mouse th17 cell differentiation kit  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: Peripheral blood mononuclear cell low molecular mass protein 7 in acute ischemic stroke: vertical change from admission to discharge and correlation with disability, stroke recurrence, and death

doi: 10.3389/fimmu.2024.1296835

Figure Lengend Snippet: Characteristics at admission.

Article Snippet: Then those proportions were determined by flow cytometric (FCM) analysis (FlowX Human Th1, Th2, and Th17 Cell Multi-Color Flow Cytometry Kits, No. Cat. FMC009B, FMC011B, and FMC007B, Bio-Techne, China).

Techniques:

Journal: Frontiers in Immunology

Article Title: Peripheral blood mononuclear cell low molecular mass protein 7 in acute ischemic stroke: vertical change from admission to discharge and correlation with disability, stroke recurrence, and death

doi: 10.3389/fimmu.2024.1296835

Figure Lengend Snippet: Elevated PBMC LMP7 at admission was correlated with decreased Th2 cells and increased Th17 cells, CRP, and NIHSS score in AIS patients. The correlation of PBMC LMP7 at admission with Th1 cells (A) , Th2 cells (B) , Th17 cells (C) , CRP (D) , CRP ≥5 mg/L (E) , NIHSS score (F) , and stroke severity classified by NIHSS score (G) .

Article Snippet: Then those proportions were determined by flow cytometric (FCM) analysis (FlowX Human Th1, Th2, and Th17 Cell Multi-Color Flow Cytometry Kits, No. Cat. FMC009B, FMC011B, and FMC007B, Bio-Techne, China).

Techniques:

Journal: JCI Insight

Article Title: Acquired immunostimulatory phenotype of migratory CD103 + DCs promotes alloimmunity following corneal transplantation

doi: 10.1172/jci.insight.182469

Figure Lengend Snippet: ( A ) Experimental schematic. Allogeneic (C57BL/6J) corneal grafts were transplanted into BALB/C LR and HR RAG –/– recipient mice. Two weeks after transplantation, CD103 + DC1 were sorted from the migratory compartment, as described above. CD103 + DC1 from LR recipients were incubated with IFN-γ, and those from HR RAG –/– were left untreated. ( B – E , F , and G ) After 12 hours, cells were analyzed by flow cytometry for the expression of regulatory markers ( B – E ) and supernatant for the secretion of IL-10 ( F ) and IL-12 ( G ). Each symbol in B – E indicates an individual mouse. ( H ) iDC1 were sorted by FACS and preincubated with or without IFN-γ and then cocultured with in vitro–differentiated Th1 CD4 + T cells with or without anti–PD-L1 blocking antibodies. ( I and J ) The intracellular expression of IFN-γ by CD4 + T cells was analyzed by flow cytometry, and the mean (± SEM) of frequencies ( I ) and expression (MFI) ( J ) were measured. Results show 2 sets of triplicates. ( K ) iDC1 ( , A and B) were sorted by FACS and preincubated with or without IFN-γ. iDC1 were transwell plated (upper) with in vitro–differentiated Th1 CD4 + T cells (lower). Additionally, mouse recombinant IL-12 or anti-IL-10 were added. ( L and M ) The intracellular expression of IFN-γ by CD4 + T cells was analyzed by flow cytometry, and the mean (± SEM) of frequency ( L ) and expression (MFI) ( M ) was measured. Results show 2 sets of triplicates. ** P < 0.01, *** P < 0.001, **** P < 0.0001 (1-way ANOVA with Bonferroni correction). All results are of one experiment with no repetitions. Experimental schematics created with Biorender.com.

Article Snippet: CD4 + CD44 – CD25 – T naive T cells were double stimulated using a CellXVivo Mouse Th1 Cell Differentiation Kit (R&D Systems).

Techniques: Transplantation Assay, Incubation, Flow Cytometry, Expressing, In Vitro, Blocking Assay, Recombinant

Journal: JCI Insight

Article Title: Acquired immunostimulatory phenotype of migratory CD103 + DCs promotes alloimmunity following corneal transplantation

doi: 10.1172/jci.insight.182469

Figure Lengend Snippet: ( A ) BALB/C HR recipients were transplanted as above. Immediately after transplantation, allograft recipient mice received 2 × 10 4 of CMTMR-labeled iDC1 or PBS via SC injection. ( B ) Two weeks after transplantation, CMTMR – migratory CD103 + DC1 were sorted by FACS from the DLN and analyzed by qPCR assay, showing the expression average of most differentially functional regulatory markers. ( C – H ) The MFI of regulatory markers was measured by flow cytometry ( C – F ) or ELISA ( G and H ) and compared with DC2 from HR recipients (previous gating from above; ). ( I and J ) Intracellular expression of IFN-γ by Th1 and FoxP3 + CD25 + gated on CD3 + CD4 + T cells from the DLN of graft recipient mice. ( K and L ) Ex vivo Treg suppressive capacity on proliferating CD4 + T cells was measured. Corneal grafts were monitored for up to 8 weeks after transplant ( n = 8 group), and Kaplan-Meier curves were plotted to evaluate graft survival. Representative slit lamp photography from either LR or LR are presented. BALB/C LR recipients were transplanted as above. Immediately after transplantation, allograft recipient mice received 2 × 10 4 CMTMR-labeled iDC1 pretreated with IFN-γ or PBS (vehicle internal control) via SC injection. Two weeks after transplantation, mice were euthanized ( n = 6 group). ( M and N ) CMTMR – migratory CD103 + DC1 were sorted by FACS and analyzed by qPCR assay as above. ( O and P ) Intracellular expression of IFN-γ by Th1 ( O ) and FoxP3 + CD25 + gated on CD3 + CD4 + T cells ( P ) from the DLN of graft recipient mice was measured. ( Q ) Ex vivo Treg suppressive capacity on proliferating CD4 + T cells was measured. Additional mice were followed up ( n = 8/group) for 8 weeks with slit lamp examination to evaluate graft opacity, and Kaplan-Meier curves were plotted to evaluate graft survival. Plots represent the mean (± SEM). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (2-tailed t test). All results are of 1 experimental set of animals with no repetitions.

Article Snippet: CD4 + CD44 – CD25 – T naive T cells were double stimulated using a CellXVivo Mouse Th1 Cell Differentiation Kit (R&D Systems).

Techniques: Transplantation Assay, Labeling, Injection, Expressing, Functional Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Ex Vivo, Control