ndln NK Cells (x10 3 ) Days post-infection (A/PR/8) *** *** *** Liver NK Cells (x10 4 ) Days post-infection (MCMV)

Similar documents
Licensing delineates helper and effector NK cell subsets during viral infection

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

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

% of live splenocytes. STAT5 deletion. (open shapes) % ROSA + % floxed

SUPPLEMENTARY INFORMATION

Supporting 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.

ILC1 and ILC3 isolation and culture Following cell sorting, we confirmed that the recovered cells belonged to the ILC1, ILC2 and

Supplementary Figure 1 Cytokine receptors on developing thymocytes that can potentially signal Runx3d expression.

Supplemental Figure 1

Supplementary Figure 1. Example of gating strategy

Supporting Information

SUPPLEMENTARY FIGURES

a surface permeabilized

Supplementary Figures

Combined Rho-kinase inhibition and immunogenic cell death triggers and propagates immunity against cancer

Supplementary information. The proton-sensing G protein-coupled receptor T-cell death-associated gene 8

Supplementary Figure 1

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

SUPPLEMENTARY FIGURE 1

Supplemental Information. Lung gd T Cells Mediate Protective Responses. during Neonatal Influenza Infection. that Are Associated with Type 2 Immunity

Supplemental Table I.

well for 2 h at rt. Each dot represents an individual mouse and bar is the mean ±

Supplementary Figures

Cytokine therapy reverses NK cell anergy in MHC-deficient tumors

Supplementary Figures

Immunity and Cancer. Doriana Fruci. Lab di Immuno-Oncologia

Supplementary Figure 1. Generation of knockin mice expressing L-selectinN138G. (a) Schematics of the Sellg allele (top), the targeting vector, the

pplementary Figur Supplementary Figure 1. a.

Supplementary Table 1

The encephalitogenicity of TH17 cells is dependent on IL-1- and IL-23- induced production of the cytokine GM-CSF

Krishnamoorthy et al.,

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

Supplemental Figure 1. Signature gene expression in in vitro differentiated Th0, Th1, Th2, Th17 and Treg cells. (A) Naïve CD4 + T cells were cultured

Hua Tang, Weiping Cao, Sudhir Pai Kasturi, Rajesh Ravindran, Helder I Nakaya, Kousik

SUPPLEMENTARY INFORMATION

Supplementary Figure 1. Deletion of Smad3 prevents B16F10 melanoma invasion and metastasis in a mouse s.c. tumor model.

Defective STAT1 activation associated with impaired IFN-g production in NK and T lymphocytes from metastatic melanoma patients treated with IL-2

Supplementary information CD4 T cells are required for both development and maintenance of disease in a new model of reversible colitis

Supplementary Figure 1. NAFL enhanced immunity of other vaccines (a) An over-the-counter, hand-held non-ablative fractional laser (NAFL).

Supplementary Fig. 1 p38 MAPK negatively regulates DC differentiation. (a) Western blot analysis of p38 isoform expression in BM cells, immature DCs

SUPPLEMENTARY METHODS

Supplementary Figures

Supplementary Materials for

Dendritic cell subsets and CD4 T cell immunity in Melanoma. Ben Wylie 1 st year PhD Candidate

Generation of ST2-GFP reporter mice and characterization of ILC1 cells following infection

<20 <20 <20 < <20 <20 <20 <20. Mock

Treatment with IL-7 Prevents the Decline of Circulating CD4 + T Cells during the Acute Phase of SIV Infection in Rhesus Macaques

days days and gbt-i.cd Recipient 20

Supplemental Materials for. Effects of sphingosine-1-phosphate receptor 1 phosphorylation in response to. FTY720 during neuroinflammation

B220 CD4 CD8. Figure 1. Confocal Image of Sensitized HLN. Representative image of a sensitized HLN

Supplemental Table 1. Primer sequences for transcript analysis

SUPPLEMENTARY INFORMATION

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

Supplementary Figure 1. BMS enhances human T cell activation in vitro in a

Supplementary Figure 1 Chemokine and chemokine receptor expression during muscle regeneration (a) Analysis of CR3CR1 mrna expression by real time-pcr

Supplementary Information

Online Appendix Material and Methods: Pancreatic RNA isolation and quantitative real-time (q)rt-pcr. Mice were fasted overnight and killed 1 hour (h)

Control GST GST-RAP. α2-mg. 170 kda. b-actin. 42 kda LRP-1

Nature Medicine: doi: /nm.2109

Eosinophils are required. for the maintenance of plasma cells in the bone marrow

Tbk1-TKO! DN cells (%)! 15! 10!

Supplementary Figure 1.

Supplementary Materials for

Supplemental Materials

SUPPORTING INFORMATIONS

IL-34 is a tissue-restricted ligand of CSF1R required for the development of Langerhans cells and microglia

IOM Immunization Safety Review 11/12/2001. Immunological Competition and the Infant Immune Response to Vaccines

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

Supplemental Information. Aryl Hydrocarbon Receptor Controls. Monocyte Differentiation. into Dendritic Cells versus Macrophages

Supporting Information

NMED-A65251A. Supplementary Figures.

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

Supplementary Information:

Live cell imaging of trafficking of the chaperone complex vaccine to the ER. BMDCs were incubated with ER-Tracker Red (1 M) in staining solution for

Title: NATURAL KILLER CELL FUNCTIONS AND SURFACE RECEPTORS

Supplementary Figure 1. Efficiency of Mll4 deletion and its effect on T cell populations in the periphery. Nature Immunology: doi: /ni.

Nature Medicine: doi: /nm.3922

ECM1 controls T H 2 cell egress from lymph nodes through re-expression of S1P 1

SUPPLEMENTARY INFORMATION

CD4 + T cells recovered in Rag2 / recipient ( 10 5 ) Heart Lung Pancreas

Supplementary Information. Tissue-wide immunity against Leishmania. through collective production of nitric oxide

Supporting Online Material for

Supporting Information Table of Contents

Supplementary Figure 1: Fn14 is upregulated in the epidermis and dermis of mice

Supplemental Information. The Therapeutic Effect. of Anti-HER2/neu Antibody Depends. on Both Innate and Adaptive Immunity CONTENTS:

Reviewers' comments: Reviewer #1 (Remarks to the Author):

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

Supplementary Figures for TSC1 controls macrophage polarization to prevent inflammatory disorder by Linnan Zhu et al

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

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

Endogenous TNFα orchestrates the trafficking of neutrophils into and within lymphatic vessels during acute inflammation

SUPPLEMENTARY INFORMATION

Supporting Information

Supplementary Figure 1

Supporting Information

ACTIVATION AND EFFECTOR FUNCTIONS OF CELL-MEDIATED IMMUNITY AND NK CELLS. Choompone Sakonwasun, MD (Hons), FRCPT

L1 on PyMT tumor cells but Py117 cells are more responsive to IFN-γ. (A) Flow

Supplemental materials

Nature Immunology: doi: /ni Supplementary Figure 1. Cytokine pattern in skin in response to urushiol.

Transcription:

A mln NK Cells(x ) 6 1 * ndln NK Cells (x ) ns C Lung NK Cells(x ) 1 1 7 * D LN NK Cells (x ) 1 7 1 7 Days post-infection (A/PR/8) * * E Liver NK Cells (x ) 1 7 Days post-infection (A/PR/8) * * * 1 7 Days post-infection (A/PR/8) 7 1 7 1 Days post-infection (MCMV) Days post-infection (MCMV) Supplementary Figure 1. Trafficking of NK cells to priming and target tissues throughout the course of viral infection. (A-E) Time course of absolute numbers of NK cells (CD-NK1.1+) in the mln) (A), ndln) (), lung (C), draining lymph nodes (DLN) (D), and liver (E), of C7L/6 at different time points post-apr8 (A-C) or post-mcmv (D,E) infection. Control (white-filled bars) mice were mock-infected by receiving either μl of PS (APR8 control) i.n. or. ml of RPMI (MCMV control) i.p., whereas infected mice (blackfilled bars) were either infected with PFU APR8 in μl of PS i.n. or x PFU MCMV in. ml of RPMI i.p. n= or mice per group representative of - experiments. One-way ANOVA with Tukey post-test used to compare groups. * p<., p<.1, * p<.1.

A CD7-CD11b- CD7+CD11b- CD7+CD11b+ CD7-CD11b+ * 1 / / / / CD7-CD11b- CD7+CD11b- CD7+CD11b+ CD7-CD11b+ * * NK Cell subsets (%) Control mln (H b ) A/PR/8 # of NK Cells (x ) * * * * Control A/PR/8 Supplementary Figure. Trafficking of NK cells to priming and target tissues throughout the course of viral infection. (A) Frequency of immature (CD7-CD11b-, CD7+CD11b-) and mature (CD7+CD11b+, CD7-CD11b+) NK cells in the mediastinal lymph nodes (mln) of C7L/6 (Hb) mice on day post-apr8 infection. () Absolute number of immature and mature unlicensed and licensed NK cell subsets in the mln of C7L/6 (Hb) mice on day post-apr8 infection. n= mice per group representative of - experiments. Two-way ANOVA with Tukey post-test used to compare groups. p<.1, *p<.1.

K K K K 1K 1K K K Singlets 8. K FSC-H K 1K K K Lymphocytes 78. K Ly9C/I 7.1 K K K FSC-A 1K K CD K FSC-A NK Cells.87 K NK1.1 Ly9G 9.1 K Ly9C/I A K K K 1K 1K 1K K K K K K K.6 Ly9C/I Ly9G Ly9A Ly9A 8. 1.. - Ly9G. 17.1 Ly9A.1 K. 6.9 Ly9G oolean Exclusion Gating Ly9 Negative Single positive Ly9s Ly9A-C/I-G.7 Double positive Ly9s Ly9A-C/I+G17. K K Triple positive Ly9s Ly9A-C/I+G+ 1.9 Ly9A+C/I+G+. 1K Ly9C/I Ly9C/I Ly9A+C/I-G. K Ly9C/I K Ly9C/I Ly9G K Ly9G Ly9G Ly9A+C/I-G+ 9.6 7.1% K 1K Ly9A Ly9A-C/I-G+.8 K Ly9G 6.9% Ly9A+C/I+G.67 K 1K - Ly9C/I+G+A+ (.%) Ly9C/I+G+A- (1.9%) Ly9C/I+G-A+ (.67%) Ly9C/I+G-A- (17.%) K Ly9A K Ly9C/I-G+A+ (9.6%) Ly9C/I-G+A- (.8%) Ly9C/I-G-A+ (.%) Ly9C/I-G-A- (.7%) K Ly9A K - Ly9G Receptor inds to Ly9C/I Hb Ly9G Hd Ly9A Hd C7L/6 (Hb).D (Hd) Ly9G Receptor inds to C7L/6 (Hb).D (Hd) Ly9C/I+G+A+ Hb, Hd Ly9C/I+G+A- H, H d Ly9C/I+G-A+ H, H d Ly9C/I+G-ALy9C/I-G+A+ b H b H d Ly9C/I-G+A- Hd Ly9C/I-G-A+ H Ly9C/I-G-A- b - d Supplementary Figure. Comparison between boolean gating and conventional gating to determine the phenotype of NK cell subsets. (A) Representative gating strategy comparing boolean gating (red square) and conventional gating (grey square) to identify Ly9-receptor subsets by staining for Ly9A, -G, and C/I on splenocytes from C7L/6 mice. oolean gating (red square) allows for the simultaneous identification of Ly9-negative, single-positive, double-positive, and triple-positive Ly9-expressing NK cell subsets, whereas conventional gating allows for pairwise comparisons of Ly9 receptors. oolean gating allows for the identification and quanitification of CD-NK1.1+Ly9C/I+G+A+,

Supplementary Figure continued. Ly9C/I+G+A-, Ly9C/I+G-A+, Ly9C/I+G-A-, Ly9C/I-G+A+, Ly9C/I-G+A-, Ly9C/I-G-A+, and Ly9C/I-G-A- NK cell subsets, represented as percentages of each subset among total NK cells in the pie chart (bottom). () Classification of licensed and unlicensed NK cell subsets in C7L/6 (Hb) and.d (Hd) mice based on the ability of specified Ly9 receptors to bind MHC class I molecules using either conventional gating (grey square) or oolean gating (red square).

A 6% (±%) Unl A/PR/8 mln (H d ) Ly9C/I+G+A+ (1.%) Ly9C/I+G+A- (9.%) Ly9C/I+G-A+ (.1%) Ly9C/I-G+A+ (.%) Ly9C/I-G+A- (16.8%) Ly9C/I-G-A+ (.9%) Ly9C/I+G-A- (1.9%) Ly9C/I-G-A- (.%) % (±%) NK Cells (x ) 1 9 6 Control A/PR/8 mln (H d ) * C 7% (±%) Unl A/PR/8 Lung (H d ) Ly9C/I+G+A+ (1.7%) Ly9C/I+G+A- (1.7%) Ly9C/I+G-A+ (6.%) Ly9C/I-G+A+ (7.%) Ly9C/I-G+A- (.%) Ly9C/I-G-A+ (11.%) Ly9C/I+G-A- (1.7%) Ly9C/I-G-A- (.%) 6% (±%) D NK Cells (x ) 1 Control A/PR/8 Lung (H d ) * * Supplementary Figure..D NK subsets show similar licensing distribution patterns following viral infection. (A) Frequency or () absolute number of licensed (CD-NK1.1+Ly9C/I+G+A+, Ly9C/I+G+A-, Ly9C/I+G-A+, Ly9C/I-G+A+, Ly9C/I-G+A-, or Ly9C/I-G-A+; shades of red) and unlicensed (CD-NK1.1+Ly9C/I+G-A-, or Ly9C/I-G-A-; shades of blue) NK cell subsets in the mediastinal lymph nodes (mln) of.d (Hd) mice on day post-apr8 infection. (C) Frequency or (D) absolute number of licensed (shades of red) and unlicensed (shades of blue) NK cell subsets in the lung of.d (Hd) mice on day post-apr8 infection. n= mice per group representative of - experiments. (,D) Two-way ANOVA with Tukey post-test used to compare groups.

Lung CD1 D Liver C Lymph Nodes CD CD CD A CD1 CD1 E F Ly9C/I Ly9I 7.7 9.71 H 1.6.9 Ly9A.1.17-1.7 1. 8. Ly9G 9. 17.8 6.6 9..7 -.7 8.9.7 Ly9G. 7. - I - 8.1 Ly9A Ly9A - Ly9G Ly9G Ly9A Ly9I Ly9G G Ly9I Ly9G.1.1-9.8 9.9 Ly9G Supplementary Figure. Verification of successful depletion of specific NK populations with in vivo monoclonal antibody depletion in C7L/6 mice. (A-C) non-depleted migg (left) and NK1.1 (PK16) (right) depleted C7L/6 mice had lungs (A), livers (), and lymph nodes (C) stained for CD and CD1. (D-F) Mice were either administered mouse/rat IgG (top) or given either anti-ly9c/i (E6) (middle) or anti-ly9g (D11) (bottom) and lungs (D), livers (E), and lymph nodes (F) were removed and stained for CD-NK1.1+ and Ly9G (clone Cwy) vs Ly9I (clone YLI-9). (G-I) Mice were either administered rat IgG (top) or given anti-ly9a (YE1/) (bottom) and lungs (G), livers (H), and lymph nodes (I) were removed and stained for CD-NK1.1+ and Ly9G (clone D11) vs Ly9A (clone YE1/8..6). µg of rigg, anti-nk1.1, anti-ly9g, anti-ly9c/i, or anti-ly9a in. ml of PS i.p. was given days prior to infections. Flow plots are representative of mice from separate experiments.

A Liver CD1 CD1 Lymph Nodes CD1 E D C CD CD CD Lung F Ly9C/I Ly9I Ly9I Ly9I Ly9G Ly9G Ly9G Ly9G Supplementary Figure 6: Verification of successful depletion of specific NK populations with in vivo monoclonal antibody depletion in.d mice. (A-C) non-depleted migg (left) and NK1.1 (PK16) (right) depleted.d mice had lungs (A), livers (), and lymph nodes (C) stained for CD and CD1. (D-F) Mice were either administered mouse/rat IgG (top) or given either anti-ly9c/i (E6) (middle) or anti-ly9g (D11) (bottom) and lungs (D), livers (E), and lymph nodes (F) were removed and stained for CD-NK1.1+ and Ly9G (clone Cwy) vs Ly9I (clone YLI-9). µg of rigg, anti-nk1.1, anti-ly9g, or anti-ly9c/i in. ml of PS I.P. was given days prior to infections. Flow plots are representative of mice from separate experiments.

A Ly9C/I+ Ly9C/I+ Ly9A+ Ly9A+ C D GM-CSF+ NK Cells (x ) Ly9G+ * * mln (H d ) - A/PR8 IFNγ+ NK Cells (x ) 1 Ly9G+ * Lung (H d ) - A/PR8 Ratio of GM-CSF to IFNγ.6... * % of GM-CSF+ NK Cells 6 1 NKGA+ ns NKGA- Supplementary Figure 7..D and human NK subsets display cytokine profiles consistent with licensing. (A) Absolute number of NK cells that are GM-CSF+ and either CD-NK1.1+Ly9G+, Ly9A+, or Ly9C/I+ at day after APR8 infection in the mediastinal lymph nodes (mln) of.d mice. () Absolute number of NK cells that are IFN-γ+ and either CD-NK1.1+Ly9G+, Ly9A+, or Ly9C/I+ at day after APR8 infection in the lung of.d. (C) Aggregate ratio of GM-CSF to IFN-γ production by unlicensed and licensed NK cells. (D) Percentage of NKGA+ or NKGA- among either licensed or unlicensed NK cell populations that are GM-CSF+ after 16 hours of PMA/Ionomycin stimulation. Murine data: n= mice per group representative of - experiments. One-way ANOVA with Tukey posttest used to compare groups. Human data: Experiments were performed in duplicates; 1- aggregated single-positive NK cell subsets per unlicensed or licensed group. Five distinct individuals were used to obtain NK cells from PMCs. * p<., p<.1, * p<.1.

2024 © healthdocbox.com Privacy Policy | Terms of Service | Feedback