MAIT cell function is modulated by PD-1 signaling in patients with active

Size: px
Start display at page:

Download "MAIT cell function is modulated by PD-1 signaling in patients with active"

Transcription

1 MAIT cell function is modulated by PD-1 signaling in patients with active tuberculosis Jing Jiang, M.D., Xinjing Wang, M.D., Hongjuan An, M.Sc., Bingfen Yang, Ph.D., Zhihong Cao, M.Sc., Yanhua Liu, Ph.D., Jinwen Su, M.D., Fei Zhai, B.S., Ruo Wang, M.Sc., Guangyu Zhang, M.D., Xiaoxing Cheng #, M.B., Ph.D. Online Data Supplement

2 Supplementary methods (1) Human subjects One hundred and twenty-three patients with active TB, including 93 patients with pulmonary TB, 26 with tuberculous pleurisy, and 4 with tuberculous peritonitis, were recruited (Table 1), and they were diagnosed according to the 1990 edition of Diagnostic Standards and Classification of Tuberculosis, published by the American Lung Association. All patients were HIV-negative. In the 93 patients with pulmonary TB, forty-nine were sputum smear/culture-positive and 44 were sputum smear/culture-negative (Table 1). For patients with negative sputum smear/culture results, diagnosis was established by clinical symptoms, chest computed tomography and pathological examination from biopsy samples obtained through bronchoscopy and was confirmed by symptomatic and radiographic improvement after anti-tb chemotherapy. In the cohort of 49 sputum-positive patients with pulmonary TB, 30 were defined as new cases and 19 as cases of treatment failure (Table 1). TB patients who were not treated for TB or had been treated for less than two weeks at the time of blood collection were considered new cases. Nineteen TB patients who were previously treated with a standard anti-tb chemotherapy regimen without success were defined as cases of treatment failure. Patients with tuberculous pleurisy and tuberculous peritonitis were diagnosed based on clinical manifestations, computed tomography, ultrasonography, and clinical E2

3 laboratory tests, including cellular and biochemistry examinations, bacterial culture, real-time PCR specific for M. tuberculosis complex, pleural effusion or ascitic fluid ELISPOT assay using a T-SPOT.TB kit (Oxford Immunotec, Oxfordshire, United Kingdom), adenosine deaminase (ADA), pathological examination from biopsy samples, and responses to anti-tb chemotherapy. Seventy-nine healthy controls were randomly recruited from individuals undergoing annual health check-ups at the clinics of 309 Hospital using following inclusion criteria: (1) no fever, cough, or other signs of active TB; (2) normal physical examination results and normal radiography; and (3) no HIV infection. (2) Antibodies and reagents Antibodies of anti-human TCR Vα7.2-FITC or -PE (clone 3C10), anti-cd161-apc or -PE-Cy7 (clone HP-3G10), anti-cd197 (CCR7)-PE-Cy7 (clone G043H7), anti-cd279 (PD-1)-FITC (clone EH12.2H7), anti-cd160-alexa Fluor 647 (clone BY55), anti-tim-3-pe (clone F38-2E2), LEAF purified anti-human CD279 (PD-1) (clone EH12.2H7), LEAF purified anti-human/mouse/rat MR1 (clone 26.5), and LEAF purified anti-human CD28 (clone CD28.2) were purchased from BioLegend (San Diego, CA, USA). Antibodies of anti-human CD3-PE-CF594 or PE-Cy7 (clone UCHT1), anti-cd45ra-pe-cy5 (clone HI100), anti-cd45ro-fitc (clone UCHL1), anti-cd69-fitc or -PE (clone FN50), anti-cd279 (PD-1)-FITC (clone MIH4), anti-tnf-α-fitc or -PE (clone MAb11), anti-cd244-pe (clone 2-69), anti-btla-pe (clone J ), anti-tcr γδ-fitc (clone B1), the E3

4 anti-ki-67-fitc-set (clone B56), the annexin V-FITC apoptosis detection kit I, and the cytofix/cytoperm fixation/permeabilization kit were obtained from BD Biosciences (San Diego, California, USA). Antibodies of anti-ifn-γ-fitc (clone 25723) and goat anti-lag-3-pe were obtained from R&D Systems (Minneapolis, MN, USA). Antibodies of anti-cd274-apc (PD-L1) (clone MIH1) and anti-cd273-apc (PD-L2) (clone MIH18) were from ebioscience (San Diego, CA, USA). Isotype-matched control antibodies were purchased from the corresponding suppliers mentioned above to determine the background level of staining. (3) Intracellular cytokine staining For intracellular cytokine staining, cells were collected, washed, and stained first with surface markers. After being permeabilized with cytofix/cytoperm fixation/permeabilization buffer (BD Biosciences), the cells were incubated with fluorochrome-labeled anti-ifn-γ or anti-tnf-α antibody for 30 minutes at 4 C. Appropriate isotype-matched control antibodies were used to determine background levels of staining. The expression of protein was analyzed by an FC-500 flow cytometer (Beckman Coulter, Brea, CA, USA). (4) Apoptosis assay PBMCs from both patients with active TB and healthy controls were incubated with CFU/ml live BCG overnight. Cells were stained with FITC annexin V apoptosis detection kit I (BD Biosciences) by following the manufacturer s E4

5 instructions, and cell apoptosis was analyzed by flow cytometry. E5

Supplementary Table e-1. Flow cytometry reagents and staining combinations

Supplementary Table e-1. Flow cytometry reagents and staining combinations Supplementary data Supplementary Table e-1. Flow cytometry reagents and staining combinations Reagents Antibody Fluorochrome Clone Source conjugation CD3 FITC UCHT1 BD Biosciences CD3 PerCP-Cy5.5 SK7 Biolegend

More information

Optimizing Intracellular Flow Cytometry:

Optimizing Intracellular Flow Cytometry: Optimizing Intracellular Flow Cytometry: Simultaneous Detection of Cytokines and Transcription Factors An encore presentation by Jurg Rohrer, PhD, BD Biosciences 10.26.10 Outline Introduction Cytokines

More information

Optimizing Intracellular Flow Cytometry

Optimizing Intracellular Flow Cytometry Optimizing Intracellular Flow Cytometry Detection of Cytokines, Transcription Factors, and Phosphoprotein by Flow Cytometry Presented by Erika O Donnell, PhD, BD Biosciences 23-14876-00 Outline Basic principles

More information

Commercially available HLA Class II tetramers (Beckman Coulter) conjugated to

Commercially available HLA Class II tetramers (Beckman Coulter) conjugated to Class II tetramer staining Commercially available HLA Class II tetramers (Beckman Coulter) conjugated to PE were combined with dominant HIV epitopes (DRB1*0101-DRFYKTLRAEQASQEV, DRB1*0301- PEKEVLVWKFDSRLAFHH,

More information

Supplementary Data. Treg phenotype

Supplementary Data. Treg phenotype Supplementary Data Additional Experiment An additional experiment was performed using cryopreserved peripheral blood mononuclear cells (PBMC) derived from five renal cell carcinoma (RCC) patients [see

More information

Evaluation of Mycobacterium tuberculosis specific T cell response to ESAT-6 and PPD antigen with ELISPOT assay

Evaluation of Mycobacterium tuberculosis specific T cell response to ESAT-6 and PPD antigen with ELISPOT assay In the name of God Evaluation of Mycobacterium tuberculosis specific T cell response to ESAT-6 and PPD antigen with ELISPOT assay presented by: M. Taheri Symptoms of TB Cough for more than two week Bloody

More information

Supplementalgfigureg1gSchematicgdiagramgofgtumor1modellingg

Supplementalgfigureg1gSchematicgdiagramgofgtumor1modellingg SChinjectionh F:LuchLCLsh IVhinjectionh T:cellsh Monitorhforhtumorh growthhandhxeno: reactivehgvhd GVLgexperimentg kcbgvsgpbgt1cellse Xeno1reactiveg experimentg kcbgvsgpbgt1cellse IVhinjectionh 5xh,N^6

More information

Recommended Protocols for Phospho Protein Detection in Human Cells

Recommended Protocols for Phospho Protein Detection in Human Cells Recommended Protocols for Phospho Protein Detection in Human Cells Depending on subcellular localization of the phospho protein of interest, as well as epitope susceptibility to cell fixing and permeabilizing

More information

PBMC from each patient were suspended in AIM V medium (Invitrogen) with 5% human

PBMC from each patient were suspended in AIM V medium (Invitrogen) with 5% human Anti-CD19-CAR transduced T-cell preparation PBMC from each patient were suspended in AIM V medium (Invitrogen) with 5% human AB serum (Gemini) and 300 international units/ml IL-2 (Novartis). T cell proliferation

More information

Table S1. Viral load and CD4 count of HIV-infected patient population

Table S1. Viral load and CD4 count of HIV-infected patient population Table S1. Viral load and CD4 count of HIV-infected patient population Subject ID Viral load (No. of copies per ml of plasma) CD4 count (No. of cells/µl of blood) 28 7, 14 29 7, 23 21 361,99 94 217 7, 11

More information

Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD-

Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD- Supplementary Methods Blocking antibodies and peptides. Rat anti-mouse PD-1 (29F.1A12, rat IgG2a, k), PD- L1 (10F.9G2, rat IgG2b, k), and PD-L2 (3.2, mouse IgG1) have been described (24). Anti-CTLA-4 (clone

More information

Optimizing Intracellular Flow Cytometry:

Optimizing Intracellular Flow Cytometry: Optimizing Intracellular Flow Cytometry: Simultaneous Detection of Cytokines and Transcription Factors Presented by Jurg Rohrer, PhD, BD Biosciences 23-10780-00 Outline Introduction Cytokines Transcription

More information

Fluorochrome Panel 1 Panel 2 Panel 3 Panel 4 Panel 5 CTLA-4 CTLA-4 CD15 CD3 FITC. Bio) PD-1 (MIH4, BD) ICOS (C398.4A, Biolegend) PD-L1 (MIH1, BD)

Fluorochrome Panel 1 Panel 2 Panel 3 Panel 4 Panel 5 CTLA-4 CTLA-4 CD15 CD3 FITC. Bio) PD-1 (MIH4, BD) ICOS (C398.4A, Biolegend) PD-L1 (MIH1, BD) Additional file : Table S. Antibodies used for panel stain to identify peripheral immune cell subsets. Panel : PD- signaling; Panel : CD + T cells, CD + T cells, B cells; Panel : Tregs; Panel :, -T, cdc,

More information

Detailed step-by-step operating procedures for NK cell and CTL degranulation assays

Detailed step-by-step operating procedures for NK cell and CTL degranulation assays Supplemental methods Detailed step-by-step operating procedures for NK cell and CTL degranulation assays Materials PBMC isolated from patients, relatives and healthy donors as control K562 cells (ATCC,

More information

Categorical analysis of human T cell heterogeneity with One-SENSE

Categorical analysis of human T cell heterogeneity with One-SENSE 1 2 3 4 Supplementary Information Categorical analysis of human T cell heterogeneity with One-SENSE 5 6 Running title: T cell Analysis by One-SENSE 7 8 9 1 11 12 13 14 15 16 17 18 19 2 21 22 23 24 25 26

More information

BD Flow Cytometry Reagents Multicolor Panels Designed for Optimal Resolution with the BD LSRFortessa X-20 Cell Analyzer

BD Flow Cytometry Reagents Multicolor Panels Designed for Optimal Resolution with the BD LSRFortessa X-20 Cell Analyzer Multicolor Panels Designed for Optimal Resolution with the BD LSRFortessa X-2 Cell Analyzer Proper multicolor panel design takes into account fluorochrome brightness, antigen density, co-expression, and

More information

Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Willemijn Hobo 1, Wieger Norde 1 and Harry Dolstra 2*

Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Willemijn Hobo 1, Wieger Norde 1 and Harry Dolstra 2* Ex vivo Human Antigen-specific T Cell Proliferation and Degranulation Willemijn Hobo 1, Wieger Norde 1 and Harry Dolstra 2* 1 Department of Laboratory Medicine - Laboratory of Hematology, Radboud University

More information

January 25, 2017 Scientific Research Process Name of Journal: ESPS Manuscript NO: Manuscript Type: Title: Authors: Correspondence to

January 25, 2017 Scientific Research Process Name of Journal: ESPS Manuscript NO: Manuscript Type: Title: Authors: Correspondence to January 25, 2017 Scientific Research Process Name of Journal: World Journal of Gastroenterology ESPS Manuscript NO: 31928 Manuscript Type: ORIGINAL ARTICLE Title: Thiopurine use associated with reduced

More information

SUPPLEMENTARY INFORMATION. Involvement of IL-21 in the epidermal hyperplasia of psoriasis

SUPPLEMENTARY INFORMATION. Involvement of IL-21 in the epidermal hyperplasia of psoriasis SUPPLEMENTARY INFORMATION Involvement of IL-21 in the epidermal hyperplasia of psoriasis Roberta Caruso 1, Elisabetta Botti 2, Massimiliano Sarra 1, Maria Esposito 2, Carmine Stolfi 1, Laura Diluvio 2,

More information

DIFFERENCES IN T-CELL SUBSETS IN ME/CFS. University of Nevada, Reno. Determining the Difference Between Circulating Follicular Regulatory T-cells and

DIFFERENCES IN T-CELL SUBSETS IN ME/CFS. University of Nevada, Reno. Determining the Difference Between Circulating Follicular Regulatory T-cells and DIFFERENCES IN T-CELL SUBSETS IN ME/CFS University of Nevada, Reno Determining the Difference Between Circulating Follicular Regulatory T-cells and Follicular Helper T-cells in Subjects with Myalgic Encephalomyelitis/Chronic

More information

DURACLONE IF BE CERTAIN ABOUT THE RESPONSE. l res. a il n c n. For Research Use Only - Not for use in Diagnostic procedures

DURACLONE IF BE CERTAIN ABOUT THE RESPONSE. l res. a il n c n. For Research Use Only - Not for use in Diagnostic procedures DURACLONE IF earch tria l res lc a om ic il n c n nio pa Yo ur BE CERTAIN ABOUT THE RESPONSE For Research Use Only - Not for use in Diagnostic procedures BE CERTAIN ABOUT THE RESPONSE The sensitive and

More information

MATERIALS AND METHODS. Neutralizing antibodies specific to mouse Dll1, Dll4, J1 and J2 were prepared as described. 1,2 All

MATERIALS AND METHODS. Neutralizing antibodies specific to mouse Dll1, Dll4, J1 and J2 were prepared as described. 1,2 All MATERIALS AND METHODS Antibodies (Abs), flow cytometry analysis and cell lines Neutralizing antibodies specific to mouse Dll1, Dll4, J1 and J2 were prepared as described. 1,2 All other antibodies used

More information

[DOI] /j.issn

[DOI] /j.issn 56 2018 1 1431 - [ ]- - T- SPOT.TB 5638 18T-SPOT.TB T-SPOT.TB86.5%(95%CI 71.2%~95.5%) 100%(95%CI 90.5%~100%) 52.9%(95%CI 27.8%~77.0%) 35.3%(95%CI 14.2%~61.7%) 80.0%(95%CI 64.4%~90.9%) 77.1%(95%CI 62.7%~88.0%)

More information

Challenges in Development and Validation of an Intracellular Cytokine Staining assay

Challenges in Development and Validation of an Intracellular Cytokine Staining assay Challenges in Development and Validation of an Intracellular Cytokine Staining assay Jenny Hendriks, Crucell Hatching @ EBF, Brussels, June 202 www.crucell.com Vaccines vs Protein therapeutics Protein

More information

BD Pharmingen. Human Th1/Th2/Th17 Phenotyping Kit. Technical Data Sheet. Product Information. Description Components:

BD Pharmingen. Human Th1/Th2/Th17 Phenotyping Kit. Technical Data Sheet. Product Information. Description Components: Technical Data Sheet Human Th1/Th2/Th17 Phenotyping Kit Product Information Material Number: Size: Description Components: 560751 50 Tests BD Pharmingen 51-9006615 Human Th1/Th2/Th17 Phenotyping Cocktail

More information

Supplementary Figure 1. IL-12 serum levels and frequency of subsets in FL patients. (A) IL-12

Supplementary Figure 1. IL-12 serum levels and frequency of subsets in FL patients. (A) IL-12 1 Supplementary Data Figure legends Supplementary Figure 1. IL-12 serum levels and frequency of subsets in FL patients. (A) IL-12 serum levels measured by multiplex ELISA (Luminex) in FL patients before

More information

Human and mouse T cell regulation mediated by soluble CD52 interaction with Siglec-10. Esther Bandala-Sanchez, Yuxia Zhang, Simone Reinwald,

Human and mouse T cell regulation mediated by soluble CD52 interaction with Siglec-10. Esther Bandala-Sanchez, Yuxia Zhang, Simone Reinwald, Human and mouse T cell regulation mediated by soluble CD52 interaction with Siglec-1 Esther Bandala-Sanchez, Yuxia Zhang, Simone Reinwald, James A. Dromey, Bo Han Lee, Junyan Qian, Ralph M Böhmer and Leonard

More information

Supplementary Figure 1. Example of gating strategy

Supplementary Figure 1. Example of gating strategy Supplementary Figure 1. Example of gating strategy Legend Supplementary Figure 1: First, gating is performed to include only single cells (singlets) (A) and CD3+ cells (B). After gating on the lymphocyte

More information

Supporting Online Material for

Supporting Online Material for www.sciencemag.org/cgi/content/full/1175194/dc1 Supporting Online Material for A Vital Role for Interleukin-21 in the Control of a Chronic Viral Infection John S. Yi, Ming Du, Allan J. Zajac* *To whom

More information

Supplementary Information

Supplementary Information Supplementary Information Supplementary Figure 1. CD4 + T cell activation and lack of apoptosis after crosslinking with anti-cd3 + anti-cd28 + anti-cd160. (a) Flow cytometry of anti-cd160 (5D.10A11) binding

More information

Primary Adult Naïve CD4+ CD45RA+ Cells. Prepared by: David Randolph at University of Alabama, Birmingham

Primary Adult Naïve CD4+ CD45RA+ Cells. Prepared by: David Randolph at University of Alabama, Birmingham Primary Adult Naïve CD4+ CD45RA+ Cells Prepared by: David Randolph (drdrdr@uab.edu) at University of Alabama, Birmingham Goal: To obtain large numbers of highly pure primary CD4+ CD45RO- CD25- cells from

More information

Supplemental Figure 1. Gating strategies for flow cytometry and intracellular cytokinestaining

Supplemental Figure 1. Gating strategies for flow cytometry and intracellular cytokinestaining Supplemental Figure 1. Gating strategies for flow cytometry and intracellular cytokinestaining of PBMCs. Forward scatter area (FSC-A) versus side scatter area (SSC-A) was used to select lymphocytes followed

More information

BD CBA on the BD Accuri C6: Bringing Multiplexed Cytokine Detection to the Benchtop

BD CBA on the BD Accuri C6: Bringing Multiplexed Cytokine Detection to the Benchtop BD CBA on the BD Accuri C6: Bringing Multiplexed Cytokine Detection to the Benchtop Maria Dinkelmann, PhD Senior Marketing Applications Specialist BD Biosciences, Ann Arbor, MI 23-14380-00 Cellular Communication

More information

APPLICATION OF IMMUNO CHROMATOGRAPHIC METHODS IN PLEURAL TUBERCULOSIS

APPLICATION OF IMMUNO CHROMATOGRAPHIC METHODS IN PLEURAL TUBERCULOSIS APPLICATION OF IMMUNO CHROMATOGRAPHIC METHODS IN PLEURAL TUBERCULOSIS Hadizadeh Tasbiti.AR, Yari.SH, Bahrmand.AR, Karimi.A,Fateh.A, Sayfi.M Tuberculosis Dept.Pasteur Institute of Iran.Tehran.Iran 1 INTRODUCTION

More information

Supplementary Data Table of Contents:

Supplementary Data Table of Contents: Supplementary Data Table of Contents: - Supplementary Methods - Supplementary Figures S1(A-B) - Supplementary Figures S2 (A-B) - Supplementary Figures S3 - Supplementary Figures S4(A-B) - Supplementary

More information

Diagnosis of tuberculosis in children

Diagnosis of tuberculosis in children Diagnosis of tuberculosis in children H Simon Schaaf Desmond Tutu TB Centre Department of Paediatrics and Child Health, Stellenbosch University, and Tygerberg Children s Hospital (TCH) Estimated TB incidence

More information

CD25-PE (BD Biosciences) and labeled with anti-pe-microbeads (Miltenyi Biotec) for depletion of CD25 +

CD25-PE (BD Biosciences) and labeled with anti-pe-microbeads (Miltenyi Biotec) for depletion of CD25 + Supplements Supplemental Materials and Methods Depletion of CD25 + T-cells from PBMC. Fresh or HD precultured PBMC were stained with the conjugate CD25-PE (BD Biosciences) and labeled with anti-pe-microbeads

More information

Supplementary Figures

Supplementary Figures Inhibition of Pulmonary Anti Bacterial Defense by IFN γ During Recovery from Influenza Infection By Keer Sun and Dennis W. Metzger Supplementary Figures d a Ly6G Percentage survival f 1 75 5 1 25 1 5 1

More information

Application Information Bulletin: Human NK Cells Phenotypic characterizing of human Natural Killer (NK) cell populations in peripheral blood

Application Information Bulletin: Human NK Cells Phenotypic characterizing of human Natural Killer (NK) cell populations in peripheral blood Application Information Bulletin: Human NK Cells Phenotypic characterizing of human Natural Killer (NK) cell populations in peripheral blood Christopher A Fraker, Ph.D., University of Miami - Miami, Florida

More information

Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4

Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4 Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4 Ankita Garg, Rodney Trout and Stephen A. Spector,,* Department

More information

<10. IL-1β IL-6 TNF + _ TGF-β + IL-23

<10. IL-1β IL-6 TNF + _ TGF-β + IL-23 3 ns 25 ns 2 IL-17 (pg/ml) 15 1 ns ns 5 IL-1β IL-6 TNF

More information

Online supplement. Phenotypic, functional and plasticity features of classical and alternatively activated

Online supplement. Phenotypic, functional and plasticity features of classical and alternatively activated Online supplement Phenotypic, functional and plasticity features of classical and alternatively activated human macrophages Abdullah Al Tarique*, Jayden Logan *, Emma Thomas *, Patrick G Holt *, Peter

More information

CD14 + S100A9 + Monocytic Myeloid-Derived Suppressor Cells and Their Clinical Relevance in Non-Small Cell Lung Cancer

CD14 + S100A9 + Monocytic Myeloid-Derived Suppressor Cells and Their Clinical Relevance in Non-Small Cell Lung Cancer CD14 + S1A9 + Monocytic Myeloid-Derived Suppressor Cells and Their Clinical Relevance in Non-Small Cell Lung Cancer Po-Hao, Feng M.D., Kang-Yun, Lee, M.D. Ph.D., Ya-Ling Chang, Yao-Fei Chan, Lu- Wei, Kuo,Ting-Yu

More information

Page 1 of 2. Product Information Contents: ezkine Th1 Activation 2 Whole Blood Intracellular Cytokine Kit

Page 1 of 2. Product Information Contents: ezkine Th1 Activation 2 Whole Blood Intracellular Cytokine Kit Page 1 of 2 ezkine Th1 Activation 2 Whole Blood Intracellular Cytokine Kit Catalog Number: 8822-6852 RUO: For Research Use Only. Not for use in diagnostic procedures. Staining of human whole blood with

More information

Supplementary webappendix

Supplementary webappendix Supplementary webappendix This webappendix formed part of the original submission and has been peer reviewed. We post it as supplied by the authors. Supplement to: Portevin D, Moukambi F, Clowes P, et

More information

Interferon γ regulates idiopathic pneumonia syndrome, a. Th17 + CD4 + T-cell-mediated GvH disease

Interferon γ regulates idiopathic pneumonia syndrome, a. Th17 + CD4 + T-cell-mediated GvH disease Interferon γ regulates idiopathic pneumonia syndrome, a Th17 + CD4 + T-cell-mediated GvH disease Nora Mauermann, Julia Burian, Christophe von Garnier, Stefan Dirnhofer, Davide Germano, Christine Schuett,

More information

Supplementary Data 1. Alanine substitutions and position variants of APNCYGNIPL. Applied in

Supplementary Data 1. Alanine substitutions and position variants of APNCYGNIPL. Applied in Supplementary Data 1. Alanine substitutions and position variants of APNCYGNIPL. Applied in Supplementary Fig. 2 Substitution Sequence Position variant Sequence original APNCYGNIPL original APNCYGNIPL

More information

DuraClone IM. Standardized phenotyping panels for studies of the human immune system

DuraClone IM. Standardized phenotyping panels for studies of the human immune system DuraClone IM Standardized phenotyping panels for studies of the human immune system Standardize with the experts, adopt DuraClone IM Tubes. Clinical research studies require accurate, reproducible results

More information

Supplementary Figure 1

Supplementary Figure 1 Supplementary Figure 1 Identification of IFN-γ-producing CD8 + and CD4 + T cells with naive phenotype by alternative gating and sample-processing strategies. a. Contour 5% probability plots show definition

More information

Online Data Supplement. Induction of Pulmonary Granuloma Formation by Propionibacterium acnes is regulated by. MyD88 and Nox2

Online Data Supplement. Induction of Pulmonary Granuloma Formation by Propionibacterium acnes is regulated by. MyD88 and Nox2 Online Data Supplement Induction of Pulmonary Granuloma Formation by Propionibacterium acnes is regulated by MyD88 and Nox2 Jessica L. Werner *, Sylvia G. Escolero *, Jeff T. Hewlett *, Tim N. Mak *3,

More information

CD44

CD44 MR1-5-OP-RU CD24 CD24 CD44 MAIT cells 2.78 11.2 WT RORγt- GFP reporter 1 5 1 4 1 3 2.28 1 5 1 4 1 3 4.8 1.6 8.1 1 5 1 4 1 3 1 5 1 4 1 3 3.7 3.21 8.5 61.7 1 2 1 3 1 4 1 5 TCRβ 2 1 1 3 1 4 1 5 CD44 1 2 GFP

More information

BD Cytofix/Cytoperm Plus Fixation/Permeabilization Kit (with BD GolgiStop protein transport inhibitor containing monensin) (Cat. No.

BD Cytofix/Cytoperm Plus Fixation/Permeabilization Kit (with BD GolgiStop protein transport inhibitor containing monensin) (Cat. No. Kit Manual BD Cytofix/Cytoperm Fixation/Permeabilization Kit (Cat. No. 554714) BD Cytofix/Cytoperm Plus Fixation/Permeabilization Kit (with BD GolgiStop protein transport inhibitor containing monensin)

More information

The CAPRI-T was one of two (along with the CAPRI-NK, see reference. [29]) basic immunological studies nested within the CAMELIA trial.

The CAPRI-T was one of two (along with the CAPRI-NK, see reference. [29]) basic immunological studies nested within the CAMELIA trial. 1 SUPPLEMENTARY INFORMATION Patient description and recruitment The CAPRI-T was one of two (along with the CAPRI-NK, see reference [29]) basic immunological studies nested within the CAMELIA trial. Patients

More information

Induction of granulysin in CD8 T cells by IL-21 and IL-15 is suppressed by human immunodeficiency virus-1

Induction of granulysin in CD8 T cells by IL-21 and IL-15 is suppressed by human immunodeficiency virus-1 Article Induction of granulysin in T cells by and is suppressed by human immunodeficiency virus-1 A. E. Hogg, G. C. Bowick,, N. K. Herzog, M. W. Cloyd,, and J. J. Endsley,,1 Departments of Microbiology

More information

New Tuberculosis Guidelines. Jason Stout, MD, MHS

New Tuberculosis Guidelines. Jason Stout, MD, MHS New Tuberculosis Guidelines Jason Stout, MD, MHS Two New Sets of Guidelines Treatment of Drug-Susceptible Tuberculosis Clinical Infectious Diseases 2016; 63(7): e147-e195 Diagnosis of Tuberculosis in Adults

More information

Recruitment of Mycobacterium tuberculosis specific CD4 + T cells to the site of infection for diagnosis of active tuberculosis

Recruitment of Mycobacterium tuberculosis specific CD4 + T cells to the site of infection for diagnosis of active tuberculosis Original Article doi: 10.1111/j.1365-2796.2008.02012.x Recruitment of Mycobacterium tuberculosis specific CD4 + T cells to the site of infection for diagnosis of active tuberculosis J. Nemeth 1,2, H.-M.

More information

Supplemental Methods. CD107a assay

Supplemental Methods. CD107a assay Supplemental Methods CD107a assay For each T cell culture that was tested, two tubes were prepared. One tube contained BCMA-K562 cells, and the other tube contained NGFR-K562 cells. Both tubes contained

More information

Supplementary Figure 1. Immune profiles of untreated and PD-1 blockade resistant EGFR and Kras mouse lung tumors (a) Total lung weight of untreated

Supplementary Figure 1. Immune profiles of untreated and PD-1 blockade resistant EGFR and Kras mouse lung tumors (a) Total lung weight of untreated 1 Supplementary Figure 1. Immune profiles of untreated and PD-1 blockade resistant EGFR and Kras mouse lung tumors (a) Total lung weight of untreated (U) EGFR TL mice (n=7), Kras mice (n=7), PD-1 blockade

More information

DURACLONE RE THERE ARE EVENTS YOU CANNOT AFFORD TO MISS

DURACLONE RE THERE ARE EVENTS YOU CANNOT AFFORD TO MISS DURACLONE RE THERE ARE EVENTS YOU CANNOT AFFORD TO MISS Your clinical research trial companion For Reseach Use Only - Not for use in Diagnostic procedures THERE ARE EVENTS YOU CANNOT AFFORD TO MISS Rare

More information

Preface. Date: May 1, 2003 Vol.1/MX-2/05/03

Preface. Date: May 1, 2003 Vol.1/MX-2/05/03 Date: May 1, 2003 Vol.1/MX-2/05/03 Preface Tulip Group of companies believes in offering our valued customers the technical support and scientific information to keep updated with the latest international

More information

Title: Role of Interferon-gamma Release Assays in the Diagnosis of Pulmonary Tuberculosis in Patients with Advanced HIV infection

Title: Role of Interferon-gamma Release Assays in the Diagnosis of Pulmonary Tuberculosis in Patients with Advanced HIV infection Author's response to reviews Title: Role of Interferon-gamma Release Assays in the Diagnosis of Pulmonary Tuberculosis in Patients with Advanced HIV infection Authors: Adithya Cattamanchi (acattamanchi@medsfgh.ucsf.edu)

More information

Spleen. mlns. E Spleen 4.1. mlns. Spleen. mlns. Mock 17. Mock CD8 HIV-1 CD38 HLA-DR. Ki67. Spleen. Spleen. mlns. Cheng et al. Fig.

Spleen. mlns. E Spleen 4.1. mlns. Spleen. mlns. Mock 17. Mock CD8 HIV-1 CD38 HLA-DR. Ki67. Spleen. Spleen. mlns. Cheng et al. Fig. C D E F Mock 17 Mock 4.1 CD38 57 CD8 23.7 HLA-DR Ki67 G H I Cheng et al. Fig.S1 Supplementary Figure 1. persistent infection leads to human T cell depletion and hyper-immune activation. Humanized mice

More information

Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis

Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis Immunofluorescent Staining of Intracellular Cytokines for Flow Cytometric Analysis I. Introduction Improved methods have continually been sought to analyze single-cell cytokine production by human and

More information

Supplementary Figure 1. Ex vivo IFNγ production by Tregs. Nature Medicine doi: /nm % CD127. Empty SSC 98.79% CD25 CD45RA.

Supplementary Figure 1. Ex vivo IFNγ production by Tregs. Nature Medicine doi: /nm % CD127. Empty SSC 98.79% CD25 CD45RA. SSC CD25 1.8% CD127 Empty 98.79% FSC CD45RA CD45RA Foxp3 %IFNγ + cells 4 3 2 1 + IL-12 P =.3 IFNγ p=.9 %IL-4+ cells 3 2 1 IL-4 P =.4 c %IL-1 + cells IFNγ 4 3 2 1 Control Foxp3 IL-1 P =.41.64 4.76 MS 2.96

More information

Dan Qiao, 1 Li Li, 1 Jian Guo, 2 Suihua Lao, 3 Xianlan Zhang, 3 Jianping Zhang, 4 and Changyou Wu 1 * on July 7, 2018 by guest

Dan Qiao, 1 Li Li, 1 Jian Guo, 2 Suihua Lao, 3 Xianlan Zhang, 3 Jianping Zhang, 4 and Changyou Wu 1 * on July 7, 2018 by guest INFECTION AND IMMUNITY, Aug. 2011, p. 3358 3365 Vol. 79, No. 8 0019-9567/11/$12.00 doi:10.1128/iai.00014-11 Copyright 2011, American Society for Microbiology. All Rights Reserved. Mycobacterium tuberculosis

More information

Supplementary Figure 1. Enhanced detection of CTLA-4 on the surface of HIV-specific

Supplementary Figure 1. Enhanced detection of CTLA-4 on the surface of HIV-specific SUPPLEMENTARY FIGURE LEGEND Supplementary Figure 1. Enhanced detection of CTLA-4 on the surface of HIV-specific CD4 + T cells correlates with intracellular CTLA-4 levels. (a) Comparative CTLA-4 levels

More information

A step-by-step approach to build and analyze a multicolor panel

A step-by-step approach to build and analyze a multicolor panel Analyze A step-by-step approach to build and analyze a multicolor panel For Research Use Only. Not for use in diagnostic or therapeutic procedures. Alexa Fluor is a registered trademark of Life Technologies

More information

Combined efficacy of pleural fluid lymphocyte neutrophil ratio and pleural fluid adenosine deaminase for the diagnosis of tubercular pleural effusion

Combined efficacy of pleural fluid lymphocyte neutrophil ratio and pleural fluid adenosine deaminase for the diagnosis of tubercular pleural effusion Original article: Combined efficacy of pleural fluid lymphocyte neutrophil ratio and pleural fluid adenosine deaminase for the diagnosis of tubercular pleural effusion Kavita S Kore, Guruprasad Antin,

More information

L-selectin Is Essential for Delivery of Activated CD8 + T Cells to Virus-Infected Organs for Protective Immunity

L-selectin Is Essential for Delivery of Activated CD8 + T Cells to Virus-Infected Organs for Protective Immunity Cell Reports Supplemental Information L-selectin Is Essential for Delivery of Activated CD8 + T Cells to Virus-Infected Organs for Protective Immunity Rebar N. Mohammed, H. Angharad Watson, Miriam Vigar,

More information

Supplementary Fig. 1: Ex vivo tetramer enrichment with anti-c-myc beads

Supplementary Fig. 1: Ex vivo tetramer enrichment with anti-c-myc beads Supplementary Fig. 1: Ex vivo tetramer enrichment with anti-c-myc beads Representative example of comparative ex vivo tetramer enrichment performed in three independent experiments with either conventional

More information

The Diagnosis of Active TB. Deborah McMahan, MD TB Intensive September 28, 2017

The Diagnosis of Active TB. Deborah McMahan, MD TB Intensive September 28, 2017 The Diagnosis of Active TB Deborah McMahan, MD TB Intensive September 28, 2017 Agenda Epidemiology Big picture Conditions that Should Make You Suspicious Which test? Eeenie meenie miny mo Radiographic

More information

HIV-infected subjects were recruited from an ongoing clinic-based cohort (SCOPE)

HIV-infected subjects were recruited from an ongoing clinic-based cohort (SCOPE) SUPPLEMENTAL METHODS Populations studied HIV-infected subjects were recruited from an ongoing clinic-based cohort (SCOPE) based at the University of California, San Francisco. From this cohort, we recruited

More information

Testing for TB. Bart Van Berckelaer Territory Manager Benelux. Subtitle

Testing for TB. Bart Van Berckelaer Territory Manager Benelux. Subtitle Testing for TB Bart Van Berckelaer Territory Manager Benelux Subtitle Agenda TB infection pathway TB immunisation Testing options Pre lab considerations of the whole blood ELISA test The T-SPOT.TB test

More information

Upregulation of Tim-3 and PD-1 expression is associated with tumor antigen specific CD8 + T cell dysfunction in melanoma patients

Upregulation of Tim-3 and PD-1 expression is associated with tumor antigen specific CD8 + T cell dysfunction in melanoma patients Ar ticle Upregulation of Tim-3 and PD-1 expression is associated with tumor antigen specific CD8 + T cell dysfunction in melanoma patients Julien Fourcade, 1 Zhaojun Sun, 1 Mourad Benallaoua, 1 Philippe

More information

Supplementary Materials for

Supplementary Materials for immunology.sciencemag.org/cgi/content/full/2/16/eaan6049/dc1 Supplementary Materials for Enzymatic synthesis of core 2 O-glycans governs the tissue-trafficking potential of memory CD8 + T cells Jossef

More information

Diagnosis and Medical Management of Latent TB Infection

Diagnosis and Medical Management of Latent TB Infection Diagnosis and Medical Management of Latent TB Infection Marsha Majors, RN September 7, 2017 TB Contact Investigation 101 September 6 7, 2017 Little Rock, AR EXCELLENCE EXPERTISE INNOVATION Marsha Majors,

More information

Research Article Citrus/Cydonia Comp. Can Restore the Immunological Balance in Seasonal Allergic Rhinitis-Related Immunological Parameters In Vitro

Research Article Citrus/Cydonia Comp. Can Restore the Immunological Balance in Seasonal Allergic Rhinitis-Related Immunological Parameters In Vitro Hindawi Publishing Corporation Mediators of Inflammation Volume 2008, Article ID 496467, 5 pages doi:10.1155/2008/496467 Research Article Comp. Can Restore the Immunological Balance in Seasonal Allergic

More information

York criteria, 6 RA patients and 10 age- and gender-matched healthy controls (HCs).

York criteria, 6 RA patients and 10 age- and gender-matched healthy controls (HCs). MATERIALS AND METHODS Study population Blood samples were obtained from 15 patients with AS fulfilling the modified New York criteria, 6 RA patients and 10 age- and gender-matched healthy controls (HCs).

More information

ImageStream cytometer analysis. Cells were cultured as described above in vented-cap

ImageStream cytometer analysis. Cells were cultured as described above in vented-cap ImageStream cytometer analysis. Cells were cultured as described above in vented-cap polypropylene tubes, stained with αcd66b-fitc, αm-dc8-pe and αcd56-pe-cy5.5 mabs, washed and fixed with 4 % (w/v) paraformaldehyde.

More information

T-Cell Research. Novel multicolor flow cytometry tools for the study of CD4 + T-cell differentiation and plasticity

T-Cell Research. Novel multicolor flow cytometry tools for the study of CD4 + T-cell differentiation and plasticity T-Cell Research Novel multicolor flow cytometry tools for the study of CD4 + T-cell differentiation and plasticity A Solid Commitment to Research: Flexible Ways to Study CD4 T-Cell Differentiation and

More information

Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained

Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained 1 2 3 4 5 6 7 8 9 10 11 Supplementary Figure 1. H-PGDS deficiency does not affect GI tract functions and anaphylactic reaction. (a) Representative pictures of H&E-stained jejunum sections ( 200 magnification;

More information

Supplementary Figures

Supplementary Figures MiR-29 controls innate and adaptive immune responses against intracellular bacterial infection by targeting IFN-γ Feng Ma 1,2,5, Sheng Xu 1,5, Xingguang Liu 1, Qian Zhang 1, Xiongfei Xu 1, Mofang Liu 3,

More information

Supplemental Methods In vitro T cell assays Inhibition of perforin- and FasL-mediated cytotoxicity Flow Cytometry

Supplemental Methods In vitro T cell assays Inhibition of perforin- and FasL-mediated cytotoxicity Flow Cytometry Supplemental Methods In vitro T cell assays Cell lines Jurkat (ATCC #TIB-152), CCRF-CEM (ATCC #CCL-119), MOLT-4 (ATCC #CRL- 1582), Hut 78 (ATCC #TIB-161), SupT1 (ATCC #CRL-1942), Raji (ATCC #CCL-86) and

More information

Peptide stimulation and Intracellular Cytokine Staining

Peptide stimulation and Intracellular Cytokine Staining v Peptide stimulation and Intracellular Cytokine Staining Authors A. Cosma, S. Allgayer MATERIALS Date 01-02-2007 REAGENTS: Version 1.0 - PBMCs - Culture medium - Costimulating antibodies - Peptide pools

More information

Glow. Save. Let it. and. Get a Discount of 10% BD Biosciences Year End Promotion on a huge Selection of Reagent Solutions

Glow. Save. Let it. and. Get a Discount of 10% BD Biosciences Year End Promotion on a huge Selection of Reagent Solutions Let it Glow and Save BD Biosciences Year End Promotion 2014 Get a Discount of 10% on a huge Selection of Reagent Solutions All fluorochrome labeled antibodies The complete BD CBA portfolio Kits and reagents

More information

Title. CitationCancer science, 109(4): Issue Date Doc URL. Rights(URL)

Title. CitationCancer science, 109(4): Issue Date Doc URL. Rights(URL) Title Toll-like receptor 3 signal augments radiation-induc Yoshida, Sumito; Shime, Hiroaki; Takeda, Yohei; Nam, Author(s) Hiroki; Kasahara, Masanori; Seya, Tsukasa CitationCancer science, 19(): 956-965

More information

For surface staining, cells were stained with various markers (supplemental table S1) at room

For surface staining, cells were stained with various markers (supplemental table S1) at room Supplemental materials and methods. Flow cytometry( details) : For surface staining, cells were stained with various markers (supplemental table S1) at room temperature for 15 minutes, washed with PBS

More information

BMDCs were generated in vitro from bone marrow cells cultured in 10 % RPMI supplemented

BMDCs were generated in vitro from bone marrow cells cultured in 10 % RPMI supplemented Supplemental Materials Figure S1. Cultured BMDCs express CD11c BMDCs were generated in vitro from bone marrow cells cultured in 10 % RPMI supplemented with 15 ng/ml GM-CSF. Media was changed and fresh

More information

Supplementary Information

Supplementary Information Supplementary Information Methods Lymphocyte subsets analysis was performed on samples of 7 subjects by flow cytometry on blood samples collected in ethylenediaminetetraacetic acid (EDTA)-containing tubes

More information

Application Values of T-SPOT.TB in Clinical Rapid Diagnosis of Tuberculosis

Application Values of T-SPOT.TB in Clinical Rapid Diagnosis of Tuberculosis Iran J Public Health, Vol. 47, No.1, Jan 2018, pp.18-23 Original Article Application Values of T-SPOT.TB in Clinical Rapid Diagnosis of Tuberculosis Feng ZHU, Qinfang OU, *Jian ZHENG Dept. of Pulmonology,

More information

Supplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice

Supplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice Supplementary Figure 1. Efficient DC depletion in CD11c.DOG transgenic mice (a) CD11c.DOG transgenic mice (tg) were treated with 8 ng/g body weight (b.w.) diphtheria toxin (DT) i.p. on day -1 and every

More information

B6/COLODR/SPL/11C/83/LAP/#2.006 B6/COLODR/SPL/11C/86/LAP/#2.016 CD11C B6/COLODR/SPL/11C/80/LAP/#2.011 CD11C

B6/COLODR/SPL/11C/83/LAP/#2.006 B6/COLODR/SPL/11C/86/LAP/#2.016 CD11C B6/COLODR/SPL/11C/80/LAP/#2.011 CD11C CD3-specific antibody-induced immune tolerance and suppression of autoimmune encephalomyelitis involves TGF-β production through phagocytes digesting apoptotic T cells Sylvain Perruche 1,3, Pin Zhang 1,

More information

MATERIALS. Peptide stimulation and Intracellular Cytokine Staining- EFFECTOR 45RA PANEL

MATERIALS. Peptide stimulation and Intracellular Cytokine Staining- EFFECTOR 45RA PANEL v Peptide stimulation and Intracellular Cytokine Staining- EFFECTOR 45RA PANEL Authors S. Kutscher, A. Cosma, MATERIALS REAGENTS: - PBMCs - Culture medium - Costimulating antibodies - Peptide pools including

More information

Naive, memory and regulatory T lymphocytes populations analysis

Naive, memory and regulatory T lymphocytes populations analysis Naive, memory and regulatory T lymphocytes populations analysis Jaen Olivier, PhD ojaen@beckmancoulter.com Cellular Analysis application specialist Beckman Coulter France Introduction Flow cytometric analysis

More information

Supplementary Table 1: Summary of Reactogenicity Events by Group and Time of Onset in Entebbe

Supplementary Table 1: Summary of Reactogenicity Events by Group and Time of Onset in Entebbe Supplementary Table 1: Summary of Reactogenicity Events by Group and Time of Onset in Group A (n=39) B (n=11) A (n=39) B (n=11) Onset Post-Vaccination Event 3 Minutes 3-14 days Pain 5 (13%) 1 (1%) 6 (15%)

More information

Mycobacterial Infections: What the Primary Provider Should Know about Tuberculosis

Mycobacterial Infections: What the Primary Provider Should Know about Tuberculosis Mycobacterial Infections: What the Primary Provider Should Know about Tuberculosis Henry F. Chambers, M.D Professor of Medicine, UCSF Topics for Discussion Epidemiology Diagnosis of active TB Screening

More information

Supplemental Information. Human CD1c + Dendritic Cells Drive. the Differentiation of CD103 + CD8 + Mucosal Effector T Cells via the Cytokine TGF-

Supplemental Information. Human CD1c + Dendritic Cells Drive. the Differentiation of CD103 + CD8 + Mucosal Effector T Cells via the Cytokine TGF- Immunity, Volume 38 Supplemental Information Human CD1c + Dendritic Cells Drive the Differentiation of CD103 + CD8 + Mucosal Effector T Cells via the Cytokine TGF- Chun I. Yu Christian Becker Yuanyuan

More information

Abstract. a a a a a

Abstract. a a a a a RESEARCH ARTICLE Production of IgG antibodies to pneumococcal polysaccharides is associated with expansion of ICOS + circulating memory T follicular-helper cells which is impaired by HIV infection a1111111111

More information

Supporting Information

Supporting Information Supporting Information Desnues et al. 10.1073/pnas.1314121111 SI Materials and Methods Mice. Toll-like receptor (TLR)8 / and TLR9 / mice were generated as described previously (1, 2). TLR9 / mice were

More information

Simultaneous correlation of cytokine production with Treg and Th17 cell proliferation

Simultaneous correlation of cytokine production with Treg and Th17 cell proliferation Simultaneous correlation of cytokine production with Treg and Th17 cell proliferation Jurg Rohrer, PhD Director, R&D BD Biosciences 23-11773-00 For Research Use Only. Not for use in diagnostic or therapeutic

More information