trem 1  (Santa Cruz Biotechnology)

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: ox-LDL-induced DCs TREM-1 expression. (a) DCs at day 6 were treated with LDL or ox-LDL at different time (0, 6, 12, and 24 h) or at different concentration (25, 50, or 100 μ g mL −1 ) for 24 h. Then, the protein levels of TREM-1 were determined. (b) DCs at day 6 were treated with LDL or ox-LDL at 50 μ g mL −1 at 0, 6, 12, and 24 h, and then the mRNA levels of TREM-1 were determined. (c) DCs at day 6 were treated with ox-LDL at 25, 50, or 100 μ g mL −1 for 24 h, and then the mRNA levels of TREM-1 were determined. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL-treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Expressing, Concentration Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: ox-LDL-induced DCs TREM-1 expression with or without LP17 and SiTREM-1. At day 6, DCs were treated with LDL or ox-LDL for 24h; 100 ng/mL LPS treatment is a positive control. (a) Cell-free supernatants were harvested and assayed for sTREM-1 content by ELISA (b) TREM-1 surface expression. DCs were stained with anti-CD1a-allophycocyanin and anti-TREM-1-PE Abs and analyzed by flow cytometry. Cells were electronically gated according to their light scatter properties to exclude cell debris. Data are expressed as a percentage of TREM-1 + cells within CD1a + DCs and ox-LDL-DCs generated from healthy volunteer donors. (c, d) For downregulation of TREM-1, DCs was transfected with TREM-1 silencer (SiTREM-1) or treated with LP17 at day 6, and ox-LDL was added 24 h after transfection and further incubated for 24 h. Then, the mRNA levels and protein levels of TREM-1 were determined. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Expressing, Positive Control, Enzyme-linked Immunosorbent Assay, Staining, Flow Cytometry, Generated, Transfection, Incubation

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: TREM-1 plays an important role in DCs maturation and subsequent T-cell proliferation induced by ox-LDL. (a) The process of monocytes differentiated into immature DCs (imDC), at 0, 1, 2, 3, 4, 5, and 6 days; the expressions of CD14 were determined by flow cytometry. (b) At day 6, immature DCs were treated with LDL or ox-LDL for 24 h, and the expressions of CD1a, CD40, CD86, CD83, and HLA-DR on DCs were analyzed by flow cytometry. 100 ng/mL LPS treatment is a positive control. (c, d) CXCR4 and CCCR7 on DCs were analyzed by flow cytometry. 100 ng/mL LPS treatment is a positive control. (e) T- cell proliferation determined by BrdU incorporation assay. After the indicated treatment, DCs at day 7 were washed and cocultured in triplicate for 5 days with autologous T cells. 10 μ mol L−1 BrdU was present in the last 16 h. Proliferative response was evaluated as OD value. Results represent the mean ± SD of seven-eight experiments. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Flow Cytometry, Positive Control, BrdU Incorporation Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: TREM-1 regulated ox-LDL-induced DCs inflammation. (a–f) At day 6, DCs were transfected with or without TREM-1 silencer (SiTREM-1) or relevant scrambled siRNA or treated with LP17, and ox-LDL was added 24 h after transfection and further incubated for 24 h. DC supernatants were collected, and cytokine production were tested by ELISA. (g, h) At day 6, DCs were treated with ox-LDL at different concentrations (25, 50, or 100 μ g mL − 1 ) for 24 h, IL-12 was determined by ELISA, and 100 ng/mL LPS treatment is a positive control. Results represent the mean ± SD of seven-eight experiments. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL-treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Transfection, Incubation, Enzyme-linked Immunosorbent Assay, Positive Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: T-cell polarization-related transcription factors, TBX21, GATA3, RORC, and FOXP3 gene expressions, were evaluated by real-time PCR. ox-LDL-treated DCs strongly induced TBX21 and/or RORC expression in naïve T-cell populations. However, TREM-1 inhibition suppressed the expression of TBX21 and RORC while inducing the expression of FoxP3. Results represent the mean ± SD of seven experiments. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL-treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Inhibition

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: TREM-1 mediated the Th1/Th17-priming induced by ox-LDL-treated DCs. (a–e) IFN- γ , IL- 17, IL- 4, TGF- β , and IL- 10 content were determined by ELISA. (f) T [DC (siR-TREM-1 + ox-LDL)] cells suppressed primary T-cell proliferation. Naïve CD4 + T cells were cultured with ox-LDL- or SiTREM-1 + ox-LDL-treated autologous DCs for 13 days. Naïve CD4 + T cells labeled with 5 μ mol L −1 CFSES were stimulated with mature DCs (mDCs) alone or in the presence of T [DC], T [DC (ox-LDL)], T [DC (Si-TREM-1 + ox-LDL)], or T [DC (Mock + ox-LDL)] cells at a 1 : 1 ratio. T-cell proliferation was measured by CFSE dilution. Results represent the mean ± SD of seven experiments. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL-treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Labeling

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: miRNA-155/SOCS1 signaling pathways were involved in the effect of TREM-1 and ox-LDL. (a) Expression of miRNAs in DCs was tested by RT-PCR. The value was normalized as a fold change to that of nontreated DC samples. (b-c) DCs were transfected with miRNA-155 inhibitor or mimic, respectively, and stimulated with ox-LDL. SOCS1 level in DCs were detected by Western blot (b) and RT-PCR (c). Results represent the mean ± SD of seven experiments. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL-treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: TREM-1 expression in DCs from symptomatic and asymptomatic patients with carotid stenosis and SOCS1 signaling pathways were involved in the effect of TREM-1 and ox-LDL on DCs from patients with carotid stenosis. (a) The mRNA and protein of TREM-1 expression in DCs from symptomatic were higher than from asymptomatic patients with carotid stenosis; (b) DCs were generated from peripheral blood monocytes of patients who underwent carotid endarterectomy. miR-155 and SOCS1 mRNA expression were determined by RT-PCR. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL-treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Expressing, Generated, Reverse Transcription Polymerase Chain Reaction

Journal: Oxidative Medicine and Cellular Longevity

Article Title: TREM-1 Modulates Dendritic Cells Maturation and Dendritic Cell-Mediated T-Cell Activation Induced by ox-LDL

doi: 10.1155/2022/3951686

Figure Lengend Snippet: TREM-1 was involved in DC maturation, proinflammatory cytokine production, and T-cell proliferation in plaque patients induced by ox-LDL. DCs were generated from peripheral blood monocytes of patients who underwent carotid endarterectomy. The same protocol is used as that for DCs of normal donors. Briefly, DCs at day 6 were treated with 50 μ g mL − 1 ox-LDL for 24 h. (a) The expressions of CD1a, CD40, CD86, CD83, and HLA‐DR on day 7 DCs were analyzed by flow cytometry. (b) T‐cell proliferation determined by BrdU incorporation assay. After the indicated treatment, DCs were washed and cocultured in triplicate for 5 days with autologous T cells. 10 μ mol L − 1 BrdU was present in the last 16 h. Proliferative response was evaluated as OD value. (c) Cytokines production were tested by ELISA. Results represent the mean ± SD of seven-eight experiments. ∗ p < 0.05, compared with control cells; # p < 0.05, compared with ox-LDL-treated cells.

Article Snippet: Lipofectamine® 2000 Transfection Reagent and TREM-1 siRNA were purchased from Invitrogen (Carlsbad, USA).

Techniques: Generated, Flow Cytometry, BrdU Incorporation Assay, Enzyme-linked Immunosorbent Assay