What to Measure, How to Measure It

Size: px
Start display at page:

Download "What to Measure, How to Measure It"

Transcription

1 Dale and Betty Bumpers Vaccine Research Center National Institute of Allergy and Infectious Diseases National Institutes of Health Monitoring Memory T-cells: What to Measure, How to Measure It Pratip K. Chattopadhyay, Ph.D. Research Fellow, Roederer Laboratory ImmunoTechnology Section Vaccine Research Center, NIAID, NIH

2 Vaccine Development Choices: antigen, formulation, regimen? How to recognize right choice? Reduced disease incidence/severity = long-term follow-up, or examine early correlates of immunity Efficacy Study Infection Protection Immune Response? Disease time weeks years / decades?

3 Early Correlates of T-cell Immunity What to Measure? Infection Efficacy Study Protection Immune Response? Disease Frequency of antigen-specific T-cells Phenotype Function Aidit Avidity

4 Early Correlates of T-cell Immunity How to Measure Frequency of Ag-specific T-cells? Classical methods: in vitro or ex vivo antigen stimulation, followed by: cell proliferation by tritiated thymidine, or measurement of IFN secretion by ELISA or ELISPOT Considerations: Proliferation assays miss terminal effectors, which do not divide but may still produce cytokines. ELISA and ELISPOT assays measure only IFN Not the only cytokine T-cells produce So, by limiting to IFN, are we missing Ag-specific cells?

5 Measuring the Frequency of Ag-specific T-cells Does IFN Mark All Ag-specific T-cells? 13-color Flow Cytometric Analysis, Acute/Early HIV, n = 471 patients TNFa IL CD IFNg CD CD45RO CD4+ CD8+ 7a CD CD Cells stimulated with overlapping peptides from HIV gagb IFNg CCR7

6 IFN Does Not Mark All Ag-specific T-cells IFN Expression among All Responding* CD8+ Cells 12 9 In this setting, 6% of Ag-specific cells would be missed by ELISpot. 6 IFN - IFN + 3 IFNg (Pies) ( ) + - * defined as a cell expressing at least defined as a cell expressing at least one of the functional markers measured, at levels above the unstimulated control.

7 Measuring the Frequency of Ag-specific T-cells Functional Profile of IFN - Cells 1% CD17a+ IL2- TNF - CD17a- IL2- TNF + Most IFN - cells are CD17a+. Measuring both IFN and CD17a marks 8% of Ag-specific T-cells. CD17a IL2 TNF

8 How to Measure Frequency of Ag-specific T-cells? Proliferation & IFN assays may not mark all Ag-specific cells. Other approaches: Intracellular cytokine staining (more later), including both IFN and CD17a. Tetramers

9 Frequency of Ag-specific T-cells by Tetramer Tetramers structural, not functional, identification Less likely to miss Ag-specific T-cells Antigen-Specific Cells Tetramer+ Cytokine+ Disadvantages: One specificity at a time, not easy for CD4+.

10 Tetramer Staining Considerations How do you know staining is real? 1. Rigorously exclude sources of non-specific binding. dead cells, monocytes, B-cells 2. Practice/optimize staining technique until signal/noise ratio is maximized. Trouble signs: Tetramer+ events in CD4+ population Events positive for more than one tetramer. Background events often naive. Requires combining phenotype + ag-specificity. Reviewed in Chattopadhyay, et al., Cytometry, 28.

11 Early Correlates of T-cell Immunity: Phenotype of Ag-specific T-cells Phenotype often suggests biology: CD57+ cells CD25 br cells CCR7+ cells = short telomeres, terminally differentiated = regulatory T-cells = lymph node homing But how do we choose which phenotypes to measure? Literature: Confusing multiple classification schemes for memory cell Confusing, multiple classification schemes for memory cell subsets are they equivalent?

12 T-cell Classification Schema Compared 13-color expt. Markers Stained: CD3 CD4 Subsets are not equivalent! CD8 CD14 Dead cells (PI) CD45RA CD62L CCR7 CD27 CD28 So how complex is the T-cell compartment? How many markers do we need to measure? CD7 (not shown) CD11a (not shown) CD57 (not shown) Gated on CD8+ T-cells.

13 Complexity of T-cell Compartment: 14-color Flow Di it A / ff t h t > 1% f ll! Diversity: Among memory/effector, no phenotype > 1% of cells! Over 9 phenotypically distinct populations using 6 markers.

14 Are Subsets Really Distinct? Though we find remarkable phenotypic diversity, perhaps some are functionally equivalent, and T-cell compartment not as diverse as it seems. Experiment: Sort various T-cell subsets Stimulate with Cytostim (pan T-cell stimulation) Measure secreted cytokines by Cytokine Bead darray.

15 Phenotypic Diversity Reflects Functional Diversity IFN IL1 IFN IL2 TNF IL2 TNF IFN IL2 TNF Total Cytokine Response Sorted Subset Three CD28+ subsets compared. Some are IL1+ Others do not make IL1. Pheno and function don t always correlate. Two central memory-like populations compared. CD62L+ CCR7+ CD27+ CD28+, IL2 predominates. However, in CD62L+ CD28+ cells TNF predominates. Precise phenotypic definitions important.

16 Phenotypic Correlates of T-cell Immunity: Summary Memory T-cell compartment is diverse. measure more markers : find more distinct subsets Subsets distinct, both in terms of quality and quantity of cytokines. We don t know which to measure, so perhaps better to measure as many as possible?

17 More Colors Help Detect Correlates of Vaccine Efficacy % RA A- X X X X Total CD8+ 1 marker 2 markers 3 markers 4 markers A+B+C+D+ A+B+C+D A+B A B C D A % RA % RA- 57+ X X X X X X X X A = cells expressing a certain combination of proteins (e.g., CD45RA- CD57+ CD27+ CD28+). Bulk measurements include irrelevant populations and reduce our power to detect the cells of interest. RA % X X X X Disease or Vax Efficacy

18 More Colors = More Power the rationale underlying multicolor flow cytometry.

19 More Colors = More Problems? 18-color flow cytometry y described in: Perfetto, Chattopadhyay, et al. Nature Reviews Immunology (24) Chattopadhyay, et al. Nature Medicine (26) Standardization issues: Fluorochrome choice Reagent manufacture Panel design Quality control Data analysis Statistical issues <B515-A>: <G78> <G78-A>: TNFa TNFA 1 4 IFN-G <R71> <R71-A>: CCR7 <R66> <R66-A>: IL2 IL <G56-A>: CD154 Multicolor technology becoming more and more accessible. QuickTime and a decompressor are needed to see this picture.

20 Multicolor Leads to Multiplexing: Qdots Help Organic fluorochromes: 13-color maximum. Broad emission spectra. Overlap = poor staining. Quantum Dots: Inorganic nanocrystals Leap to 18-colors. Narrow emission spectra. Less overlap = sensitive staining Series of Quantum Dots illuminated by violet (45nm) laser.

21 Phenotype + Tetramers : An Example of Multiplexing 17-color experiment: Nature Medicine, August 26 Measured 9 T-cell differentiation markers in four Ag-specific T-cell populations. Qdot-conjugated antibodies to CD45RA and CD57. Qdot pmhci multimers (tetramers) for CMV, EBV, HIV gag, and HIV nef. Data from single tube, one patient sample. Maximum data from minimum sample.

22 Tools to Measure Avidity of Ag-specific T-cells Tetramers : TCR binding stabilized by CD8. Tetramers with mutations in CD8 binding region bind TCR with: less affinity it than wild-type (Choi, et al. JI, 23) = high avidity T-cells, sold to reduce background. more affinity than wild-type (Melenhorst, et al. JIM, 28) = low avidity T-cells May be important in cancer immunology Tumor Ags = self Ags? High affinity tetramers pick up Ag-specific cells that are missed by wild-type tetramers. (Blood, in revision)

23 Early Correlates of T-cell Immunity: Functionality of Ag-Specific T-cells Tetramers: one specificity at a time. difficult for Ag-specific CD4+ T-cells Functional assays: multiple l specificities iti (whole protein, overlapping peptides) can assay CD4 and CD8 responses Recent literature relates simultaneous expression of multiple functions (polyfunctionality) y) to immunity. polyfunctional

24 Summary: Monitoring Memory T-cells may require polychromatic (8+ color) flow cytometry: to enhance the power to detect phenotypes of interest, to improve accuracy, to identify polyfunctional cells. However, T-cell monitoring: doesn t always have to be done in one tube, may only require live purification of Ag-specific cells, simplified by markers that correlate with complex function.

25 Simple Purification of Live Ag-specific T-cells Live = can t use ICS, fix/perm is lethal CD8+ Tetramers or CD17a (6-7% of Ag-specific T-cells) CD4+ CD154 upon stimulation, use to isolate Ag-specific cells? Problem: transient expression, rapidly internalized. Solution: CD154 in culture during stim + monensin (Chattopadhyay, et al. Nature Medicine, 25.)

26 Kinetics of CD154 Expression Time point = 5 1 Time point = 1 hours 5 1 Time point = 2 5 Time point = 3 hours 1 2 hours 3 hours d4 <Cblue-A>: c d4 <Cblue-A>: c d4 <Cblue-A>: c d4 <Cblue-A>: c % %.3% 2.5% <PE-A>: cd154 keepers Event 1Count: Time point = 4 hours 4 hours <PE-A>: cd154 keepers 1 Event 1Count: Time point = 5 hours <PE-A>: cd154 keepers 2 Event 1Count: Time point = 6 hours <PE-A>: cd154 keepers 3 Event 1Count: Time point = 12 hours 5 hours 6 hours 8 hours <Cblue-A>: cd d <Cblue-A>: cd d <Cblue-A>: cd d <Cblue-A>: cd d % 6.3% 14% 16% 21% <PE-A>: cd154 keepers 4 Event Count: Time point = 18 hours <Cblue-A>: cd4 CD4 12 hours <PE-A>: cd154 keepers 6 Event Count: hours Time point = 24 hours keepers 18 Event Count: <PE-A>: cd154 CD <Cblue-A>: cd % 23% keepers 24 Event Count: <PE-A>: cd <PE-A>: cd154 keepers 8 Event Count: <PE-A>: cd154 keepers 12 Event Count: Hours

27 Cytokine Expressing Cells Are CD <FIT TC-A>: IFN FNG <PE E-A>: CD % % % 91.6% <Ax68-A>: IL2 IL <Cy7PE-A>: TNF TNFA % of IL2+ or IFN + cells express CD % of these cells also express TNF. Most cytokine+ cells express CD154.

28 Not All CD154+ Cells Are IFN IL2, or TNFa+ TNF Negative TNF and IL2 and IFN and negative CD4 T IL2 Negative and 5.69 IFN Negative Boolean Gating CD154 Assay identifies responding cells that make IL4, IL5, and IL1 thirty hours later.

29 Isolation of Live, Highly Cytotoxic Cells CD17a = not specific for cytotoxic activity. Any stimulated CD8+ cell will degranulate, regardless of endosomal content. Cytotoxic enzymes (CEs): granzyme A, granzyme B, perforin CE combination matters: knockouts of each more susceptible to infection than wildtype. Immunity may require cooperative action. Can we identify cells with all three CEs, without fix/perm?

30 Isolation of Live, Highly Cytotoxic Cells CD57 bright marks grza+ grzb+ perforin high cells. Means to identify these cells without fix/perm, and only one marker. CD57 delineates function in a way common classification schemes don t. Chattopadhyay, et al. Journal Leukocyte Biology, in press.

31 Summary Many ways to monitor human memory T-cells, each with special considerations. To quantify Ag-specific cells: IFN alone is not enough! Tetramers least likely to miss responses but limited specificities, not easy for CD4 Functional assays biologically relevant, assay multiple epitopes which function? polychromatic flow cytometry Live cell assays for Ag-specific cells CD4+ : CD154 assay CD8+ : CD17a assay

32 Summary Is quantity important, or does cell quality matter? Phenotypic analysis can be combined with tetramer or functional assays, and checks quality of tetramer staining. Note that memory cell compartment very diverse, cells identified with one marker set not equivalent to others. Functional analysis polyfunctional cells imp. in many successful immune responses. live cell purification: highly cytotoxic cells expressing CD57. Avidity another potential way of assessing quality tools emerging, relevance not fully known yet.

33 Acknowledgements David Price Joseph Melenhorst, AJ Barrett Mario Roederer Steve Perfetto Joanne Yu Tess Brodie Yolanda Mahnke Carl-Magnus Hogerkorp Diane Bolton Enrico Lugli Kaimei Song Steve De Rosa Mike Betts Rick Koup Danny Douek Bob Seder

34

35 Highly Cytotoxic Cells Found in All Subsets N CM CM Naïve Central Mem Memory Effector E CM E CM M A- B- P- A+ B- P- A+ B+ P- A+ B+ P+ grza+ grzb+ perforin high found in many memory subsets. CD57 delineates function in a way common classification schemes don t. With only one marker! Not seen before polychromatic flow.

36 CD154 : Marker for CD4+ T-cell Responses? Only expressed upon stimulation, restricted to CD Fresh PBMC Cblue-A> >: CD4 1 4 CD4 < % <PE-A>: CD154 CD hours SEB stimulation Surface staining CD3, CD4, CD8, CD154 Gated on CD3+ lymphocytes. H i ll l l CD4+ ll d IFN However, in parallel sample, more CD4+ cells expressed IFN So, surface staining cannot identify all Ag-responsive CD4+ s.

37 Improving CD154 Measurements Problem: Transient expression. Surface CD154 6 hours Solution: Stain with CD154 PE during stimulation. Detect CD154 recycled from cell surface. Use monensin. Once internalized, complex may degrade in acidic endosome. Monensin prevents endosomal acidification. So, it s added d to prevent PE degradation d during long staining. i

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

Estimating primate effector T cell responses to DNA vaccination

Estimating primate effector T cell responses to DNA vaccination Estimating primate effector T cell responses to DNA vaccination Oct 22 nd, 21 Devon J. Shedlock, PhD th Vaccine Renaissance Conference Estimating vaccineinduced effector T cell responses Vaccineinduced

More information

HD1 (FLU) HD2 (EBV) HD2 (FLU)

HD1 (FLU) HD2 (EBV) HD2 (FLU) ramer staining + anti-pe beads ramer staining a HD1 (FLU) HD2 (EBV) HD2 (FLU).73.11.56.46.24 1.12 b CD127 + c CD127 + d CD127 - e CD127 - PD1 - PD1 + PD1 + PD1-1 1 1 1 %CD127 + PD1-8 6 4 2 + anti-pe %CD127

More information

Cytotoxicity assays. Rory D. de Vries, PhD 1. Viroscience lab, Erasmus MC, Rotterdam, the Netherlands

Cytotoxicity assays. Rory D. de Vries, PhD 1. Viroscience lab, Erasmus MC, Rotterdam, the Netherlands Cytotoxicity assays Rory D. de Vries, PhD 1 1 Viroscience lab, Erasmus MC, Rotterdam, the Netherlands Anti-influenza immunity Humoral / CD4+ / CD8+ / NK? Function of CTL Elimination of virus-infected cells?

More information

Rapid perforin upregulation directly ex vivo by CD8 + T cells is a defining characteristic of HIV elite controllers

Rapid perforin upregulation directly ex vivo by CD8 + T cells is a defining characteristic of HIV elite controllers Rapid perforin upregulation directly ex vivo by CD8 + T cells is a defining characteristic of HIV elite controllers Adam R. Hersperger Department of Microbiology University of Pennsylvania Evidence for

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

Bead Based Assays for Cytokine Detection

Bead Based Assays for Cytokine Detection Bead Based Assays for Cytokine Detection September 27, 2014 6 th EFIS-EJI South East European Immunology School SEEIS 2014 Timisoara, Romania The Cells of the Immune System The Immune Reaction (Th2) (Th1)

More information

Why are validated immunogenicity assays important for HIV vaccine development?

Why are validated immunogenicity assays important for HIV vaccine development? Why are validated immunogenicity assays important for HIV vaccine development? There is a need to compare immunogenicity of products in the pipeline, when similar or different in class when developed by

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

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 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

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

Vaccination in Humans Generates Broad T Cell Cytokine Responses

Vaccination in Humans Generates Broad T Cell Cytokine Responses This information is current as of November 2, 2018. Vaccination in Humans Generates Broad T Cell Cytokine Responses Stephen C. De Rosa, Fabien X. Lu, Joanne Yu, Stephen P. Perfetto, Judith Falloon, Susan

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

Low Avidity CMV + T Cells accumulate in Old Humans

Low Avidity CMV + T Cells accumulate in Old Humans Supplementary Figure Legends Supplementary Figure 1. CD45RA expressing CMVpp65-specific T cell populations accumulate within HLA-A*0201 and HLA-B*0701 individuals Pooled data showing the size of the NLV/HLA-A*0201-specific

More information

System Biology analysis of innate and adaptive immune responses during HIV infection

System Biology analysis of innate and adaptive immune responses during HIV infection System Biology analysis of innate and adaptive immune responses during HIV infection Model of T cell memory persistence and exhaustion Naive Ag+APC Effector TEM (Pfp, Gr.B, FasL, TNF) Ag stim. IL-2, IL-7,

More information

Vaccine Delivery and TLR Ligands Influence the Quality of T cell Responses in NHP. Robert A. Seder, M.D. Vaccine Research Center NIAID, NIH

Vaccine Delivery and TLR Ligands Influence the Quality of T cell Responses in NHP. Robert A. Seder, M.D. Vaccine Research Center NIAID, NIH Vaccine Delivery and TLR Ligands Influence the Quality of T cell Responses in NHP Robert A. Seder, M.D. Vaccine Research Center NIAID, NIH Vaccines Against HIV, Malaria and Tuberculosis Will Require Neutralizing

More information

How to set up a good protocol or Polychromatic flow cytometry: Advantages and pitfalls

How to set up a good protocol or Polychromatic flow cytometry: Advantages and pitfalls Universität Leipzig How to set up a good protocol or Polychromatic flow cytometry: Advantages and pitfalls Attila Tarnok UNIVERSITÄT LEIPZIG H E R Z Z E N T R U M Dept. of Pediatric Cardiology, Heart Center,

More information

Innate and Cellular Immunology Control of Infection by Cell-mediated Immunity

Innate and Cellular Immunology Control of Infection by Cell-mediated Immunity Innate & adaptive Immunity Innate and Cellular Immunology Control of Infection by Cell-mediated Immunity Helen Horton PhD Seattle Biomedical Research Institute Depts of Global Health & Medicine, UW Cellular

More information

Supplemental materials

Supplemental materials Supplemental materials 1 Supplemental Fig. 1 Immunogram This immunogram summarizes patient clinical data and immune parameters at corresponding time points for Patient IMF-32. The top panel illustrates

More information

Identification and Characterization of CD4 T cells actively transcribing HIV RNA in Peripheral Blood

Identification and Characterization of CD4 T cells actively transcribing HIV RNA in Peripheral Blood Dale and Betty Bumpers Vaccine Research Center National Institute of Allergy and Infectious Diseases National Institutes of Health Identification and Characterization of CD4 T cells actively transcribing

More information

Micro 204. Cytotoxic T Lymphocytes (CTL) Lewis Lanier

Micro 204. Cytotoxic T Lymphocytes (CTL) Lewis Lanier Micro 204 Cytotoxic T Lymphocytes (CTL) Lewis Lanier Lewis.Lanier@ucsf.edu Lymphocyte-mediated Cytotoxicity CD8 + αβ-tcr + T cells CD4 + αβ-tcr + T cells γδ-tcr + T cells Natural Killer cells CD8 + αβ-tcr

More information

MHC Tetramers and Monomers for Immuno-Oncology and Autoimmunity Drug Discovery

MHC Tetramers and Monomers for Immuno-Oncology and Autoimmunity Drug Discovery MHC Tetramers and Monomers for Immuno-Oncology and Autoimmunity Drug Discovery Your Partner in Drug Discovery and Research MHC Tetramer Background T-Cell Receptors recognize and bind to complexes composed

More information

TCR, MHC and coreceptors

TCR, MHC and coreceptors Cooperation In Immune Responses Antigen processing how peptides get into MHC Antigen processing involves the intracellular proteolytic generation of MHC binding proteins Protein antigens may be processed

More information

MHC MULTIMER PROFICIENCY PANEL 2017

MHC MULTIMER PROFICIENCY PANEL 2017 MHC MULTIMER PROFICIENCY PANEL 2017 August 2017 CONTACT Charlotte Halgreen ProficiencyPanel@immudex.com FOR MORE INFORMATION www.proficiencypanel.com MHC MULTIMER PROFICIENCY PANEL 2017 This report summarizes

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

Engineering an in vitro human immune system for rapid vaccine evaluation

Engineering an in vitro human immune system for rapid vaccine evaluation Engineering an in vitro human immune system for rapid vaccine evaluation William Warren, Ph.D. VaxDesign Corporation Orlando, FL 32826 wwarren@vaxdesign.com www.vaxdesign.com 1 MIMIC Technology Overview

More information

NK mediated Antibody Dependent Cellular Cytotoxicity in HIV infections

NK mediated Antibody Dependent Cellular Cytotoxicity in HIV infections NK mediated Antibody Dependent Cellular Cytotoxicity in HIV infections Amy Chung Dr. Ivan Stratov Prof. Stephen Kent ADCC process consists of Target cell QuickTime and a TIFF (Uncompressed) FcγR decompressor

More information

Cellular Immunity in Aging and HIV: Correlates of Protection. Immune Senescence

Cellular Immunity in Aging and HIV: Correlates of Protection. Immune Senescence Cellular Immunity in Aging and HIV: Correlates of Protection Janet E. McElhaney, MD Professor of Medicine Allan M. McGavin Chair in Research Geriatrics University of British Columbia Vancouver, BC and

More information

Perforin Expression Directly Ex Vivo by HIV-Specific CD8+ T-Cells Is. Is a Correlate of HIV Elite Control

Perforin Expression Directly Ex Vivo by HIV-Specific CD8+ T-Cells Is. Is a Correlate of HIV Elite Control Perforin Expression Directly Ex Vivo by HIV-Specific CD8+ T-Cells Is a Correlate of HIV Elite Control The Harvard community has made this article openly available. Please share how this access benefits

More information

Superior T memory stem cell persistence supports long-lived T cell memory

Superior T memory stem cell persistence supports long-lived T cell memory Brief report Superior T memory stem cell persistence supports long-lived T cell memory Enrico Lugli, 1,2 Maria H. Dominguez, 1 Luca Gattinoni, 3,4 Pratip K. Chattopadhyay, 1 Diane L. Bolton, 1 Kaimei Song,

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

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

Journal of Immunological Methods

Journal of Immunological Methods Journal of Immunological Methods 338 (2008) 31 39 Contents lists available at ScienceDirect Journal of Immunological Methods journal homepage: www.elsevier.com/locate/jim Research paper Detection of low

More information

The central role of T helper cells

The central role of T helper cells e central role of T helper s the expanding universe of lineages Cytometric analysis of T function Hyun-Dong Chang Deutsches Rheuma-Forschungszentrum Berlin, a Leibniz Institute, Berlin, Germany CD8+ T

More information

x Lymphocyte count /µl CD8+ count/µl 800 Calculated

x Lymphocyte count /µl CD8+ count/µl 800 Calculated % Lymphocyte in CBC A. 50 40 30 20 10 Lymphocyte count /µl B. x10 3 2.5 1.5 C. 50 D. 1000 % CD3+CD8+ Cells 40 30 20 Calculated CD8+ count/µl 800 600 400 200 10 0 #61 #63 #64 #65 #68 #71 #72 #75 Figure

More information

IMMUNOLOGICAL MEMORY. CD4 T Follicular Helper Cells. Memory CD8 T Cell Differentiation

IMMUNOLOGICAL MEMORY. CD4 T Follicular Helper Cells. Memory CD8 T Cell Differentiation IMMUNOLOGICAL MEMORY CD4 T Follicular Helper Cells Memory CD8 T Cell Differentiation CD4 T Cell Differentiation Bcl-6 T-bet GATA-3 ROR t Foxp3 CD4 T follicular helper (Tfh) cells FUNCTION Provide essential

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

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

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

staining and flow cytometry

staining and flow cytometry Detection of influenza virus-specific T cell responses by intracellular by cytokine intracellular staining cytokine staining and flow cytometry Detection of influenza virus-specific T cell responses and

More information

B and T-Cells. CD8+ Cytotoxic T Cells

B and T-Cells. CD8+ Cytotoxic T Cells Sony Biotechnology Inc. Application Data, SA38 Spectral Analyzer TBNK Panel Page 1 TBNK panel on the SA38 Spectral Analyzer TBNK (B-Cell, T-Cell, and natural killer cells) panels are frequently used to

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

Potential cross reactions between HIV 1 specific T cells and the microbiome. Andrew McMichael Suzanne Campion

Potential cross reactions between HIV 1 specific T cells and the microbiome. Andrew McMichael Suzanne Campion Potential cross reactions between HIV 1 specific T cells and the microbiome Andrew McMichael Suzanne Campion Role of the Microbiome? T cell (and B cell) immune responses to HIV and Vaccines are influenced

More information

Cellular Immune response. Jianzhong Chen, Ph.D Institute of immunology, ZJU

Cellular Immune response. Jianzhong Chen, Ph.D Institute of immunology, ZJU Cellular Immune response Jianzhong Chen, Ph.D Institute of immunology, ZJU Concept of adaptive immune response T cell-mediated adaptive immune response I. Concept of immune response A collective and coordinated

More information

T cell-mediated immunity

T cell-mediated immunity T cell-mediated immunity Overview For microbes within phagosomes in phagocytes.cd4+ T lymphocytes (TH1) Activate phagocyte by cytokines studies on Listeria monocytogenes For microbes infecting and replicating

More information

Supplementary Figure 1. Using DNA barcode-labeled MHC multimers to generate TCR fingerprints

Supplementary Figure 1. Using DNA barcode-labeled MHC multimers to generate TCR fingerprints Supplementary Figure 1 Using DNA barcode-labeled MHC multimers to generate TCR fingerprints (a) Schematic overview of the workflow behind a TCR fingerprint. Each peptide position of the original peptide

More information

CD4 + T-cell-inducing HIV vaccines may have an impact on viral control

CD4 + T-cell-inducing HIV vaccines may have an impact on viral control CD4 + T-cell-inducing HIV vaccines may have an impact on viral control Clues from cytokines produced by CD4 + T-cells from HIVinfected patients and vaccinated seronegative volunteers Eva Van Braeckel 1,

More information

CD4 + CD25 high Foxp3 + T regulatory cells kill autologous CD8(+) and CD4(+) T cells using Fas/FasL- and Granzyme B- mediated pathways

CD4 + CD25 high Foxp3 + T regulatory cells kill autologous CD8(+) and CD4(+) T cells using Fas/FasL- and Granzyme B- mediated pathways CD4 + CD25 high Foxp3 + T regulatory cells kill autologous CD8(+) and CD4(+) T cells using Fas/FasL- and Granzyme B- mediated pathways Laura Strauss, Christoph Bergmann, Theresa L. Whiteside University

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

Therapeutic PD L1 and LAG 3 blockade rapidly clears established blood stage Plasmodium infection

Therapeutic PD L1 and LAG 3 blockade rapidly clears established blood stage Plasmodium infection Supplementary Information Therapeutic PD L1 and LAG 3 blockade rapidly clears established blood stage Plasmodium infection Noah S. Butler, Jacqueline Moebius, Lecia L. Pewe, Boubacar Traore, Ogobara K.

More information

Immune response. This overview figure summarizes simply how our body responds to foreign molecules that enter to it.

Immune response. This overview figure summarizes simply how our body responds to foreign molecules that enter to it. Immune response This overview figure summarizes simply how our body responds to foreign molecules that enter to it. It s highly recommended to watch Dr Najeeb s lecture that s titled T Helper cells and

More information

Biology of Immune Aging

Biology of Immune Aging Biology of Immune Aging Jorg J. Goronzy Stanford University Immune deficiency Increase morbidity and mortality from infections Poor vaccine responses Cancer Immune Aging Chronic inflammation Coronary artery

More information

Long-term persistence of T cell memory in Italian vaccinees

Long-term persistence of T cell memory in Italian vaccinees Long-term persistence of T cell memory in Italian vaccinees Mario (Mago) Clerici Head, Department of Medical Sciences and Biotechnologies Head, Doctorate School in Molecular Medicine University of Milano

More information

Supplementary Figure 1. mrna expression of chitinase and chitinase-like protein in splenic immune cells. Each splenic immune cell population was

Supplementary Figure 1. mrna expression of chitinase and chitinase-like protein in splenic immune cells. Each splenic immune cell population was Supplementary Figure 1. mrna expression of chitinase and chitinase-like protein in splenic immune cells. Each splenic immune cell population was sorted by FACS. Surface markers for sorting were CD11c +

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

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

RAISON D ETRE OF THE IMMUNE SYSTEM:

RAISON D ETRE OF THE IMMUNE SYSTEM: RAISON D ETRE OF THE IMMUNE SYSTEM: To Distinguish Self from Non-Self Thereby Protecting Us From Our Hostile Environment. Innate Immunity Acquired Immunity Innate immunity: (Antigen nonspecific) defense

More information

Clinical Translation of Immunotherapy using WT1 and CMV specific TCR Gene Transfer

Clinical Translation of Immunotherapy using WT1 and CMV specific TCR Gene Transfer Clinical Translation of Immunotherapy using WT1 and CMV specific Gene Transfer Dr Emma C Morris Reader, Dept of Immunology, UCL Consultant Haematologist (BMT), UCLH and RFH ISCT, 2/5/211 Gene Transfer

More information

Cellular biomarkers of T lymphocyte func6on. Michael Be9s University of Pennsylvania Department of Microbiology

Cellular biomarkers of T lymphocyte func6on. Michael Be9s University of Pennsylvania Department of Microbiology Cellular biomarkers of T lymphocyte func6on Michael Be9s University of Pennsylvania Department of Microbiology What T cell response parameters could be important for an effec6ve T cell based product? Specificity

More information

Supplementary Figure 1. Characterization of basophils after reconstitution of SCID mice

Supplementary Figure 1. Characterization of basophils after reconstitution of SCID mice Supplementary figure legends Supplementary Figure 1. Characterization of after reconstitution of SCID mice with CD4 + CD62L + T cells. (A-C) SCID mice (n = 6 / group) were reconstituted with 2 x 1 6 CD4

More information

Scott Abrams, Ph.D. Professor of Oncology, x4375 Kuby Immunology SEVENTH EDITION

Scott Abrams, Ph.D. Professor of Oncology, x4375 Kuby Immunology SEVENTH EDITION Scott Abrams, Ph.D. Professor of Oncology, x4375 scott.abrams@roswellpark.org Kuby Immunology SEVENTH EDITION CHAPTER 13 Effector Responses: Cell- and Antibody-Mediated Immunity Copyright 2013 by W. H.

More information

Nine-Color Flow Cytometry for Accurate Measurement of T Cell Subsets and Cytokine Responses. Part I: Panel Design by an Empiric Approach

Nine-Color Flow Cytometry for Accurate Measurement of T Cell Subsets and Cytokine Responses. Part I: Panel Design by an Empiric Approach Original Article Nine-Color Flow Cytometry for Accurate Measurement of T Cell Subsets and Cytokine Responses. Part I: Panel Design by an Empiric Approach Bridget E. McLaughlin, 1 Nicole Baumgarth, 2 Martin

More information

From the Diagnostic Immunology Laboratories

From the Diagnostic Immunology Laboratories Immunology Laboratories ISSUE 9 SPRING 2012 PAGE 1 Spring Meetings Please stop by and say hello in May at this year s American Society of Pediatric Hematology/Oncology (ASPHO) Annual Meeting in New Orleans

More information

Adaptive (acquired) immunity. Professor Peter Delves University College London

Adaptive (acquired) immunity. Professor Peter Delves University College London Adaptive (acquired) immunity Professor Peter Delves University College London p.delves@ucl.ac.uk Haematopoiesis Haematopoiesis Lymphocytes = adaptive response Recognition of pathogens by adaptive cells,

More information

Avidity for antigen shapes clonal dominance in CD8 T cell populations specific for persistent DNA viruses

Avidity for antigen shapes clonal dominance in CD8 T cell populations specific for persistent DNA viruses Avidity for antigen shapes clonal dominance in CD8 T cell populations specific for persistent DNA viruses David A. Price, 1 Jason M. Brenchley, 1 Laura E. Ruff, 1 Michael R. Betts, 2 Brenna J. Hill, 1

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

T Memory Stem Cells: A Long-term Reservoir for HIV-1

T Memory Stem Cells: A Long-term Reservoir for HIV-1 Towards an HIV Cure Pre-Conference Symposium 20 & 21 July 2012 T Memory Stem Cells: A Long-term Reservoir for HIV-1 Maria J Buzon, PhD Ragon Institute of MGH, MIT and Harvard, Massachussetts General Hospital,

More information

Antigen Presentation and T Lymphocyte Activation. Abul K. Abbas UCSF. FOCiS

Antigen Presentation and T Lymphocyte Activation. Abul K. Abbas UCSF. FOCiS 1 Antigen Presentation and T Lymphocyte Activation Abul K. Abbas UCSF FOCiS 2 Lecture outline Dendritic cells and antigen presentation The role of the MHC T cell activation Costimulation, the B7:CD28 family

More information

Normally occurring NKG2D + CD4 + T cells are immunosuppressive and inversely correlated with disease activity in juvenileonset

Normally occurring NKG2D + CD4 + T cells are immunosuppressive and inversely correlated with disease activity in juvenileonset Normally occurring NKG2D + CD4 + T cells are immunosuppressive and inversely correlated with disease activity in juvenileonset lupus Zhenpeng Dai, 1 Cameron J. Turtle, 1 Garrett C. Booth, 1 Stanley R.

More information

Supporting Information

Supporting Information Supporting Information Chapuis et al. 10.1073/pnas.1113748109 SI Methods Selection of Patients, Targets, Isolation, and Expansion of Melanoma- Specific CTL Clones. Patients were HLA-typed, and their tumors

More information

Chapter 3, Part A (Pages 37-45): Leukocyte Migration into Tissues

Chapter 3, Part A (Pages 37-45): Leukocyte Migration into Tissues Allergy and Immunology Review Corner: Chapter 3, Part A (pages 37-45) of Cellular and Molecular Immunology (Seventh Edition), by Abul K. Abbas, Andrew H. Lichtman and Shiv Pillai. Chapter 3, Part A (Pages

More information

T cell memory Friday, February 10, 17

T cell memory Friday, February 10, 17 T cell memory 2-10-2017 expansion contraction memory recall EFFECTORS 10 6 pathogen load secondary challenge MEMORY 10 2 NAIVE 2 7-8 30 >1 year days post-infection In mouse, ~100 naïve, CD8s become ~3x10

More information

UNISEC Universal Influenza Vaccines Secured

UNISEC Universal Influenza Vaccines Secured UNISEC Universal Influenza Vaccines Secured Partners in the UNISEC Consortium UNISEC has been made possible by contributions from the European Commission DG-Research and the European member states Project

More information

Alessandra Franco, M.D., Ph.D. UCSD School of Medicine Department of Pediatrics Division of Allergy, Immunology and Rheumatology

Alessandra Franco, M.D., Ph.D. UCSD School of Medicine Department of Pediatrics Division of Allergy, Immunology and Rheumatology Natural regulatory T cells recognize the heavy constant region (Fc) of immunoglobulins: a novel mechanism for IVIG immunotherapy in Pediatric Immune-mediated diseases Alessandra Franco, M.D., Ph.D. UCSD

More information

Immune response to infection

Immune response to infection Immune response to infection Dr. Sandra Nitsche (Sandra.Nitsche@rub.de ) 20.06.2018 1 Course of acute infection Typical acute infection that is cleared by an adaptive immune reaction 1. invasion of pathogen

More information

Medical Virology Immunology. Dr. Sameer Naji, MB, BCh, PhD (UK) Head of Basic Medical Sciences Dept. Faculty of Medicine The Hashemite University

Medical Virology Immunology. Dr. Sameer Naji, MB, BCh, PhD (UK) Head of Basic Medical Sciences Dept. Faculty of Medicine The Hashemite University Medical Virology Immunology Dr. Sameer Naji, MB, BCh, PhD (UK) Head of Basic Medical Sciences Dept. Faculty of Medicine The Hashemite University Human blood cells Phases of immune responses Microbe Naïve

More information

Effect of Cytomegalovirus Infection on Immune Responsiveness. Rene van Lier Sanquin Blood Supply Foundation

Effect of Cytomegalovirus Infection on Immune Responsiveness. Rene van Lier Sanquin Blood Supply Foundation Effect of Cytomegalovirus Infection on Immune Responsiveness Rene van Lier Sanquin Blood Supply Foundation OUTLINE CMV infection and vaccination responses Effects of CMV infection on CD8 + T cell numbers

More information

Parallel session IVB. Chair: Hans Wildiers

Parallel session IVB. Chair: Hans Wildiers Parallel session IVB Biology of ageing and cancer Chair: Hans Wildiers IMMUNOSENESCENCE AND AGEING Tamas Fulop M.D., PhD Research Center on Aging, Division of Geriatrics, University of Sherbrooke, Sherbrooke,

More information

Normally occurring NKG2D + CD4 + T cells are immunosuppressive and inversely correlated with disease activity in juvenileonset

Normally occurring NKG2D + CD4 + T cells are immunosuppressive and inversely correlated with disease activity in juvenileonset ARTICLE Normally occurring NKG2D + CD4 + T cells are immunosuppressive and inversely correlated with disease activity in juvenileonset lupus Zhenpeng Dai, 1 Cameron J. Turtle, 1 Garrett C. Booth, 1 Stanley

More information

NKTR-255: Accessing The Immunotherapeutic Potential Of IL-15 for NK Cell Therapies

NKTR-255: Accessing The Immunotherapeutic Potential Of IL-15 for NK Cell Therapies NKTR-255: Accessing The Immunotherapeutic Potential Of IL-15 for NK Cell Therapies Saul Kivimäe Senior Scientist, Research Biology Nektar Therapeutics NK Cell-Based Cancer Immunotherapy, September 26-27,

More information

IFN-γ Secretion Assay Detection Kit (PE) human

IFN-γ Secretion Assay Detection Kit (PE) human Miltenyi Biotec GmbH Friedrich-Ebert-Straße 68 51429 Bergisch Gladbach, Germany Phone +49 2204 8306-0 Fax +49 2204 85197 macs@miltenyibiotec.de Miltenyi Biotec Inc. 12740 Earhart Avenue Auburn CA 95602,

More information

Cancer Biometrics: Results of the 2003 isbtc Workshop. Theresa L. Whiteside, Ph.D., ABMLI University of Pittsburgh Cancer Institute

Cancer Biometrics: Results of the 2003 isbtc Workshop. Theresa L. Whiteside, Ph.D., ABMLI University of Pittsburgh Cancer Institute Cancer Biometrics: Results of the 2003 isbtc Workshop Theresa L. Whiteside, Ph.D., ABMLI University of Pittsburgh Cancer Institute The workshop objective The objective was to consider state-of-theart approaches

More information

Immunology Basics Relevant to Cancer Immunotherapy: T Cell Activation, Costimulation, and Effector T Cells

Immunology Basics Relevant to Cancer Immunotherapy: T Cell Activation, Costimulation, and Effector T Cells Immunology Basics Relevant to Cancer Immunotherapy: T Cell Activation, Costimulation, and Effector T Cells Andrew H. Lichtman, M.D. Ph.D. Department of Pathology Brigham and Women s Hospital and Harvard

More information

Adaptive Immunity. Jeffrey K. Actor, Ph.D. MSB 2.214,

Adaptive Immunity. Jeffrey K. Actor, Ph.D. MSB 2.214, Adaptive Immunity Jeffrey K. Actor, Ph.D. MSB 2.214, 500-5344 Lecture Objectives: Understand role of various molecules including cytokines, chemokines, costimulatory and adhesion molecules in the development

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

MHC MULTIMER PROFICIENCY PANEL 2015

MHC MULTIMER PROFICIENCY PANEL 2015 MHC MULTIMER PROFICIENCY PANEL 2015 July 2015 CONTACT Charlotte Halgreen ProficiencyPanel@immudex.com FOR MORE INFORMATION www.proficiencypanel.com MHC MULTIMER PROFICIENCY PANEL 2015 This report summarizes

More information

NK cell flow cytometric assay In vivo DC viability and migration assay

NK cell flow cytometric assay In vivo DC viability and migration assay NK cell flow cytometric assay 6 NK cells were purified, by negative selection with the NK Cell Isolation Kit (Miltenyi iotec), from spleen and lymph nodes of 6 RAG1KO mice, injected the day before with

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

CONTRACTING ORGANIZATION: Johns Hopkins University School of Medicine Baltimore, MD 21205

CONTRACTING ORGANIZATION: Johns Hopkins University School of Medicine Baltimore, MD 21205 AD Award Number: DAMD7---7 TITLE: Development of Artificial Antigen Presenting Cells for Prostate Cancer Immunotherapy PRINCIPAL INVESTIGATOR: Jonathan P. Schneck, M.D., Ph.D. Mathias Oelke, Ph.D. CONTRACTING

More information

Dissecting T cell activation and function at the single cell level

Dissecting T cell activation and function at the single cell level Dissecting T cell activation and function at the single cell level Regulation and outcome of immune cell interactions in vivo Lymph node In vivo imaging of immune responses with two-photon microscopy

More information

General Overview of Immunology. Kimberly S. Schluns, Ph.D. Associate Professor Department of Immunology UT MD Anderson Cancer Center

General Overview of Immunology. Kimberly S. Schluns, Ph.D. Associate Professor Department of Immunology UT MD Anderson Cancer Center General Overview of Immunology Kimberly S. Schluns, Ph.D. Associate Professor Department of Immunology UT MD Anderson Cancer Center Objectives Describe differences between innate and adaptive immune responses

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

Supplementary Figure 1. Normal T lymphocyte populations in Dapk -/- mice. (a) Normal thymic development in Dapk -/- mice. Thymocytes from WT and Dapk

Supplementary Figure 1. Normal T lymphocyte populations in Dapk -/- mice. (a) Normal thymic development in Dapk -/- mice. Thymocytes from WT and Dapk Supplementary Figure 1. Normal T lymphocyte populations in Dapk -/- mice. (a) Normal thymic development in Dapk -/- mice. Thymocytes from WT and Dapk -/- mice were stained for expression of CD4 and CD8.

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

Immune response and correlates of protection against typhoid

Immune response and correlates of protection against typhoid Immune response and correlates of protection against typhoid Marcelo B. Sztein, M.D. Immunology Group Center for Vaccine Development University of Maryland Correlates of enteric vaccine-induced protection

More information

Nature Medicine: doi: /nm.2109

Nature Medicine: doi: /nm.2109 HIV 1 Infects Multipotent Progenitor Cells Causing Cell Death and Establishing Latent Cellular Reservoirs Christoph C. Carter, Adewunmi Onafuwa Nuga, Lucy A. M c Namara, James Riddell IV, Dale Bixby, Michael

More information

Nature Immunology: doi: /ni Supplementary Figure 1. Examples of staining for each antibody used for the mass cytometry analysis.

Nature Immunology: doi: /ni Supplementary Figure 1. Examples of staining for each antibody used for the mass cytometry analysis. Supplementary Figure 1 Examples of staining for each antibody used for the mass cytometry analysis. To illustrate the functionality of each antibody probe, representative plots illustrating the expected

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

Adaptive Immunity: Humoral Immune Responses

Adaptive Immunity: Humoral Immune Responses MICR2209 Adaptive Immunity: Humoral Immune Responses Dr Allison Imrie 1 Synopsis: In this lecture we will review the different mechanisms which constitute the humoral immune response, and examine the antibody

More information