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

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.

Peste des Petits Ruminants Virus Enters Caprine Endometrial Epithelial Cells via the Caveolae-Mediated Endocytosis Pathway

“…Endometrial epithelial cell were immortalized by transfection with human telomerase reverse transcriptase, and we confirmed that their secretory function was consistent with that of primary EECs ( Qi X. et al, 2012 ; Qi X.F. et al, 2012 ; Wang et al, 2015 ) kindly provided by Prof. Yaping Jin (Northwest A&F University Yangling, Shaanxi, China). As a result of immortalization, a portion of the EECs were prone to aging and became larger and longer than primary cells with increased passage.…”

Hormone regulates endometrial function via cooperation of endoplasmic reticulum stress and mTOR‐autophagy

“…Goat endometrial epithelial cells (EECs) and goat trophoblast cells (GTCs) were immortalized by transfection with human telomerase reverse transcriptase (hTERT), and were well stored in our laboratory (Dong et al, ; Wang et al, ; Zhang, Wang, Wu, Sheng, & Jin, ). The EECs and GTCs were seeded in six‐well plates cultured in DMEM/F‐12 medium containing 10% FBS (Cornig, Manassas, VA), and incubated at 37°C in humidified 5% CO 2 incubator.…”

Palmitic acid induces human osteoblast-like Saos-2 cell apoptosis via endoplasmic reticulum stress and autophagy

“…The apoptosis level of Saos-2 cells was quantified with an Annexin V/PI Apoptosis Analysis Kit according to the manufacturer's instructions. The cells were analyzed using a fluorescence-activated cell sorter (Becton, Dickinson and Company, USA) within 1 h. Early apoptotic cells were determined by counting the percentage of Annexin V+/PI− cells; progressed apoptotic cells were obtained by counting the percentage of Annexin V+/PI+ cells; necrotic cells were detected by counting the percentage of Annexin V-/PI+ cells, and Annexin V−/PI− cells were considered as surviving cells as the previous reported (Lin et al 2015;Lin et al 2012;Wang et al 2016;Wang et al 2015).…”