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Brucella suis vaccine strain S2-infected immortalized caprine endometrial epithelial cell lines induce non-apoptotic ER-stress

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Cell Stress and Chaperones Aims and scope

Abstract

Brucella, which is regarded as an intracellular pathogen responsible for a zoonotic disease called brucellosis, survives and proliferates within several types of phagocytic and non-phagocytic cells. Brucella infects not only their preferred hosts but also other domestic and wild animal species, inducing abortion and infertility. Therefore, the interaction between uterine cells and Brucella is important for understanding the pathogenesis of this disease. In this study, we describe the Brucella suis vaccine strain S2 (B.suis.S2) infection and replication in the immortalized caprine endometrial epithelial cell line hTERT-EECs and the induced cellular and molecular response modulation in vitro. We found that B.suis S2 was able to infect and replicate to high titers and inhibit the proliferation of EECs and induce non-apoptotic pathways, as determined by B.suis.S2 detection using MTT and acridine orange/ethidium bromide (AO/EB) staining and flow cytometry. We explored the evidence of non-apoptotic pathways using real-time quantitative RT-PCR and by western blot analysis. Finally, we discovered the over-expression of GRP78, ATF4, ATF6, PERK, eIF2α, CHOP, and cytochrome c (Cyt-c) but not IRE1, xbp-1, and caspase-3 in B.suis.S2 (HK)-attacked and B.suis.S2-infected cells, suggesting that the molecular mechanism of ER stress sensor activation by B.suis.S2 is basically concomitant with that by B.suis.S2 (HK) and that ER stress, especially the PERK pathway, plays an important role in the process of B.suis.S2 infecting EEC, which may, in part, explain the role of the uterus in the pathogenesis of B.suis.S2.

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Abbreviations

T4SS:

Type IV secretion system

ER:

Endoplasmic reticulum

EEC:

Caprine endometrial epithelial cell

B.suis.S2 :

Brucella suis vaccine strain S2

PERK:

Protein kinase RNA-like kinase

IRE1α:

Inositol-requiring enzyme 1α

ATF6:

Activating transcription factor 6

GRP78:

78-kDa glucose-regulated protein

CHOP:

C/EBP homologus protein

CFU:

Colony-forming units

Tm:

Tunicamycin

xbp-1:

X-box binding protein 1

MTT-3-[4:

5-Dimethylthiazol-2-yl]-2

5:

Diphenyltetrazolium bromide assay

eIF2α:

α Subunit of eukaryotic initiation factor 2

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Acknowledgments

We thank the Shaanxi Provincial Institute for Veterinary Drug Control for vaccine strains.

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Authors and Affiliations

  1. Key Laboratory of Animal Biotechnology of the Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi, 712100, China

    Xiangguo Wang, Pengfei Lin, Yanlong Yin, Jinhua Zhou, Lanjie Lei, Xudong Zhou & Yaping Jin

  2. College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, 712100, China

    Xiangguo Wang, Pengfei Lin, Yanlong Yin, Jinhua Zhou, Lanjie Lei, Xudong Zhou, Yaping Jin & Aihua Wang

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  1. Xiangguo Wang
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  2. Pengfei Lin
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  3. Yanlong Yin
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  4. Jinhua Zhou
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  5. Lanjie Lei
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  6. Xudong Zhou
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  7. Yaping Jin
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  8. Aihua Wang
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Correspondence to Aihua Wang.

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Wang, X., Lin, P., Yin, Y. et al. Brucella suis vaccine strain S2-infected immortalized caprine endometrial epithelial cell lines induce non-apoptotic ER-stress. Cell Stress and Chaperones 20, 399–409 (2015). https://doi.org/10.1007/s12192-014-0564-x

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  • DOI: https://doi.org/10.1007/s12192-014-0564-x

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