Caspase-3-independent apoptotic pathways contribute to interleukin-32γ-mediated control of Mycobacterium tuberculosis infection in THP-1 cells

BMC Microbiology

Edward D Chan⁵  Charles A Dinarello¹  Diane J Ordway²  Marcela Henao-Tamayo²  Mihai G Netea⁴  Jennifer R Honda⁵  Alida R Ovrutsky⁵  An Bai⁶  Wen-Lin Su³  William H Kinney⁵  Xiyuan Bai⁵ 

[1] Division of Infectious Diseases, University of Colorado Denver Anschutz Medical Campus, Aurora, CO, USA;Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO, USA;Division of Pulmonary and Critical Care Medicine, Taipei Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, New Taipei andTri-Service General Hospital; National Defense Medical Center, Taipei, Taiwan;Department of Internal Medicine and Radboud Center for Infectious Diseases, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands;Department of Medicine, University of Colorado School of Medicine, Aurora, CO, USA;Departments of Medicine and Academic Affairs, National Jewish Health, D509, Neustadt Building, 1400 Jackson Street, Denver 80206, CO, USA

关键词: Apoptosis-inducing factor;    Cathepsins;    Caspase-1;    Programmed cell death;    Apoptosis;    Interleukin-32;    Mycobacterium tuberculosis;   

Others  :  1137423
DOI  :  10.1186/s12866-015-0366-z

 received in 2014-10-29, accepted in 2015-01-29,  发布年份 2015

【 摘 要 】

Background

Macrophages are the primary effector cells responsible for killing Mycobacterium tuberculosis (MTB) through various mechanisms, including apoptosis. However, MTB can evade host immunity to create a favorable environment for intracellular replication. MTB-infected human macrophages produce interleukin-32 (IL-32). IL-32 is a pro-inflammatory cytokine and has several isoforms.We previously found that IL-32γ reduced the burden of MTB in human macrophages, in part, through the induction of caspase-3-dependent apoptosis. However, based on our previous studies, we hypothesized that caspase-3-independent death pathways may also mediate IL-32 control of MTB infection. Herein, we assessed the potential roles of cathepsin-mediated apoptosis, caspase-1-mediated pyroptosis, and apoptosis-inducing factor (AIF) in mediating IL-32γ control of MTB infection in THP-1 cells.

Results

Differentiated human THP-1 macrophages were infected with MTB H37Rv alone or in the presence of specific inhibitors to caspase-1, cathepsin B/D, or cathepsin L for up to four days, after which TUNEL-positive cells were quantified; in addition, MTB was quantified by culture as well as by the percentage of THP-1 cells that were infected with green fluorescent protein (GFP)-labeled MTB as determined by microscopy. AIF expression was inhibited using siRNA technology. Inhibition of cathepsin B/D, cathepsin L, or caspase-1 activity significantly abrogated the IL-32γ-mediated reduction in the number of intracellular MTB and of the percentage of GFP-MTB-infected macrophages. Furthermore, inhibition of caspase-1, cathepsin B/D, or cathepsin L in the absence of exogenous IL-32γ resulted in a trend toward an increased proportion of MTB-infected THP-1 cells.Inhibition of AIF activity in the absence of exogenous IL-32γ also increased intracellular burden of MTB. However, since IL-32γ did not induce AIF and because the relative increases in MTB with inhibition of AIF were similar in the presence or absence of IL-32γ, our results indicate that AIF does not mediate the host-protective effect of IL-32γ against MTB.

Conclusions

The anti-MTB effects of IL-32γ are mediated through classical caspase-3-dependent apoptosis as well as caspase-3-independent apoptosis.

【 授权许可】

   
2015 Bai et al.; licensee BioMed Central.

【 图 表 】

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