anti il17a pe  (Elabscience Biotechnology)

Journal: bioRxiv

Article Title: Allosteric inhibition of CXCR1 and CXCR2 abrogates Th2/Th17-associated Allergic Lung Inflammation in Mice

doi: 10.1101/2024.05.13.593638

Figure Lengend Snippet: ( A ) Rorc mRNA expressions in the BALF cells after CDE challenge in CDE-SCM and CDE-MCM mice at 28 h post-CDE challenge. ( B ) Il17 and Rorc mRNA expressions in the BALF cells in CDE-MCM at 28 h post-CDE challenge. ( C ) Il17 mRNA expressions in the lung in CDE-MCM at 28 h post-CDE challenge. ( D-E ) The number of RorγT T-cells ( D ) and IL17 secreting Th17-cells ( E ) in the lung in CDE-MCM at 72 h post-CDE challenge. BALF; bronchoalveolar lavage fluid CDE; cat dander extract CDE-MCM; CDE-multiple challenge model RorγT; retinoic acid–related orphan receptor-gamma t Th17; T helper 17

Article Snippet: Lung single cells were pre-incubated with TruStain FcX solution (BioLegend, San Diego, CA) for 10 mins at 4°C, and stained with fluorophore-conjugated antibodies Fixable Viability Dye eFluor 780 (eBioscience, San Diego, CA), CD45 monoclonal antibody-pacific orange (Thermo Fisher Scientific, MA, USA, #MCD4530, clone 30-F11), anti-mouse CD3-Alexa Fluor 700 (Biolegend, #100216, clone 17A2), rat anti-mouse CD4-Brilliant Violet 520 (BD Biosciences, #563106, clone RM4.5), PE rat anti-mouse CD181 (CXCR1) (BD Biosciences, San Jose, CA, #566383, clone U45-632), BV711 Rat Anti-Mouse CD182 (CXCR2), (BD Biosciences, #747812, clone V48-2310), BV786 rat anti-mouse Siglec-F (BD Biosciences, #740956, clone E50-2440), Spark YG 593 anti- mouse Ly-6G Antibody (Biolegend, #127668, clone 1A8), Spark NIR™ 685 anti-mouse/human CD11b Antibody (Biolegend, #101278, clone M1/17) at 1:100 dilution in flow stain buffer containing FBS for 30 mins at 4°C, then cells were permeabilized with Fixation/Permeabilization kit (BD Biosciences) according to the protocol from vendor and stained with intracellular cytokine specific antibodies Rat anti-mouse Rorγt- Brilliant Violet-650 (BD Biosciences, #564722, clone Q31-378), mouse anti-GATA3 Alexa Fluor F488 (BD Bioscience #560163, clone L50-823,) Rat anti-mouse IL4-PE/Cyanine7 (BD Biosciences, #560699, clone 11B11), anti-mouse/anti-human IL5-Allophycocyanin (APC) (BD Biosciences, #554396, clone TRFK5), IL13 monoclonal antibody ((eBio13A), Brilliant Ultra Violet 805, eBioscience), Rat Anti-Mouse IL17A-PE (BD Biosciences, # 559502, clone TC11-18H10), and rat anti-mouse IL23 p19 Alexa Fluor 647 (BD Biosciences, # 565317, clone N71-1183) for 45 mins at 4°C.

Techniques:

Journal: bioRxiv

Article Title: Inflammation induced T h 17 cells synergize with the inflammation-trained microbiota to mediate host-resiliency against intestinal injury

doi: 10.1101/2024.03.25.586435

Figure Lengend Snippet: A) IL-17 FM mice, which permanently activate GFP upon IL-17 expression, were subjected to GI or iv Cr or PBS. They were allowed to recover and sacrificed at week 6. Live lamina propria mononuclear cells (LPMCs) were gated from mice in (A) into groups as indicated and their (B) relative fractions and (C) absolute numbers were quantified. D) Intestinal CD4 + T-cells from mice in (A) were gated as shown, with further gating on GFP + (T h 17) (middle and right columns). E) T h 17 T RM defined as CD4 + GFP + CD44 + CD69 + from (D) and F) IFNɣ + T h 17 T RM were enumerated. G) The fractions of T h 17 T-effector memory cells (CD4 + GFP + CD44 + CD69 - ) in spleen of mice in (A) were quantified. H) Wild-type mice were subject to PBS or 2.5% DSS for 1 week and then allowed to recover. LPMCs were assessed by flow cytometry at week 6 and the fraction (left panel) and absolute numbers (right panel) of T h 17 T RM was quantified * to **** p < 0.01-0.00001 by ANOVA (B-G) or students t-test (H). Data are representative of 2-6 experimental replicates, with 3-9 mice per group, per experiment.

Article Snippet: The following antibodies were used for flow cytometry: CD3ε BV 510 (BioLegend, clone 145-2C11), CD44 BV 421 (BioLegend, clone IM7), CD8β PerCP-eFluor 710 (eBiosciences, clone H35-17.2), CD8β PerCP Cy5.5 (BioLegend, clone YTS156.7.7), IFNγ APC (BioLegend, clone XMG1.2), IFNγ PE (BioLegend, clone XMG1.2), CD4 FITC (Thermo Fisher, Waltham, MA, Clone GK1.5), CD4 PE Dazzle (BioLegend, clone GK1.5), CD4 BV 650 (BioLegend, clone RM4-5), CD69 PE-Cy7 (BioLegend, clone H1.2F3), IL-17A PE, IL-17A APC, IL-17 FITC (BioLegend, clone TC11-18H10.1), CD45 BV 605 (BioLegend, clone 30-F11), Annexin V CF405M (Biorbyt), Fix Viability eFluor 780 (Thermo Fisher)

Techniques: Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: Inflammation induced T h 17 cells synergize with the inflammation-trained microbiota to mediate host-resiliency against intestinal injury

doi: 10.1101/2024.03.25.586435

Figure Lengend Snippet: A) J H -/- mice were given convalescent (con) serum from wild type mice that had cleared GI Cr , and subject to GI Cr or PBS and (B) weight change, (C) disease activity index (DAI) and (D) survival were measured. E) RAG1 -/- mice were reconstituted with wild-type total CD4 + T-cells and then subject to GI Cr or PBS. Because IgG is required to clear Cr , all mice were also given convalescent (con) serum from mice that had previously cleared GI Cr . All mice were then subject to 2.5% DSS at week 6 and F) weight change and G) survival were measured. H) Wild-type mice were subject to PBS or GI Cr , allowed to clear and then subject to FTY 720 at week 6 followed by 2.5% DSS at week 8. Control mice received PBS without FTY 720. I) Weight change and (J) survival were measured. K) IL-17 FM mice were subject to PBS or GI Cr +/- FTY 720 as in (H) and T h 17 T RM in the colon were enumerated at week 8 (prior to DSS). * to *** p < 0.01-0.0001 by ANOVA (B, C, F, I, K) or Kaplan-Meier (D, G, J). Data are representative of 2-6 experimental replicates, with 3-5 mice per group, per experiment.

Article Snippet: The following antibodies were used for flow cytometry: CD3ε BV 510 (BioLegend, clone 145-2C11), CD44 BV 421 (BioLegend, clone IM7), CD8β PerCP-eFluor 710 (eBiosciences, clone H35-17.2), CD8β PerCP Cy5.5 (BioLegend, clone YTS156.7.7), IFNγ APC (BioLegend, clone XMG1.2), IFNγ PE (BioLegend, clone XMG1.2), CD4 FITC (Thermo Fisher, Waltham, MA, Clone GK1.5), CD4 PE Dazzle (BioLegend, clone GK1.5), CD4 BV 650 (BioLegend, clone RM4-5), CD69 PE-Cy7 (BioLegend, clone H1.2F3), IL-17A PE, IL-17A APC, IL-17 FITC (BioLegend, clone TC11-18H10.1), CD45 BV 605 (BioLegend, clone 30-F11), Annexin V CF405M (Biorbyt), Fix Viability eFluor 780 (Thermo Fisher)

Techniques: Activity Assay, Control

Journal: bioRxiv

Article Title: Inflammation induced T h 17 cells synergize with the inflammation-trained microbiota to mediate host-resiliency against intestinal injury

doi: 10.1101/2024.03.25.586435

Figure Lengend Snippet: A) The microbiota of IL-17 FM mice was assessed at baseline and 2 weeks after clearing GI Cr . B) Schematic of fecal microbiota transfer (FMT) experiments with 4 groups of mice ( Cr (GI), Cr (GI) + Abx, Abx + FMT ( Cr ), and Ctrl + Abx). C) Weight change (left panel), colon length (middle panel) and survival (right panel) in groups from (B) with 2.5% DSS. D) Schematic of co-housing experiments showing separately housed (SH) and cohoused (CH) cohorts of controls (Ctrl) and Cr (GI) mice. After CH, mice in this cohort were separated into cages based on prior history of Cr or PBS. All cohorts were then subject to 2.5% DSS and (E) their survival was determined. F, G) Lamina propria mononuclear cells were assessed by flow cytometry after 2 weeks of antibiotics in the indicated groups from (B), and (F) the ratio of the fraction of CD4 + T EM /T RM was quantified and (G) enumerated. H) Mice were left unmanipulated (Control) or subject to GI Cr , allowed to recover, and then monitored (GI Cr ) or treated with 2 weeks of antibiotics (GI Cr + Abx). At week 8 post-infection, lamina propria mononuclear cells were assessed for expression of Annexin V by flow cytometry (left panel) and the data were quantified (right panel). The gating scheme is Live → CD3 + CD4 + → GFP + → CD44 + CD69 + → Annexin V. * to *** p < 0.01-0.0001 by Kaplan-Meier (C, E), students t-test (F) or ANOVA (C, G). Data are representative of 3 (B, C), 2 (D-F), or one (G, H) experimental replicates with 3 to 7 mice per group.

Article Snippet: The following antibodies were used for flow cytometry: CD3ε BV 510 (BioLegend, clone 145-2C11), CD44 BV 421 (BioLegend, clone IM7), CD8β PerCP-eFluor 710 (eBiosciences, clone H35-17.2), CD8β PerCP Cy5.5 (BioLegend, clone YTS156.7.7), IFNγ APC (BioLegend, clone XMG1.2), IFNγ PE (BioLegend, clone XMG1.2), CD4 FITC (Thermo Fisher, Waltham, MA, Clone GK1.5), CD4 PE Dazzle (BioLegend, clone GK1.5), CD4 BV 650 (BioLegend, clone RM4-5), CD69 PE-Cy7 (BioLegend, clone H1.2F3), IL-17A PE, IL-17A APC, IL-17 FITC (BioLegend, clone TC11-18H10.1), CD45 BV 605 (BioLegend, clone 30-F11), Annexin V CF405M (Biorbyt), Fix Viability eFluor 780 (Thermo Fisher)

Techniques: Flow Cytometry, Control, Infection, Expressing

Journal: iScience

Article Title: Targeting abatacept-resistant T-helper-17 cells by aldehyde dehydrogenase inhibition

doi: 10.1016/j.isci.2023.108646

Figure Lengend Snippet:

Article Snippet: PE anti-mouse IL17A , Thermo Fisher , Cat#12-7177-81 ;RRID: AB_763582.

Techniques: Functional Assay, Recombinant, Cell Counting, Reverse Transcription, Sequencing, Knock-Out, Software