Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Impact of patient characteristics on innate immune responses and inflammasome activation in ex vivo human lung tissues infected with influenza A virus

doi: 10.3389/fcimb.2023.1269329

Figure Lengend Snippet: Difference in the expressions of molecular biomarkers in immune cells and secreted cytokines in culture supernatants between mock infection and influenza A(H3N2) infection groups after 24 h post-infection.

Article Snippet: For TLR detection , BV421 mouse anti-human CD281 (TLR1) antibody (catalog no. 566430, Becton Dickinson Biosciences, San Jose, CA, USA) and BV510 mouse anti-human CD282 (TLR2) (catalog no. 742767, Becton Dickinson Biosciences, San Jose, CA, USA), and PE mouse anti-human CD283 (TLR3) antibody (catalog no. 315009, BioLegend, Inc., San Diego, CA, USA) were used.

Techniques: Infection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Impact of patient characteristics on innate immune responses and inflammasome activation in ex vivo human lung tissues infected with influenza A virus

doi: 10.3389/fcimb.2023.1269329

Figure Lengend Snippet: Associations of % differences in nucleoprotein (NP), toll-like receptors (TLRs) and programmed death 1 (PD-1)/programmed death-ligand 1 (PD-L1) pathways with % differences in secreted cytokine levels.

Article Snippet: For TLR detection , BV421 mouse anti-human CD281 (TLR1) antibody (catalog no. 566430, Becton Dickinson Biosciences, San Jose, CA, USA) and BV510 mouse anti-human CD282 (TLR2) (catalog no. 742767, Becton Dickinson Biosciences, San Jose, CA, USA), and PE mouse anti-human CD283 (TLR3) antibody (catalog no. 315009, BioLegend, Inc., San Diego, CA, USA) were used.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Impact of patient characteristics on innate immune responses and inflammasome activation in ex vivo human lung tissues infected with influenza A virus

doi: 10.3389/fcimb.2023.1269329

Figure Lengend Snippet: Comparison of % differences in the variables of interest between the age ≥ 65 years group and age < 65 years group.

Article Snippet: For TLR detection , BV421 mouse anti-human CD281 (TLR1) antibody (catalog no. 566430, Becton Dickinson Biosciences, San Jose, CA, USA) and BV510 mouse anti-human CD282 (TLR2) (catalog no. 742767, Becton Dickinson Biosciences, San Jose, CA, USA), and PE mouse anti-human CD283 (TLR3) antibody (catalog no. 315009, BioLegend, Inc., San Diego, CA, USA) were used.

Techniques: Comparison

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Impact of patient characteristics on innate immune responses and inflammasome activation in ex vivo human lung tissues infected with influenza A virus

doi: 10.3389/fcimb.2023.1269329

Figure Lengend Snippet: Differential expressions of NP (A) , TLR1 (B) , TLR2 (C) , and TLR3 (D) on macrophages, TNF-α levels in culture supernatants (E) , IL-2 levels in culture supernatants (F) , and IFN-γ levels in culture supernatants (G) across Cluster 1, Cluster 2, and Cluster 3. The presented data are represented as medians with interquartile ranges. Between-group comparisons were conducted using the independent-samples Kruskal-Wallis test with Bonferroni correction. IFN-γ, interferon-gamma; IL, interleukin; NP, nucleoprotein; TLR, toll-like receptor; TNF-α, tumor necrosis factor-alpha.

Article Snippet: For TLR detection , BV421 mouse anti-human CD281 (TLR1) antibody (catalog no. 566430, Becton Dickinson Biosciences, San Jose, CA, USA) and BV510 mouse anti-human CD282 (TLR2) (catalog no. 742767, Becton Dickinson Biosciences, San Jose, CA, USA), and PE mouse anti-human CD283 (TLR3) antibody (catalog no. 315009, BioLegend, Inc., San Diego, CA, USA) were used.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Impact of patient characteristics on innate immune responses and inflammasome activation in ex vivo human lung tissues infected with influenza A virus

doi: 10.3389/fcimb.2023.1269329

Figure Lengend Snippet: Comparisons of clinical characteristics and variables of interest after mock infection across the three clusters.

Article Snippet: For TLR detection , BV421 mouse anti-human CD281 (TLR1) antibody (catalog no. 566430, Becton Dickinson Biosciences, San Jose, CA, USA) and BV510 mouse anti-human CD282 (TLR2) (catalog no. 742767, Becton Dickinson Biosciences, San Jose, CA, USA), and PE mouse anti-human CD283 (TLR3) antibody (catalog no. 315009, BioLegend, Inc., San Diego, CA, USA) were used.

Techniques: Infection, Expressing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Impact of patient characteristics on innate immune responses and inflammasome activation in ex vivo human lung tissues infected with influenza A virus

doi: 10.3389/fcimb.2023.1269329

Figure Lengend Snippet: Comparative analysis of the % differences in the variables of interest among the three clusters.

Article Snippet: For TLR detection , BV421 mouse anti-human CD281 (TLR1) antibody (catalog no. 566430, Becton Dickinson Biosciences, San Jose, CA, USA) and BV510 mouse anti-human CD282 (TLR2) (catalog no. 742767, Becton Dickinson Biosciences, San Jose, CA, USA), and PE mouse anti-human CD283 (TLR3) antibody (catalog no. 315009, BioLegend, Inc., San Diego, CA, USA) were used.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Mycobacterial Phosphatidylinositol Mannoside 6 (PIM 6 ) Up-Regulates TCR-Triggered HIV-1 Replication in CD4 + T Cells

doi: 10.1371/journal.pone.0080938

Figure Lengend Snippet: (A) TLR2 protein expression on JLTRg-R5 cells before (JLTRg-R5) and after transfection with empty vector (JLTRg-R5-empty) or with vector encoding TLR2/1 (JLTRg-R5-TLR1-TLR2) was determined by flow cytometry. (B) Expression of TLR2 mRNA in JLTRg-R5 before (JLTRg-R5) and after transfection with empty vector (JLTRg-R5-empty) or with vector encoding TLR2/1 (JLTRg-R5-TLR1-TLR2) was measured by RT-PCR. Primary monocytes (MN) served as positive control. Shown is one representative experiment of three.

Article Snippet: The transfection efficiency was determined by flow cytometry using anti-human CD282 (TLR2, clone 2.5) and anti-human CD281 (TLR1) both from e-Biosciences and confirmed by RNA expression by RT-PCR using TLR1 and TLR2 primers from Invivogen.

Techniques: Expressing, Transfection, Plasmid Preparation, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Positive Control

Journal: PLoS ONE

Article Title: Mycobacterial Phosphatidylinositol Mannoside 6 (PIM 6 ) Up-Regulates TCR-Triggered HIV-1 Replication in CD4 + T Cells

doi: 10.1371/journal.pone.0080938

Figure Lengend Snippet: JLTRg-R5-TLR1-TLR2 and JLTRg-R5-empty were activated overnight with α-CD3 in medium alone (None) or medium containing LprG (LprG) or PIM 6 (PIM 6 ). Cells were then infected with NLAD8 virus for 72 h and viral replication determined by measuring intracellular p24 by flow cytometry ( A ) or RT activity in culture supernatants ( B ). (A) Shown are means ± SD of four independent experiments. (B) Shown are means ± SD of two independent experiments in triplicate.

Article Snippet: The transfection efficiency was determined by flow cytometry using anti-human CD282 (TLR2, clone 2.5) and anti-human CD281 (TLR1) both from e-Biosciences and confirmed by RNA expression by RT-PCR using TLR1 and TLR2 primers from Invivogen.

Techniques: Infection, Flow Cytometry, Activity Assay

Journal: Journal of Immunology Research

Article Title: Analysis of Toll-Like Receptors in Human Milk: Detection of Membrane-Bound and Soluble Forms

doi: 10.1155/2019/4078671

Figure Lengend Snippet: List of antibodies used in this study.

Article Snippet: Anti-TLR1 , Polyclonal rabbit antibody, directed against a recombinant peptide from human TLR1 (STJ25862) , St John's Laboratory , 1 : 500.

Techniques: Recombinant

Journal: Journal of Immunology Research

Article Title: Analysis of Toll-Like Receptors in Human Milk: Detection of Membrane-Bound and Soluble Forms

doi: 10.1155/2019/4078671

Figure Lengend Snippet: Sequences of TLR1, TLR2, TLR4, and TLR6 mature forms, preceded by UniProt identifier, initial and final amino acid number, and estimated molecular weight.

Article Snippet: Anti-TLR1 , Polyclonal rabbit antibody, directed against a recombinant peptide from human TLR1 (STJ25862) , St John's Laboratory , 1 : 500.

Techniques: Molecular Weight

Journal: Journal of Immunology Research

Article Title: Analysis of Toll-Like Receptors in Human Milk: Detection of Membrane-Bound and Soluble Forms

doi: 10.1155/2019/4078671

Figure Lengend Snippet: Immunoblot of TLR1/2/4/6 in MFGMs of colostrum. Gestational age is indicated as “T” (term) or “PT” (preterm). Each number refers to a different milk sample. Standard molecular weights are indicated on the left. Each box shows the blot reactivity to a specific anti-TLR antibody and, on the lower side, the same blot incubated with an anti- β -actin antibody.

Article Snippet: Anti-TLR1 , Polyclonal rabbit antibody, directed against a recombinant peptide from human TLR1 (STJ25862) , St John's Laboratory , 1 : 500.

Techniques: Western Blot, Incubation

Journal: Journal of Immunology Research

Article Title: Analysis of Toll-Like Receptors in Human Milk: Detection of Membrane-Bound and Soluble Forms

doi: 10.1155/2019/4078671

Figure Lengend Snippet: Immunoblot of TLR1/2/4/6 in MFGMs of mature milk. Gestational age is indicated as “T” (term) or “PT” (preterm). Each number refers to a different milk sample. Standard molecular weights are indicated on the left. Each box shows the blot reactivity to a specific anti-TLR antibody and, on the lower side, the same blot incubated with an anti- β -actin antibody.

Article Snippet: Anti-TLR1 , Polyclonal rabbit antibody, directed against a recombinant peptide from human TLR1 (STJ25862) , St John's Laboratory , 1 : 500.

Techniques: Western Blot, Incubation

Journal: Journal of Immunology Research

Article Title: Analysis of Toll-Like Receptors in Human Milk: Detection of Membrane-Bound and Soluble Forms

doi: 10.1155/2019/4078671

Figure Lengend Snippet: Immunoblot of TLR1/4/6 in colostral skimmed milk. Gestational age is indicated as “T” (term) or “PT” (preterm). Each number refers to a different milk sample. Standard molecular weights are indicated on the left. Each box shows the blot reactivity to a specific anti-TLR antibody and, on the lower side, the same blot incubated with anti- β -actin Ab.

Article Snippet: Anti-TLR1 , Polyclonal rabbit antibody, directed against a recombinant peptide from human TLR1 (STJ25862) , St John's Laboratory , 1 : 500.

Techniques: Western Blot, Incubation

Journal: Journal of Immunology Research

Article Title: Analysis of Toll-Like Receptors in Human Milk: Detection of Membrane-Bound and Soluble Forms

doi: 10.1155/2019/4078671

Figure Lengend Snippet: Immunoblot of TLR1/6 in mature skimmed milk. Gestational age is indicated as “T” (term) or “PT” (preterm). Each number refers to a different milk sample. Standard molecular weights are indicated on the left. Each box shows the blot reactivity to a specific anti-TLR antibody and, on the lower side, the same blot incubated with anti- β -actin Ab.

Article Snippet: Anti-TLR1 , Polyclonal rabbit antibody, directed against a recombinant peptide from human TLR1 (STJ25862) , St John's Laboratory , 1 : 500.

Techniques: Western Blot, Incubation

Journal: ISRN Oncology

Article Title: Upregulation of TLR1, TLR2, TLR4, and IRAK-2 Expression During ML-1 Cell Differentiation to Macrophages: Role in the Potentiation of Cellular Responses to LPS and LTA

doi: 10.5402/2012/641246

Figure Lengend Snippet: Expression of TLR mRNA expression in ML-1 cells. Six million cells were incubated in the presence TPA of (5 nM) or vehicle (DMSO) for indicated times. Total RNAs were extracted from cell samples using Trizol reagents, purified with the Qiagen RNAeasy mini kit and analyzed by microarray analysis using human Genome U133 2.0 PlusArray Chip. Six replicates including three controls (DMSO-treated) and three TPA-treated ML-1 cells samples were examined. The levels of (a) TLR1, TLR2, TLR4 and TLR8 and (b) IRAK-2 mRNAs were determined by examination of the microarray analysis data.

Article Snippet: The Alexa Fluor 488-labeled mouse anti-human Abs against TLR1, TLR4, CD14, and Alexa Fluor 488-labeled isotype controls (mouse IgG1) were obtained from Molecular Probes.

Techniques: Expressing, Incubation, Purification, Microarray

Journal: ISRN Oncology

Article Title: Upregulation of TLR1, TLR2, TLR4, and IRAK-2 Expression During ML-1 Cell Differentiation to Macrophages: Role in the Potentiation of Cellular Responses to LPS and LTA

doi: 10.5402/2012/641246

Figure Lengend Snippet: TPA induces upregulation of TLR1 and TLR4 proteins expression in human myeloid ML-1 cells. Three millions cells were incubated with 5 nM TPA for various times (24 hrs, 3 days, and 6 days). (a) Cells were then harvested, washed with PBS, fixed with formaldehyde (2%), permeabilized with ice methanol, and stained using (Alexa-Fluor-488-) labeled anti-human TLR1 or TLR4 antibodies. The levels of TLR1 or TLR4 protein expression in TPA-treated ML-1 cells were determined by FACS-Scan analysis. Negative controls were stained with isotype-matched- (Alexa-Fluor 488) conjugated IgG and compensation was adjusted using the single-stained cell samples. The fluorescence intensities were determined using Cellquest software (Becton Dickinson, Bedford, MA). Data were analyzed by ModFILT statistical software. The figure depicts representative results from one of three replicate experiments. (b) After transfer onto microscope slides, cells were washed with PBS, fixed with formaldehyde (2%), permeabilizes with PBS containing 0.25% Triton X-100, and stained using (Alexa-Fluor 488 labeled anti-human TLR1 or TLR4 antibodies). The levels of TLR1 or TLR4 protein expression were examined using fluorescent microscope.

Article Snippet: The Alexa Fluor 488-labeled mouse anti-human Abs against TLR1, TLR4, CD14, and Alexa Fluor 488-labeled isotype controls (mouse IgG1) were obtained from Molecular Probes.

Techniques: Expressing, Incubation, Staining, Labeling, Fluorescence, Software, Microscopy

Journal: ISRN Oncology

Article Title: Upregulation of TLR1, TLR2, TLR4, and IRAK-2 Expression During ML-1 Cell Differentiation to Macrophages: Role in the Potentiation of Cellular Responses to LPS and LTA

doi: 10.5402/2012/641246

Figure Lengend Snippet: To determine the role of mitogen-activated protein kinase signal in TPA-induced TLR1 and TLR4 expression in ML-1 cells. Three millions cells were pretreated with 10 μ M U0126 for 10 minutes followed by incubation with 5 nM TPA for 24 hours. Cells were then harvested, washed, fixed, permeabilizes, and stained using (Alexa-Fluor-488-) labeled anti-human TLR1 or TLR4 antibodies. The levels of TLR1 or TLR4 protein expression in TPA-treated ML-1 cells were determined by FACS-Scan analysis. Negative controls were stained with isotype-matched- (Alexa-Fluor 488) conjugated IgG and compensation was adjusted using the single-stained cell samples. The fluorescence intensities were determined using Cellquest software (Becton Dickinson, Bedford, MA). Data were analyzed by ModFILT statistical software. The figure depicts representative results from one of three replicate experiments.

Article Snippet: The Alexa Fluor 488-labeled mouse anti-human Abs against TLR1, TLR4, CD14, and Alexa Fluor 488-labeled isotype controls (mouse IgG1) were obtained from Molecular Probes.

Techniques: Expressing, Incubation, Staining, Labeling, Fluorescence, Software