Effects of cigarette smoke extract on primary activated T cells

doi:10.1016/j.cellimm.2013.04.005

Cellular Immunology

Volume 282, Issue 1, March 2013, Pages 38-43

https://doi.org/10.1016/j.cellimm.2013.04.005 Get rights and content

Highlights

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Cigarette smoke extract (CSE) blocks T cell proliferation by inducing apoptosis.
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Reactive nitrogen and oxygen species from CSE are responsible for T cell apoptosis.
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Phosphorylation of eIF2α is the central mediator for T cell apoptosis by CSE.

Abstract

Tobacco smoking predisposes the development of diseases characterized by chronic inflammation and T cell dysfunction. In this study, we aimed to determine the direct effects of cigarette smoke on primary T cells and to identify the corresponding molecular mediators. Activated T cells cultured in the presence of cigarette smoke extract (CSE) displayed a dose-dependent decrease in cell proliferation, which associated with the induction of cellular apoptosis. T cell apoptosis by CSE was independent of caspases and mediated through reactive oxygen and nitrogen species endogenously contained within CSE. Additional results showed that exposure of T cells to CSE induced phosphorylation of the stress mediator eukaryotic-translation-initiation-factor 2 alpha (eIF2α). Inhibition of the phosphorylation of eIF2α in T cells prevented the cellular apoptosis induced by CSE. Altogether, the results show the direct effects of CSE on T cells, which advance in the understanding of how cigarette smoking promotes chronic inflammation and immune dysfunction.

Introduction

Cigarette smoking is responsible for about 90% of the lung carcinoma cases worldwide [1] and represents a major risk factor for the development of several diseases including chronic obstructive pulmonary disease (COPD), asthma, coronary disease, and cancer [2], [3], [4], [5]. These conditions are characterized by the chronic presence of inflammatory immune responses [6], [7], suggesting a potential direct and/or indirect effect of cigarette smoking in immune cells [6]. Accordingly, previous studies have suggested that exposure to cigarette smoke leads to an increased susceptibility to infections such as influenza virus and mycobacterium tuberculosis and significant changes in cellular immune responses [8], [9]. However, the direct effects of cigarette smoke on T lymphocytes are still poorly understood.

The gaseous and particulate phases of cigarette smoke contain thousands of compounds, many of which are toxic agents. Among them, the generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) after tobacco burning play a major role in the cellular alterations induced by cigarette smoke on epithelial cells [10], [11]. In fact, previous studies had found an increased oxidative burden and a decreased in anti-oxidant pathways in the systemic compartment of smokers [12], [13].

Diverse stress signals including hypoxia, exposure to ultraviolet irradiation, and nutrient starvation, elicit in cells an integrated cellular response that is characterized by the phosphorylation of the eukaryotic translation initiation factor 2 alpha (eIF2α) [14]. Phosphorylation of eIF2α on serine residue 51 (phospho-eIF2α) inhibits nucleotide exchange on the eIF2 complex, attenuating cellular translation of most mRNAs and reducing protein synthesis [15], [14], [16]. During transient stress conditions, this process promotes the expression of proteins that function in the cellular adaptation to stress, which occurs through a Cap-independent translation [15], [14], [16]. However, if the stress conditions are persistent or highly aggressive for the cell, the phosphorylation of eIF2α will lead to the expression of proteins promoting cellular apoptosis.

The use of cigarette smoke extract (CSE) in vitro is a standardized method to test the direct effects induced by cigarette smoke on cells [17], [18]. Therefore, in this study, we aimed to determine the effect of CSE on primary T cells in vitro. Our results suggest that activated T cells cultured in the presence of CSE displayed a dose-dependent decrease in cell proliferation, which associated with the induction of T cell apoptosis. The induction of apoptosis by CSE was independent of caspase activation and mediated through ROS and RNS endogenously contained within the CSE. Additional results showed that the exposure of T cells to CSE induced the phosphorylation of eIF2α. Inhibition of the phosphorylation of eIF2α in T cells prevented the cellular apoptosis induced by CSE. Altogether, the results suggest that exposure of activated T cells to CSE induces apoptosis in a phospho-eIF2α-dependent manner. These results advance in the understanding of how cigarette smoking modulates responses in immune cell populations.

Section snippets

Cells lines, animals, and reagents

Human primary T cells were isolated from peripheral blood mononuclear cells using T cell enrichment columns (R&D Systems, Minneapolis, MN), as previously reported [19]. T cell purity ranged between 90% and 95%. Samples from healthy donors were obtained from buffy coats purchased from blood banks (around 30 different blood units were used during the study). Then, T cells were activated as previously described [20]. Murine T cells were isolated from spleens and lymph nodes of C57BL/6 mice and gp91

T lymphocytes cultured in CSE have a decreased proliferation and high rate of cellular apoptosis

To determine the effect of cigarette smoke on T cells, we have used CSE, a standardized method to test the effects of cigarette smoke on cultured cells [17], [18]. Human activated T lymphocytes were cultured in the presence of increasing concentrations of CSE (1.25–5%) and cell proliferation tested after 72 h. A dose dependent inhibition in cell proliferation was observed in activated T lymphocytes cultured with CSE (Fig. 1A) (P < 0.0001). The anti-proliferative effect induced by CSE in T cells

Discussion

Cigarette smoking is the leading cause of death in the United States and a major risk for the development of cancer, heart and respiratory diseases. Because of the negative impact of cigarette smoke in health and the persistent damage after cessation, it is important to understand the mechanisms by which cigarette smoke exposure induces cell damage. Our results suggest that ROS/RNS contained within CSE lead to apoptosis in primary T cells through caspase-independent mechanisms that are mediated

Acknowledgment

We thank Dr. Jey Jeyaseelan, PhD (LSU, Baton Rouge) for their input, help, and support during this work.

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^☆: This work was supported in part by NIH-NCRR (COBRE) P20RR021970 to P.C.R. and 1R21CA162133 to PCR.

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Effects of cigarette smoke extract on primary activated T cells☆