8+ T lymphocytes with FEV₁