Journal: Virology Journal

Article Title: Detection of hepatitis C virus (HCV) negative strand RNA and NS3 protein in peripheral blood mononuclear cells (PBMC): CD3 + , CD14 + and CD19 +

doi: 10.1186/1743-422X-10-346

Figure Lengend Snippet: Demographic, clinical and laboratory data of the patients

Article Snippet: For separation of particular cell subpopulations, the following antibodies were used: anti-CD3 (anti-human CD3 magnetic Particles-DM, BD IMag™, San Jose, USA), anti-CD19 (anti-human CD19 magnetic Particles-DM, BD IMag™; San Jose, USA), anti-CD14 (anti-human CD14 magnetic Particles-DM, BD IMag™; San Jose, USA).

Techniques:

Journal: Virology Journal

Article Title: Detection of hepatitis C virus (HCV) negative strand RNA and NS3 protein in peripheral blood mononuclear cells (PBMC): CD3 + , CD14 + and CD19 +

doi: 10.1186/1743-422X-10-346

Figure Lengend Snippet: Detection of NS3 HCV in PBMC subpopulations: CD3 + , CD14 + and CD19 + (Pt 1). Negative control CD3 + , CD14 + and CD19 + subpopulations were collected from HCV - negative patient. Liver tissue control: liver biopsy samples from HCV positive patient (HCV pos.) and HCV negative patient (HCV neg.).

Article Snippet: For separation of particular cell subpopulations, the following antibodies were used: anti-CD3 (anti-human CD3 magnetic Particles-DM, BD IMag™, San Jose, USA), anti-CD19 (anti-human CD19 magnetic Particles-DM, BD IMag™; San Jose, USA), anti-CD14 (anti-human CD14 magnetic Particles-DM, BD IMag™; San Jose, USA).

Techniques: Negative Control

Journal: Virology Journal

Article Title: Detection of hepatitis C virus (HCV) negative strand RNA and NS3 protein in peripheral blood mononuclear cells (PBMC): CD3 + , CD14 + and CD19 +

doi: 10.1186/1743-422X-10-346

Figure Lengend Snippet: The purity of CD3 + (A), CD14 + (B) and CD19 + (C) cell subpopulations separated from PBMC collected from HCV positive patient.

Article Snippet: For separation of particular cell subpopulations, the following antibodies were used: anti-CD3 (anti-human CD3 magnetic Particles-DM, BD IMag™, San Jose, USA), anti-CD19 (anti-human CD19 magnetic Particles-DM, BD IMag™; San Jose, USA), anti-CD14 (anti-human CD14 magnetic Particles-DM, BD IMag™; San Jose, USA).

Techniques:

Journal: Journal of translational medicine

Article Title: Intrahepatic macrophage reprogramming associated with lipid metabolism in hepatitis B virus-related acute-on-chronic liver failure.

doi: 10.1186/s12967-023-04294-1

Figure Lengend Snippet: Fig. 6 The targeted lipid metabolomics in the livers of cirrhosis and ACLF patients and stimulation test with α-LA. a The volcano plot of FFAs in the livers showed as the ACLF group over the cirrhosis group. 23 FFAs were increased in ACLF group while only 1 FFA was increased in cirrhosis group. b The 24 differentiated FFAs for each sample shown by Z score. c The metabolite sets enrichment analysis for the increased FFAs in ACLF livers. d The expression of TREM2 on selected CD14+ peripheral monocytes stimulated in vitro. The expression of TREM2 was significantly increased when stimulated with α-LA (60 μM) and LPS (100 ng/mL). (**p < 0.01, ****p < 0.0001, tested by paired t-test)

Article Snippet: Human peripheral CD14+ monocytes were positively selected from PBMCs of eight HCs using magnetic activated cell sorting (MACS) CD14 microbeads (Miltenyi Biotec, 130-050-201).

Techniques: Expressing, In Vitro

Journal: The Journal of Biological Chemistry

Article Title: In silico interrogation of the miRNAome of infected hematopoietic cells to predict processes important for human cytomegalovirus latent infection

doi: 10.1016/j.jbc.2023.104727

Figure Lengend Snippet: HCMV infection of myeloid cells triggers induction of host miRNAs. A , CD34+ cells were infected at MOI = 5 with HCMV strain Merlin (n = 3). Media was replaced 3 h post-infection, and 6 hours post-infection; miRNA was extracted and assessed by qPCR array for 1055 miRNAs. Orange triangles represent miRNAs upregulated >2-fold with P < 0.05, blue diamonds represent miRNAs downregulated >2-fold with P < 0.05, P -values calculated by student’s t test. B , Primary CD34+ cells were infected at MOI = 5 with Merlin or TB40/E (n = 2 independent replicates). Six hours post-infection, miRNA was extracted and assessed by specific qRT-PCR, with expression displayed as −ΔCt relative to expression in uninfected cells or a technical cutoff. C and D , Primary CD14+ monocytes ( C ) or primary fibroblasts ( D ) were infected at MOI = 5 with Merlin (n = 2 independent replicates). Six hours post-infection, miRNA was extracted and assessed by specific qRT-PCR, with expression displayed as −ΔCt relative to expression in uninfected cells or a technical cutoff if undetected in uninfected cells (indicated by #). All summary values are displayed as mean±SD. HCMV, Human cytomegalovirus; miRNAs, microRNA.

Article Snippet: CD14+ cells were obtained from healthy volunteers (NHS London Hampstead research ethic committee - 08/H0720/46) by initial centrifugation of PBS-diluted whole blood over Histopaque-1077 to isolate PBMCs, followed by positive magnetic selection (CD14+ microbeads, MS columns, Miltenyi Biotec).

Techniques: Infection, Quantitative RT-PCR, Expressing

Journal: The Journal of Biological Chemistry

Article Title: In silico interrogation of the miRNAome of infected hematopoietic cells to predict processes important for human cytomegalovirus latent infection

doi: 10.1016/j.jbc.2023.104727

Figure Lengend Snippet: Calcium channel inhibitors prevent expression from the MIEP selectively during reactivation. A , THP-1 cells were infected at MOI = 5 with HCMV strain Merlin. At 1hpi, cells were treated with calcium channel inhibitors or solvent control. At 24hpi, RNA was harvested, and total IE gene expression was quantified by qRT-PCR (n = 2 independent replicates). B , Primary CD14+ monocytes were infected at MOI = 5 with HCMV strain Merlin. After 3 days, differentiation into MoDCs was initiated with IL-4 and GM-CSF. Six days later, cells were treated with calcium channel inhibitors for 3 h prior to IL-6 treatment, and viral IE gene expression was quantified by qRT-PCR 24h later (n = 3 independent replicates). Summary data shown as mean±SD. Statistical comparison by one-way ANOVA with non-parametric Kruskal–Wallis test with Dunn’s multiple comparison correction. GM-CSF, granulocyte/macrophage-colony stimulating factor; HCMV, Human cytomegalovirus.

Article Snippet: CD14+ cells were obtained from healthy volunteers (NHS London Hampstead research ethic committee - 08/H0720/46) by initial centrifugation of PBS-diluted whole blood over Histopaque-1077 to isolate PBMCs, followed by positive magnetic selection (CD14+ microbeads, MS columns, Miltenyi Biotec).

Techniques: Expressing, Infection, Quantitative RT-PCR