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Image Search Results
Journal: Frontiers in Bioengineering and Biotechnology
Article Title: Improving the ex vivo expansion of human tumor-reactive CD8 + T cells by targeting toll-like receptors
doi: 10.3389/fbioe.2022.1027619
Figure Lengend Snippet: Co-stimulation with TLR1/2, TLR2/6, and TLR5 agonists enhances the proliferative response of human blood-derived CD8 + T cells to TCR triggering. (A) Expression profiles of surface TLRs in resting CD8 + T cells within human blood mononuclear cells (PBMCs). The T cell subpopulations of human PBMCs were identified by staining the cells with antibodies against CD3, CD4, and CD8; the TLR expression was measured by flow cytometry and expressed as the percentage of positively stained cells. Shown is aggregated data of 12 samples. (B and C) Co-stimulatory effects of different TLR agonists on the proliferation of human PBMC-derived CD8 + T cells. CD8+T cells were purified from human PBMCs, stained with eFluor 670, and then stimulated with ant-CD3 beads, anti-CD3/CD28 beads, or anti-CD3 beads coupled with the indicated TLRs agonists. Cells were harvested 7 days later and dilution of eFluor 670 was determined by flow cytometry to measure cellular proliferation, with representative flow cytometry plot and data summary (n = 3) shown in (B) and (C) , respectively. (D,E) Determination of optimal dosages of TLR agonists required for co-stimulation of ex vivo proliferation of human CD8 + T cells. CD8+T cells purified from human PBMCs were stimulated with anti-CD3 beads in presence of various concentration of TLR1/2 agonist, TLR2/6 agonist, or TLR5 agonist after labelling with eFluor 670. Flow cytometry analyses were performed on day 4 and 6 after stimulation to determine the cellular proliferation through assessment of eFluor 670 dilution. Shown is aggregated data of 3 samples.
Article Snippet: For surface staining, cells were stained in FACS buffer (PBS supplemented with 2% bovine serum albumin) sequentially with a fixable viability dye and a combination of the following antibodies: CD3-ECD (IM2705U, Beckman coulter), CD8-BV711(563677, BD Pharmingen), CD4-percp-cy5.5 (300530, Biolegend), CD14-BV786(563699, BD Pharmingen), TLR1-FITC(sc-47709FITC, Santa Cruz),
Techniques: Derivative Assay, Expressing, Staining, Flow Cytometry, Purification, Ex Vivo, Concentration Assay
Journal: Frontiers in Bioengineering and Biotechnology
Article Title: Improving the ex vivo expansion of human tumor-reactive CD8 + T cells by targeting toll-like receptors
doi: 10.3389/fbioe.2022.1027619
Figure Lengend Snippet: Co-stimulation with a combination of TLR1/2, TLR2/6, and TLR5 agonists enhances the transcriptional induction of proliferative-related genes in human blood-derived CD8 + T cells responding to TCR triggering. CD8+T cells purified from human PBMCs were stimulated with anti-CD3 beads in the presence or absence of a combination of TLR1/2 agonist, TLR2/6 agonist, and TLR5 agonist. Cells were harvested 24 h later for extracting total RNA, which was subjected to RNA sequencing. The differentially expressed genes showing >2-fold change between minus and plus TLR agonists groups were clustered based on Gene Ontology (GO) enrichment analysis (A) . Shown in (B) is the heatmap of representative proliferation-related genes that were identified to be co-stimulated by TLR agonists. H1 to H3 represent samples from three healthy individuals.
Article Snippet: For surface staining, cells were stained in FACS buffer (PBS supplemented with 2% bovine serum albumin) sequentially with a fixable viability dye and a combination of the following antibodies: CD3-ECD (IM2705U, Beckman coulter), CD8-BV711(563677, BD Pharmingen), CD4-percp-cy5.5 (300530, Biolegend), CD14-BV786(563699, BD Pharmingen), TLR1-FITC(sc-47709FITC, Santa Cruz),
Techniques: Derivative Assay, Purification, RNA Sequencing
Journal: Frontiers in Bioengineering and Biotechnology
Article Title: Improving the ex vivo expansion of human tumor-reactive CD8 + T cells by targeting toll-like receptors
doi: 10.3389/fbioe.2022.1027619
Figure Lengend Snippet: TLR agonists is an advantageous supplement benefiting the ex vivo expansion of human blood-derived PD1+ CD8 + T cells. (A) Exploration of optimal combination of TLR agonists in facilitating the ex vivo expansion of human blood-derived PD1+ CD8 + T cells. PD1+ CD8 + T cells isolated from blood samples of health individuals were continuously cultured for 21 days in medium containing anti-CD3/anti-CD28 beads and IL-7/IL-15, either alone or in the presence of different combination of TLR1/2, TLR2/6, and TLR5 agonists. Cell counting was performed at various time points throughout the culture period to generate the cell expansion curve. (B) Expansion curves of PD1+ CD8 + T cells derived from blood samples of cancer patients with different tumor types. Among the total of fifteen samples, seven were with lung cancer, four with pancreatic cancer, and the rest four with other types of cancers. Five samples from healthy individuals were also included as controls.
Article Snippet: For surface staining, cells were stained in FACS buffer (PBS supplemented with 2% bovine serum albumin) sequentially with a fixable viability dye and a combination of the following antibodies: CD3-ECD (IM2705U, Beckman coulter), CD8-BV711(563677, BD Pharmingen), CD4-percp-cy5.5 (300530, Biolegend), CD14-BV786(563699, BD Pharmingen), TLR1-FITC(sc-47709FITC, Santa Cruz),
Techniques: Ex Vivo, Derivative Assay, Isolation, Cell Culture, Cell Counting
Journal: PLOS ONE
Article Title: Toll-like receptors 1–9 in small bowel neuroendocrine tumors–Clinical significance and prognosis
doi: 10.1371/journal.pone.0302813
Figure Lengend Snippet: Baseline characteristics and comparison stratified by Toll-like receptor (TLR) 2 cytoplasmic intensity in primary tumors and lymph node metastases.
Article Snippet: After this, sections were incubated with rabbit polyclonal antibodies (TLRs 1, 2, 6, 7) and mouse monoclonal antibodies (TLRs 3, 4, 5, 8, 9) in dilute solution (Dako S2022); TLR1 for 60 minutes (diluted 1:300, Abcam ab189337),
Techniques: Comparison
Journal: PLOS ONE
Article Title: Toll-like receptors 1–9 in small bowel neuroendocrine tumors–Clinical significance and prognosis
doi: 10.1371/journal.pone.0302813
Figure Lengend Snippet: Immunohistochemical staining examples of Toll-like receptors (TLRs) 1, 2, 4, 5, 6, 7, 8 and 9 in representative small bowel neuroendocrine tumor samples showing ( A ) High TLR1 cytoplasmic intensity, ( B ) intermediate TLR2 cytoplasmic intensity, ( C ) intermediate TLR4 cytoplasmic intensity, ( D ) high TLR5 cytoplasmic and nucleic intensity, ( E ) Intermediate TLR6 cytoplasmic intensity, ( F ) intermediate TLR7 cytoplasmic intensity, ( G ) high TLR8 cytoplasmic intensity and ( H ) intermediate TLR9 cytoplasmic intensity in x20 magnification. The scale bar length is 50 μm (bottom left corner). Arrows indicate TLR-positive tumor cell islets.
Article Snippet: After this, sections were incubated with rabbit polyclonal antibodies (TLRs 1, 2, 6, 7) and mouse monoclonal antibodies (TLRs 3, 4, 5, 8, 9) in dilute solution (Dako S2022); TLR1 for 60 minutes (diluted 1:300, Abcam ab189337),
Techniques: Immunohistochemical staining, Staining
Journal: PLOS ONE
Article Title: Toll-like receptors 1–9 in small bowel neuroendocrine tumors–Clinical significance and prognosis
doi: 10.1371/journal.pone.0302813
Figure Lengend Snippet: Median Toll-like receptor (TLR) staining intensity in primary small bowel neuroendocrine tumors and lymph node metastases.
Article Snippet: After this, sections were incubated with rabbit polyclonal antibodies (TLRs 1, 2, 6, 7) and mouse monoclonal antibodies (TLRs 3, 4, 5, 8, 9) in dilute solution (Dako S2022); TLR1 for 60 minutes (diluted 1:300, Abcam ab189337),
Techniques: Staining
Journal: PLOS ONE
Article Title: Toll-like receptors 1–9 in small bowel neuroendocrine tumors–Clinical significance and prognosis
doi: 10.1371/journal.pone.0302813
Figure Lengend Snippet: Disease-specific survival rates based on Toll-like receptor (TLR) 1, 2, 4, 5, 6, 7, 8, and 9 cytoplasmic staining intensity and TLR5 nucleic staining intensity in both primary tumors and lymph node metastases.
Article Snippet: After this, sections were incubated with rabbit polyclonal antibodies (TLRs 1, 2, 6, 7) and mouse monoclonal antibodies (TLRs 3, 4, 5, 8, 9) in dilute solution (Dako S2022); TLR1 for 60 minutes (diluted 1:300, Abcam ab189337),
Techniques: Staining
Journal: Journal of Virology
Article Title: Early Innate Recognition of Herpes Simplex Virus in Human Primary Macrophages Is Mediated via the MDA5/MAVS-Dependent and MDA5/MAVS/RNA Polymerase III-Independent Pathways
doi: 10.1128/JVI.01106-10
Figure Lengend Snippet: HSV and PAMPs induce IFN, ISGs, and proinflammatory cytokines in human primary macrophages. (A to C and G) Macrophages were infected with HSV-1 (17+, 5 × 105 PFU/ml, MOI of 1) or transfected with poly(I:C) [t-poly(I:C); 2 μg/ml] or poly(dA:dT) [t-poly(dA:dT); 2 μg/ml]. (A to C and G) After 6 h, after which RNA was harvested and mRNA accumulation of IFN-β, IFN-λ1, CXCL10, and TNF-α was assayed by real-time PCR. (D, E, and H) In similar experiments, cell supernatants were harvested after 20 h and assayed for the presence of TNF-α, CXCL10, and CCL3 by ELISA. (E, F, I, and K) Macrophages were infected with HSV or stimulated with IFN-α (100 IU/ml) or the agonist for TLR2 (Pam3CSK4, 100 ng/ml), TLR3 [poly(I:C), 30 μg/ml], TLR7/8 (R848, 1 μg/ml), or TLR9 (ODN2216, 2 μg/ml). After 20 h of stimulation, supernatants were collected and protein levels assayed by ELISA. (J and L) Cells were infected with HSV-1 (17+ or KOS, 5 × 105 PFU/ml, MOI of 1) or HSV-2 (MOI of 1). Supernatants were harvested at 18 h and assayed by ELISA for CXCL10 and CCL3. The real-time PCR data are depicted as relative units (RU), which is a fold change in gene expression normalized to that of the endogenous reference gene 18S rRNA and is relative to the no template control (NTC) calibrator. The results depicted are means ± standard deviations (SD) within the shown experiment. Experiments were done five times (A and B) or two times (C to L), with similar findings.
Article Snippet:
Techniques: Infection, Transfection, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing
Journal: Journal of Virology
Article Title: Early Innate Recognition of Herpes Simplex Virus in Human Primary Macrophages Is Mediated via the MDA5/MAVS-Dependent and MDA5/MAVS/RNA Polymerase III-Independent Pathways
doi: 10.1128/JVI.01106-10
Figure Lengend Snippet: HSV-1 infection activates cytokine expression independently of TLR2. Cells were infected with HSV-1 (17+, MOI of 1) or stimulated with TLR2 ligand Pam3CSK4 (100 ng/ml) in the presence or absence of TLR2 neutralizing antibodies (8.5 μg/ml). After 6 h, RNA was harvested, and after 18 h, supernatants were collected. Accumulation of TNF-α and IFN-β mRNA was assessed by real-time PCR (B and C), and the amounts of secreted TNF-α, CXCL10, and CCL3 were measured by ELISA (A, D, and E). RU, relative units. The results depicted are means ± SD from one of two experiments showing similar results.
Article Snippet:
Techniques: Infection, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay