Innate immune transcriptomic evaluation of PBMC isolated from sheep after infection with E. ruminantium Welgevonden strain

Nefefe, Tshifhiwa; Liebenberg, Junita; Kleef, Mirinda Van; Steyn, H.C.; Pretorius, Ashley

doi:10.1016/j.molimm.2017.09.018

Cited by 10 publications

(3 citation statements)

References 68 publications

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“…About 10 mL of blood was collected from each of the three healthy sheep, and PBMCs were separately isolated by polysucrose-diammonia density gradient centrifugation according to the instructions of the manufacturer (TMD Science, China) (36). The three PBMC samples were washed in three volumes of pre-chilled PBS, and cell viability was checked using trypan blue.…”

Section: Isolation Of Sheep Pbmcsmentioning

confidence: 99%

Identification of Different Extracellular Vesicles in the Hydatid Fluid of Echinococcus granulosus and Immunomodulatory Effects of 110 K EVs on Sheep PBMCs

Yang

et al. 2021

Front. Immunol.

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Echinococcosis, mainly caused by Echinococcus granulosus, is one of the 17 neglected tropical diseases. Extracellular vesicles (EVs) play an essential role in the host–parasite interplay. However, the EVs in the hydatid fluid (HF) of E. granulosus are not fully characterized. Herein, three different types of HF EVs, designated as 2 K, 10 K, and 110 K EVs based on the centrifugal force used, were morphologically identified. A total of 97, 80, and 581 proteins were identified in 2 K, 10 K, and 110 K EVs, respectively, 39 of which were commonly shared. Moreover, 11, 8, and 25 miRNAs were detected, respectively, and all of the 7 selected miRNAs were validated by qPCR to be significantly lower abundant than that in protoscoleces. It was further deemed that 110 K EVs were internalized by sheep peripheral blood mononuclear cells (PBMCs) in a time-dependent manner and thus induced interleukin (IL)-10, tumor necrosis factor-α (TNF-α), and IRF5 were significantly upregulated and IL-1β, IL-17, and CD14 were significantly downregulated (p < 0.05). These data demonstrate the physical discrepancy of three HF EVs and an immunomodulatory effect of 110 K EVs on sheep PMBCs, suggesting a role in immune responses during E. granulosus infection.

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Section: Isolation Of Sheep Pbmcsmentioning

confidence: 99%

Identification of Different Extracellular Vesicles in the Hydatid Fluid of Echinococcus granulosus and Immunomodulatory Effects of 110 K EVs on Sheep PBMCs

Yang

et al. 2021

Front. Immunol.

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“…Peripheral blood mononuclear cells (PBMCs) were isolated from three healthy sheep by polysucrose-diammonia density gradient centrifugation, according to the instructions of the manufacturer (TMD Science) [ 25 ]. The three PBMC samples were washed in 3× volumes of pre-chilled phosphate-buffered saline (PBS) before inoculation into cell culture flasks, followed by checking their viability by Trypan blue exclusion (Sigma-Aldrich, St. Louis, MO, USA).…”

Section: Methodsmentioning

confidence: 99%

Comparative characterization of microRNA-71 of Echinococcus granulosus exosomes

Yan,

Li,

et al. 2023

Parasite

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Cystic echinococcosis (CE) is a global zoonotic disease caused by Echinococcus granulosus, posing a great threat to human and animal health. MiRNAs are small regulatory noncoding RNA involved in the pathogenesis of parasitic diseases, possibly via exosomes. Egr-miR-71 has been identified as one of the miRNAs in the blood of CE patients, but its secretory characteristics and functions remains unclear. Herein, we studied the secretory and biological activity of exosomal egr-miR-71 and its immunoregulatory functions in sheep peripheral blood mononuclear cells (PBMCs). Our results showed that egr-miR-71 was enriched in the exosome secreted by protoscoleces with biological activity. These egr-miR-71-containing exosomes were easily internalized and then induced the dysregulation of cytokines (IL-10 and TNF-α), nitric oxide (NO) and key components (CD14 and IRF5) in the LPS/TLR4 pathway in the coincubated sheep PBMCs. Similarly, egr-miR-71 overexpression also altered the immune functions but exhibited obvious differences in regulation of the cytokines and key components, preferably inhibiting proinflammatory cytokines (IL-1α, IL-1β and TNF-α). These results demonstrate that exosomal egr-miR-71 is bioactive and capacity of immunomodulation of PBMCs, potentially being involved in immune responses during E. granulosus infection.

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“…Several in vitro and in vivo studies were conducted on different species to understand the role of LPS and LTA. These molecules are effective on activating innate and adaptive immune system and the function of TLRs in these immune systems (Carroll et al 2012;Deng et al 2012;Gao et al 2010;Islam et al 2012;Nefefe et al 2017;Passmore et al 2016;Xu et al 2017). The innate immune system is known to be rapidly activated within 0-4 h to eliminate foreign molecules in the body, give early stimulated response at the 4-96 h (innate-adaptive immunity), and transfer the task to the adaptive immune cells after the 96 h (Murphy 2014).…”

Section: Introductionmentioning

confidence: 99%

Effect of LPS and LTA stimulation on the expression of TLR-pathway genes in PBMCs of Akkaraman lambs in vivo

Aksel

Akyüz

2021

Trop Anim Health Prod

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This is the first study investigating the changes in some gene expressions related to the TLR pathway in vivo in sheep. Lipopolysaccharide (LPS) and lipoteichoic acid (LTA) molecules were administrated separately and in combination to the Akkaraman lambs via intranasal route. For this purpose, 28 lambs were distributed into four groups (LPS, LTA, LPS + LTA, and control, n = 7). Blood samples were collected to isolate the peripheral blood mononuclear cells (PBMCs) at 24 h and on day 7. Expression levels of TLR2, TLR4, MyD88, TRAF6, TNF-α, IL-1ß, IL-6, IL-10, NF-κß, and IFN-γ genes were determined by qRT-PCR. Increases were determined in the expression data of TLR2 [LPS (P < 0.05) and LTA + LPS (P < 0.01)], TLR4 [LTA + LPS (P < 0.05)], TNF-α, IL-10 [LTA + LPS (P < 0.05)], and IFN-γ genes in all groups in the mRNA expression analysis of PBMCs isolated at 24 h whereas decreases were determined in the expression levels of these genes on day 7. The combination of LPS + LTA stimulated lamb PBMCs more effectively than separate administration of LPS and LTA at 24 h. Therefore, this article may contribute to the understanding the host-pathogen interaction of respiratory-transmitted bacterial diseases concerning PBMCs at 24 h and on day 7. Also this study may contribute to the dose adjustment for bacterial vaccine studies in sheep. Experimental application doses will be helpful for in vivo and in vitro drug and vaccine development studies in the fields of pharmacology and microbiology.

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Innate immune transcriptomic evaluation of PBMC isolated from sheep after infection with E. ruminantium Welgevonden strain

Cited by 10 publications

References 68 publications

Identification of Different Extracellular Vesicles in the Hydatid Fluid of Echinococcus granulosus and Immunomodulatory Effects of 110 K EVs on Sheep PBMCs

Identification of Different Extracellular Vesicles in the Hydatid Fluid of Echinococcus granulosus and Immunomodulatory Effects of 110 K EVs on Sheep PBMCs

Comparative characterization of microRNA-71 of Echinococcus granulosus exosomes

Effect of LPS and LTA stimulation on the expression of TLR-pathway genes in PBMCs of Akkaraman lambs in vivo

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