Nasal lavage natural killer cell function is suppressed in smokers after live attenuated influenza virus

Horvath, Katherine M.; Herbst, M; Zhou, Haibo; Zhang, Hongtao; Noah, Terry L.; Jaspers, Ilona

doi:10.1186/1465-9921-12-102

Cited by 42 publications

(64 citation statements)

References 54 publications

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“…Thus although the anti-NE activity of SLPI is elevated in smokers, secreted SLPI levels in smokers may be altered and nonfunctional in the context of inflammation or infection. This is an important area of study considering that cigarette smoking results in proinflammatory events and impairs anti-IAV response both in vitro and in vivo (20,22,36).…”

Section: Discussionmentioning

confidence: 99%

“…Fourteen healthy young adults (9 men, 5 women); 7 nonsmokers (29.2 Ϯ 7.2 yr old) and 7 smokers (27.1 Ϯ 3.3 yr old), as characterized by our previous studies, were recruited to participate in this study (20). Informed consent was obtained from all subjects and the protocol was approved by the University of North Carolina Biomedical Institutional Review Board.…”

Section: Methodsmentioning

confidence: 99%

“…Formalin-fixed lungs were embedded in paraffin, cut into sections, and prepared as previously described (20). Slides were incubated with SLPI antibody (1:200; Santa Cruz) followed by incubation with a biotin-labeled anti-rabbit antibody, washed, incubated with avidin-biotin complex, and washed.…”

Section: Methodsmentioning

confidence: 99%

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Regulation and activity of secretory leukoprotease inhibitor (SLPI) is altered in smokers

Meyer

Bauer

Letang

et al. 2014

American Journal of Physiology-Lung Cellular and Molecular Phys

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A hallmark of cigarette smoking is a shift in the protease/antiprotease balance, in favor of protease activity. However, it has recently been shown that smokers have increased expression of a key antiprotease, secretory leukoprotease inhibitor (SLPI), yet the mechanisms involved in SLPI transcriptional regulation and functional activity of SLPI remain unclear. We examined SLPI mRNA and protein secretion in differentiated nasal epithelial cells (NECs) and nasal lavage fluid (NLF) from nonsmokers and smokers and demonstrated that SLPI expression is increased in NECs and NLF from smokers. Transcriptional regulation of SLPI expression was confirmed using SLPI promoter reporter assays followed by chromatin immunoprecipitation. The role of STAT1 in regulating SLPI expression was further elucidated using WT and stat1(-/-) mice. Our data demonstrate that STAT1 regulates SLPI transcription in epithelial cells and slpi protein in the lungs of mice. Additionally, we reveal that NECs from smokers have increased STAT1 mRNA/protein expression. Finally, we demonstrate that SLPI contained in the nasal mucosa of smokers is proteolytically cleaved but retains functional activity against neutrophil elastase. These results demonstrate that smoking enhances expression of SLPI in NECs in vitro and in vivo, and that this response is regulated by STAT1. In addition, despite posttranslational cleavage of SLPI, antiprotease activity against neutrophil elastase is enhanced in smokers. Together, our findings show that SLPI regulation and activity is altered in the nasal mucosa of smokers, which could have broad implications in the context of respiratory inflammation and infection.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Regulation and activity of secretory leukoprotease inhibitor (SLPI) is altered in smokers

Meyer

Bauer

Letang

et al. 2014

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…We and others have previously demonstrated that CS exposure significantly alters interferon-dependent antiviral responses at the level of the epithelium (19,40). Exposure to CS also alters the ability of epithelial cells to communicate with resident immune cells, such as dendritic cells and Natural Killer (NK) cells in the context of viral infections (17,18). Activation and maturation of NK cells can be modulated by soluble factors released by epithelial cells, such as cytokines/ chemokines, as well as ligands expressed on epithelial cells directly interacting with NK cell surface receptors (5).…”

mentioning

confidence: 99%

DNA methylation in nasal epithelial cells from smokers: identification of ULBP3-related effects

Rager¹,

Bauer²,

Müller

et al. 2013

American Journal of Physiology-Lung Cellular and Molecular Phys

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-We previously demonstrated that, in nasal epithelial cells (NECs) from smokers, methylation of an antiviral gene was associated with impaired antiviral defense responses. To expand these findings and better understand biological mechanisms underlying cigarette smoke (CS)-induced modifications of host defense responses, we aimed to compare DNA methylation of genes that may play a role in antiviral response. We used a two-tiered analytical approach, where we first implemented a genome-wide strategy. NECs from smokers differed in the methylation levels of 390 genes, the majority (84%) of which showed decreased methylation in smokers. Secondly, we generated an a priori set of 161 antiviral response-related genes, of which five were differentially methylated in NEC from smokers (CCL2, FDPS, GSK3B, SOCS3, and ULBP3). Assessing these genes at the systems biology level revealed a protein interaction network associated with CS-induced epigenetic modifications involving SOCS3 and ULBP3 signaling, among others. Subsequent confirmation studies focused on SOCS3 and ULBP3, which were hypomethylated and hypermethylated, respectively. Expression of SOCS3 was increased, whereas ULBP3 expression was decreased in NECs from smokers. Addition of the demethylating agent 5-Aza-2-deoxycytidine enhanced ULBP3 expression in NECs from smokers. Furthermore, infection of differentiated NECs with influenza virus resulted in significantly lower levels of ULBP3 in cells from smokers. Taken together, our findings show that genomic DNA methylation profiles are altered in NECs from smokers and that these changes are associated with decreased antiviral host defense responses, indicating that epigenenic dysregulation of genes such as SOCS3 and ULBP3 likely impacts immune responses in the epithelium.

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“…However, the less invasive procedure of scraping the inferior surface of the inferior nasal turbinate with a Rhino Probe can be performed in these subjects. Third, nasal scrape biopsies can be performed on the same individuals multiple times (usually about 4 weeks in between biopsies) without any significant side effects [28][29][30] . Fourth, NECs are among the first cells to be exposed to inhaled environmental stressors, such as air pollutants, pathogens or allergens and therefore represent valuable in vitro models to study the effects of these stressors on airway epithelial cells.…”

Section: Seed the Cells On Tissue Culture Insertsmentioning

confidence: 99%

Culturing of Human Nasal Epithelial Cells at the Air Liquid Interface

Müller

Brighton

Carson

et al. 2013

JoVE

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112

103

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In vitro models using human primary epithelial cells are essential in understanding key functions of the respiratory epithelium in the context of microbial infections or inhaled agents. Direct comparisons of cells obtained from diseased populations allow us to characterize different phenotypes and dissect the underlying mechanisms mediating changes in epithelial cell function. Culturing epithelial cells from the human tracheobronchial region has been well documented, but is limited by the availability of human lung tissue or invasiveness associated with obtaining the bronchial brushes biopsies. Nasal epithelial cells are obtained through much less invasive superficial nasal scrape biopsies and subjects can be biopsied multiple times with no significant side effects. Additionally, the nose is the entry point to the respiratory system and therefore one of the first sites to be exposed to any kind of air-borne stressor, such as microbial agents, pollutants, or allergens.Briefly, nasal epithelial cells obtained from human volunteers are expanded on coated tissue culture plates, and then transferred onto cell culture inserts. Upon reaching confluency, cells continue to be cultured at the air-liquid interface (ALI), for several weeks, which creates more physiologically relevant conditions. The ALI culture condition uses defined media leading to a differentiated epithelium that exhibits morphological and functional characteristics similar to the human nasal epithelium, with both ciliated and mucus producing cells. Tissue culture inserts with differentiated nasal epithelial cells can be manipulated in a variety of ways depending on the research questions (treatment with pharmacological agents, transduction with lentiviral vectors, exposure to gases, or infection with microbial agents) and analyzed for numerous different endpoints ranging from cellular and molecular pathways, functional changes, morphology, etc.In vitro models of differentiated human nasal epithelial cells will enable investigators to address novel and important research questions by using organotypic experimental models that largely mimic the nasal epithelium in vivo.

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Nasal lavage natural killer cell function is suppressed in smokers after live attenuated influenza virus

Cited by 42 publications

References 54 publications

Regulation and activity of secretory leukoprotease inhibitor (SLPI) is altered in smokers

Regulation and activity of secretory leukoprotease inhibitor (SLPI) is altered in smokers

DNA methylation in nasal epithelial cells from smokers: identification of ULBP3-related effects

Culturing of Human Nasal Epithelial Cells at the Air Liquid Interface

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