跟著細胞去旅行遇見最美好的 data T cell T lymphocyte T,,,T T (Cytotoxic T cell / Tc) perforin, granzymes, granulysin T (helper T cells (Th)) Th1, Th2, Th9

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1 跟著細胞去旅行遇見最美好的 data 旅行的意義 Th9 Th1 品嚐了夜的巴黎踏過下雪的北京累計了許多飛行 Th2 Th22 用心挑選紀念品蒐集了地圖上每一次的風和日麗於是遇見最美好的 data Treg Cytotoxic T Th17 Tfh Unimed

2 跟著細胞去旅行遇見最美好的 data T cell T lymphocyte T,,,T T (Cytotoxic T cell / Tc) perforin, granzymes, granulysin T (helper T cells (Th)) Th1, Th2, Th9, Th17, Th22 Tfh cells,, B, (macrophage) (eosinophil) (neutrophil) T (regulatory T cells (Tregs)) T, T CD4 + Treg, Natural Treg, ntreg, CD25 FoxP3; Induce Treg, itreg,cd25 FoxP3 Type of Cell Cytotoxic Th1 Th2 Main Function Kill virus-infected cells Activate microbicidal function of infected macrophages, and help B cells to produce antibody Help B cells and switch antibody isotype production Pathogens Targeted Viruses and some intracellular bacteria Intracellular pathogens Parasites Harmful Function Transplant rejection Autoimmune disease Allergy, asthma Extracellular Markers CD8 CD4 CXCR3 CD4 CCR4, Crth2 (human) Differentiation Cytokines IFN-γ, IL-2, IL-12, IL-18, IL-27 IL-4, IL-2, IL-33 Effector Cytokines IFN-γ, TNF, LT-α IFN-γ, LT-α, TNF IL-4, IL-5, IL-6, IL-13 Transcription Factors T-bet, Stat1, Stat6 GATA3, Stat5, Stat6 * :BD :Key marker 2

3 arriv al 研究 T 細胞的流式技術 Th9 Allergy CD4 IL-4, TGF-β IL-9, IL-10 GATA3, Smads, Stat6 2 Intracellular cytokine staining 3 BD Cytometric Bead Array (CBA) 利用血清培養液同時定性定量多種細胞激素 4 可以利用 BrdU 和 Annexin V 或其他試劑來偵測細胞表面抗原鑑定偵測單一細胞內細胞激素的表現 Cell proliferation and Apoptosis 5 轉錄因子和磷酸化蛋白偵測偵測單一細胞內磷酸化蛋白的表現 Tfh Treg Regulate development Enhance neutrophil response production by B cells Parasites Surface marker staining Th17 T cell proliferation and enhanced IgG and IgE 1 of antigen speciﬁ c B cell development and antibody Immune regulation production Fungi and extracellular bacteria Organ-speciﬁc autoimmune disease CD4, CCR6 TGF-β, IL-6, IL-1, IL-21, IL23 IL-17A, IL-17F, IL-21, IL-22, IL-26, TNF, CCL20 RORγt, RORα, Stat3 Autoimmune disease Autoimmune disease, cancer CD4, CXCR5 CD4, CD25 IL-12, IL-6 TGF-β, IL-12 IL-21 TGF-β, IL-10 Bcl-6, MAF FoxP3, Smad3, Stat5 3

4 跟著細胞去旅行遇見最美好的 data (surface marker staining), Phenotyping of Cells with Unique Surface Profiles The 13-color panel below examines memory and activation status of multiple T-cell subsets Lysed whole blood (A) T-cell subsets (B) Tregs and memory cells. 4

5 arrival (Techniques for the Detection of Secreted Cytokines), T,, Intracellular Cytokine Staining (ICS), BD Cytometric Bead Array (CBA) (capture antibody),, PE (detection antibody), PE, 1,, ELISA 1 1, ICS/CBA/ELISA CAPABILITY CBA ELISA ICS O X O X X O O O X 5

6 跟著細胞去旅行遇見最美好的 data (Techniques for the Detection of Secreted Cytokines) 1 CBA 2 Intracellular Cytokine Staining, Th1/Th2/Th17 + IL-2, IL-12, neutralizing mab to IL-4 TH1 Polarization T cells co-stimulated with CD3/CD28 + IL-2, IL-4, neutralizing mab to IFN-γ + IL-2, IL-6, IL-1β, TGF-β, IL-23, and a neutralizing mab to IL-4 and IFN-γ TH2 Polarization TH17 Polarization 1 CBA beads sample detector antibody 2-3,! CBA kit, Th1, Th2, Th17 polarized,, ICS Cat. No. Description contents size Human Human Th1/Th2 Cytokine Kit IL-2, IL-4, IL-5, IL-10, TNF, IFN-γ 80 tests Human Th1/Th2 Cytokine Kit II IL-2, IL-4, IL-6, IL-10, TNF, IFN-γ 80 tests Human Th1/Th2/Th17 Kit IL-2, IL-4, IL-6, IL-10, TNF, IFN-γ, IL-17A 80 tests Mouse Mouse Th1/Th2 Cytokine Kit IL-2, IL-4, IL-5, IFN-γ, TNF 80 tests Mouse Th1/Th2/Th17 Kit IL-2, IL-4, IL-6, IFN-γ, TNF, IL-17A, IL tests Non-human Primate (Baboon, Cynomolgus, Rhesus) Non-human Primate Th1/Th2 Cytokine Kit IL-2, IL-4, IL-5, IL-6, TNF, IFN-γ 80 tests * flex set,, 30 6

arrival 2 Intracellular Cytokine Staining

Naive/Memory T Cell Panel 50 Tests 560759

560767 Mouse Th17/Treg Phenotyping Kit 50

7 arrival 2 Intracellular Cytokine Staining (ICS) Stimulate fixed and permeabilized Stain + monensin 6h FACS Analysis Th17 polarized BD GolgiStop (monensin) inhibitor, BD Cytofix/Perm (ant-cd4, IL-17A, IFN-γ, IL-4) BD phenotyping kit Cat. No. Description species size Human Th1/Th2/Th17 Phenotyping Kit 50 Tests HumanTh1/Th17 Phenotyping Kit 50 Tests Hu HumanTh17/Treg Phenotyping Kit 50 Tests Human Naive/Memory T Cell Panel 50 Tests Mouse Th1/Th17 Phenotyping Kit 50 Tests Mouse Th17/Treg Phenotyping Kit 50 Tests Ms Mouse Th1/Th2/Th17 Phenotyping Kit 50 Tests Mouse Naive/Memory T Cell Panel 50 Tests 7

8 跟著細胞去旅行遇見最美好的 data Intracellular Signaling Tools for Analysis of T-cell signaling,, Adaptive immune system T,,Naïve CD4 T Th1, Th2, Th9, Th17, Tfh Tregs,, T BD transcription factor buffer set (Cat. No /562725), ( CD4), BD Phosflow,, BD Phosflow Human T Cell Activation Kit (cat.#560750) CD4 CD8 T, Cat. No. Description Content size BD Phosflow BD Phosflow Alexa Fluor 647 Conjugates human antibodies: p38 MAPK (pt180/y182), ERK (pt202/204), Stat1 (py701), Stat3 (py705), Stat5 (py694), Stat6 (py641) Phosflow Perm Buffer III Stain Buffer (FBS) human control cells ( Stimulated/ unstimulated human T cells ) Human T Cell BD Phosflow Human T Cell antibody Cocktail Activation Kit CD3/CD4/CD8 Compensation Reagents Phosflow Lyse/Fix Buffer (5X) 50 Tests, T cell All in one : cocktail ( CD3/CD4/CD8 ) cocktail, kit buffer protocol, Kit positive / negative control cells 8

arrival T Regulatory T cells (Tregs) T (Tregs) T, Tregs, (multiple sclerosis) (active

inducible Tregs (itregs) ntregs, CD4CD25 Foxp3 Foxp3 Tregs marker, Tregs, Tregs, CD127(IL-7Rα

Th3 CD4+CD25+ from Phenotype CD4+CD25 int/high, CD127 low CD4 + CD25 - CD25- precursors Other

-variable, CD45RB low, FoxP3 + Suppression Contact dependent, granzyme Through cytokines

and T effector cells T effector cells Not yet Identified CD28 Involvement Thymic development

Suppression of autoreactive T Mucosal immunity, Mucosal immunity, cells inflammatory response

9 arrival T Regulatory T cells (Tregs) T (Tregs) T, Tregs, (multiple sclerosis) (active rheumatoid arthritis) (type 1 diabetes) Tregs,Tregs, CD4+Tregs natural Tregs (ntregs) inducible Tregs (itregs) ntregs, CD4CD25 Foxp3 Foxp3 Tregs marker, Tregs, Tregs, CD127(IL-7Rα ) CD4 CD25, CD127 Tregs Treg subsets Natural Treg (ntregs) Induced Tregs (itregs) ntreg Tr1 Th3 CD4+CD25+ from Phenotype CD4+CD25 int/high, CD127 low CD4 + CD25 - CD25- precursors Other associated markers CTLA4 +, GITR +, FoxP3 +, CD127 low CD25, CD45RB low, FoxP3 - CD25 -variable, CD45RB low, FoxP3 + Suppression Contact dependent, granzyme Through cytokines produces Through cytokines, B-dependent, makes TGF-β IL-10 produces TGF-β Target cells APC and T effector cells T effector cells Not yet Identified CD28 Involvement Thymic development and Not for development or maintenance in periphery function Not involved In vivo Role Suppression of autoreactive T Mucosal immunity, Mucosal immunity, cells inflammatory response inflammatory response In vitro Expansion Expandable using TCR/CD28 stimulation and IL-2 CD3, IL-10, rehnoic acid CD3, TGF-β 9

跟著細胞去旅行遇見最美好的 data T Regulatory T cells (Tregs) Detection of human Tregs by

Human PBMCs were stained for surface markers using the following fluorescent

After washing, the cells were fixed and permeabilized using the BD Pharmingen

Gated Tregs (colored blue) have a CD4+ CD25high CD127dim FoxP3 + phenotype.

#648956) CD3/CD28 stimulated PBMCs Stimulated PBMCs in the presence of Tregs The BD

10 跟著細胞去旅行遇見最美好的 data T Regulatory T cells (Tregs) Detection of human Tregs by intracellular flow cytometry. Human PBMCs were stained for surface markers using the following fluorescent antibodies: CD4 FITC, CD25 BV421, and CD127 Alexa Fluor 647. After washing, the cells were fixed and permeabilized using the BD Pharmingen transcription factor buffer set (Cat.#562574). Cells were then stained with FoxP3. Gated Tregs (colored blue) have a CD4+ CD25high CD127dim FoxP3 + phenotype. effector T cell Tregs BD Regulatory T-Cell Function Kit (Cat.#648956) CD3/CD28 stimulated PBMCs Stimulated PBMCs in the presence of Tregs The BD FastImmune kit (Cat.# ) measures expression of the activation markers CD154 and/or CD69 on effector T cells. Reduced expression of these markers in the presence of Tregs can indicate Treg suppressive capacity. 10

11 arrival Cat. No. Description content size BD Human Regulatory T Cell Sorting Kit Treg sorting cocktail, CD45RA FITC; CD127 Alexa Fluor 647; CD25 PE; CD4 PerCP- Cy Tests (1 Test = 1 x 10 8 cells) BD Human Regulatory T Cell Cocktail FITC CD4 FITC; Cd25 PE-Cy7; CD127Alexa Fluor Tests BD Anti-Human FoxP3 Staining Kit - Alexa Fluor 647, FoxP3, CD4, CD25 Foxp3 Alexa Fluor 647; CD25 PE; CD4 FITC; Foxp3 buffer set 100 Tests BD Anti-Human FoxP Staining Kit - PE, FoxP3, CD4, CD25 Foxp3 PE; CD25 APC; CD4 FITC 100 Tests BD Regulatory T-Cell Function Kit Regulatory T-Cell function cocktail (CD3 PerCP-Cy 5.5, CD4 FITC, CD25 PE); CD69 PE-Cy 7; CD154 APC; EDTA. 50 Tests BD Human FoxP3 Buffer Set Two part buffer system for detection of human FoxP3 100 Tests BD Mouse FoxP3 Buffer Set Mouse Foxp3 fixation and permeabilization buffers 100 Tests BD Transcription Factor Buffer Set Fixation/permeabilization buffer, diluent buffer, perm/wash buffer 100 Tests 11

12 跟著細胞去旅行遇見最美好的 data T (T cell receptor, TCR) T T (T cell receptor, TCR), (antigen-presenting cells;apc) MHC, T cells, T TCR α chain β chain, T γ chain δ chain, T γδ T αβ T, γδ T,, αβ T cell, γδ T MHC, γδ T, T T TCR, γδ T Vγ9+ /Vδ2+, γδ T, TCR TCR Vγ9 PE (Cat# BD ) CD3 FITC, BD TCR marker Hunam Mouse Rat TCR β TCR Vα11.1, 11.2[b,d] TCR Vβ9 TCR αβ TCR Vβ3 TCR Vα2 TCR Vβ10[b] TCR γδ TCR Vβ8 TCR Vα3.2[b,c] TCR Vβ11 TCR Vγ9 TCR Vα8 TCR Vβ12 TCR Vδ2 TCR Vα8.3 TCR Vβ13 TCR Cb1 TCR Vβ2 TCR Vβ14 TCR αβ TCR Vβ3 TCR Vβ17[a] TCR γδ TCR Vβ4 TCR Vγ2 TCR Vβ5.1, 5.2 TCR Vγ3 TCR Vβ6 TCR Vδ 6.3/2 TCR Vβ7 TCR Vδ4 TCR Vβ8 TCR β Chain TCR Vβ8.1, 8.2 TCR γδ TCR Vβ8.3 TCR Vγ1.1 TCR DO Clonotypic 12

arrival Next Generation Sequencing (NGS) TCR, TCR, T cells,, T SMARTer Human sctcr a/b Profiling Kit 5'-RACE, FACS single T cells TCR libraries, low-abundance TCR variants a/b pairing,pcr indexes,

TCR-beta sequencing ready library 10 ng 3 µg RNA (obtained from peripheral blood) 50 10,000 purified T cells T-Cell Receptor (TCR) alpha chain beta chain T-Cell TCR repertoire, T Cell T-Cell α-β

13 arrival Next Generation Sequencing (NGS) TCR, TCR, T cells,, T SMARTer Human sctcr a/b Profiling Kit 5'-RACE, FACS single T cells TCR libraries, low-abundance TCR variants a/b pairing,pcr indexes, multiplex 96 SMARTer Human TCR a/b Profiling Kit T 5'-RACE SMART-Seq v4 LNA T Cell Receptor (TCR) mrna transcript variable regions, TCR multiplex PCR TCR profiling, bias amplification TCR-alpha TCR-beta sequencing ready library 10 ng 3 µg RNA (obtained from peripheral blood) 50 10,000 purified T cells T-Cell Receptor (TCR) alpha chain beta chain T-Cell TCR repertoire, T Cell T-Cell α-β chain Brand Cat. No. Description size Rxns SMARTer Human sctcr a/b Profiling Kit Rxns Clontech Rxns SMARTer Human TCR a/b Profiling Kit 48 Rxns Rxns 13

跟著細胞去旅行遇見最美好的 data T cell culture (in vitro assay), (proliferation, adhesion, or migration ) NA/LE antibody BD (no azide, low endotoxin, NA/LE), T,azide (in vitro/in vivo assay) (Endotoxin level is

14 跟著細胞去旅行遇見最美好的 data T cell culture (in vitro assay), (proliferation, adhesion, or migration ) NA/LE antibody BD (no azide, low endotoxin, NA/LE), T,azide (in vitro/in vivo assay) (Endotoxin level is 0.01 EU/µg ( ng/µg) Cat. No. Description clone size Purified NA/LE Mouse Anti-Human CD3 UCHT1 0.5 mg Purified NA/LE Mouse Anti-Human CD28 CD mg Purified NA/LE Hamster Anti-Mouse CD3e 145-2C mg Purified NA/LE Hamster Anti-Mouse CD mg Purified NA/LE Mouse Anti-Rat CD3 G mg Purified NA/LE Mouse Anti-Rat CD28 JJ mg Anti-CD3 mab GMP grade GMP, ex vivo cell culture OKT3 Clone, mouse IgG2a isotype GMP Grade Grade Cat. No. Description size T210 Anti-CD3 mab GMP grade 1 mg/1 ml 14

arrival RetroNectin GMP Grade GMP, ex vivo

15 arrival RetroNectin GMP Grade GMP, ex vivo cell culture Anti-CD3/IL-2 T cells, T cell population naïve T cells CH-296 with Carbonate US FDA (Drug Master File) Retronectin (RN), 2! Cat. No. Description size T201 Tarkara RetroNectin GMP grade 2.5 mg CultiLife Cell Culture Bags " ", flask CultiLife 215 CultiLife Eva CultiLife Medium,CultiLife Flask Cat. No. Description size (cm 2 ) (ml) FU0005 CultiLife 215 Culture Bag* ~300 FU0010 GT-T610 (CultiLife Eva) Culture Bag ~1000 * coating anti-cd3 & Retronectin 15

tw 02-2720-2215 03-6684-586 04-2463-3591

16 T-Cell Research Brochure CD4 + T Cell Differentiation poster Unimed U070629

,, flow, cell viability dye cell viability dye PI, 7-AAD, Fixable Viability Stain (FVS) Reagents,FVS reagents Amine,,FVS ;, FVS, FVS 10-20, A. BD Cyto

DM 8 7 (Cat.562253) 7 1000 2017/6/30 TEL (02)2720-2215 TEL (05)2844-162 TEL (03)6684-586 TEL (06)2890-665 TEL (04)2463-3591 TEL (07)3470-143 免付費客服專線 01 ,, flow, cell viability dye cell viability dye PI,

跟著細胞去旅行 遇見最美好的 data T cell T lymphocyte T,,,T T (Cytotoxic T cell / Tc) perforin, granzymes, granulysin T (helper T cells (Th)) Th1, Th2, Th9

跟著細胞去旅行遇見最美好的 data T cell T lymphocyte T,,,T T (Cytotoxic T cell / Tc) perforin, granzymes, granulysin T (helper T cells (Th)) Th1, Th2, Th9