Excerpt from MACS&more Vol 17 1/2016. Kenji Miki¹, Koji Nagaoka¹, Hermann Bohnenkamp², Takayuki Yoshimoto³, Ryuji Maekawa¹*, and Takashi Kamigaki⁴

Size: px
Start display at page:

Download "Excerpt from MACS&more Vol 17 1/2016. Kenji Miki¹, Koji Nagaoka¹, Hermann Bohnenkamp², Takayuki Yoshimoto³, Ryuji Maekawa¹*, and Takashi Kamigaki⁴"

Transcription

1 Excerpt from MCS&more Vol 17 1/2016 Dendritic cells pulsed with induce both OV-specific CD4 + and CD8 + T cells and cause antitumor effects in a mouse model of lymphoma Kenji Miki¹, Koji Nagaoka¹, Hermann ohnenkamp², Takayuki Yoshimoto³, Ryuji Maekawa¹, and Takashi Kamigaki⁴ ¹ Medinet Medical Ititute, MEDINET Co., Ltd., Tokyo, Japan; ² Miltenyi iotec GmbH, ergisch Gladbach, Germany; ³ Ititute of Medical Science, Tokyo Medical University, Tokyo, Japan; ⁴ Seta Clinic Group, Shin-Yokohama, Japan Miltenyi iotec provides products and services worldwide. Visit to find your nearest Miltenyi iotec contact. Unless otherwise specifically indicated, Miltenyi iotec products and services are for research use only and not for therapeutic or diagnostic use. MCS, the MCS logo, and are registered trademarks or trademarks of Miltenyi iotec GmbH. Copyright 2016 Miltenyi iotec GmbH. ll rights reserved.

2 Dendritic cells pulsed with induce both OV-specific CD4 + and CD8 + T cells and cause antitumor effects in a mouse model of lymphoma Kenji Miki¹, Koji Nagaoka¹, Hermann ohnenkamp², Takayuki Yoshimoto³, Ryuji Maekawa¹, and Takashi Kamigaki⁴ ¹ Medinet Medical Ititute, MEDINET Co., Ltd., Tokyo, Japan ² Miltenyi iotec GmbH, ergisch Gladbach, Germany ³ Ititute of Medical Science, Tokyo Medical University, Tokyo, Japan ⁴ Seta Clinic Group, Shin-Yokohama, Japan not shown Introduction Dendritic cell (DC)-based vaccines hold great promise for cancer immunotherapy. DCs are pulsed with peptides and subsequently used as antigen-presenting cells to induce an antitumor respoe in vivo through the activation of T cells. Thus far, mostly epitope-specific peptides with 8 10 amino acids in length have been used to generate DC vaccines, which however activate only CD8 + T cells. Moreover, available single epitope-specific peptides are restricted to MHCI or MHCII and therefore activate either CD8 + or CD4 + T cells. Here we used a Pool, which coists mainly of 15-mer peptides covering the complete sequence of the target antigen, in this case ovalbumin (OV), with an 11 amino acid overlap. We show that DCs pulsed with this peptide pool induced both OV-specific CD8 + and CD4 + T cell respoes and caused strong antitumor effects in a mouse model of lymphoma. Material and methods DC generation and antigen loading DCs were generated from mouse (C57L/6) bone marrow cells cultured in the presence of 20 ng/ml GM-CSF for 10 days. Subsequently, DCs were maturated in the presence of 10 ng/ ml GM-CSF, 10 ng/ml IL-4, and 1 µg/ml LPS. DCs were loaded with antigen by pulsing with peptide pool (Miltenyi iotec) for 4 hours. (OV) peptides with I-b (MHCII)-restricted (OV ) and H-2Kb (MHCI)-restricted (OV ) epitopes were used as positive controls. ll peptides were used at the final concentration of 2 µg/ml. Following the incubation, cells were washed with medium to remove excess peptides. E.G7 DCs T cell isolation T cells were obtained from splee of OT-II or OT-I mice carrying a tragenic OVspecific T cell receptor (TCR). CD4 + and CD8 + T cells were isolated with the CD4 + T Cell Isolation Kit, mouse and the CD8a + T Cell Isolation Kit, mouse, respectively (both from Miltenyi iotec). T cell priming capacity of DCs To test the capacity of DCs to induce T cell proliferation, the DCs pulsed with or the I-b- or H-2Kb-restricted OV peptides were cocultured with CFSElabeled or. The ratio of DCs to T cells was 1:20. Untreated DCs were used as a negative control. fter 1 to 4 days of coculture, T cell proliferation was determined by flow cytometry using absolute (I- b ) (H-2K b ) (I- b + H-2K b ) OV tetramer assay Serum IgG detection by ELIS Measurement of tumor weight C57L/ Figure 1 Timeline for the evaluation of antitumor effects of DCs. Numbers indicate the days after tumor induction with E.G7 cells. Vol 17 1/2016 MCS & more 7

3 count beads. Moreover, proliferation of T cells was analyzed on day 3 by a CFSE dilution assay. On day 2, concentratio of IL-2 and IFN-γ in the culture supernatants were measured by ELIS. ssessment of antitumor effects of DCs in a mouse model of lymphoma The timeline for tumor induction, immunization with DCs, and evaluation of antitumor effects by DCs is outlined in figure 1. Tumors were induced in C57L/6 mice on day 0 by subcutaneously injecting 5 10⁵ E.G7-OV tumor cells. E.G7-OV cells are derivatives of EL4 mouse lymphoma cells modified to express and secrete OV cotitutively. Therefore, these cells can be recognized by pulsed DCs. fter 4 days, the mice were injected with 3 10⁵ DCs that were either left untreated or pulsed with or MHCII (I-b)- restricted or MHCI (H-2Kb)-restricted OV peptides or both peptides in combination. The tumor volume was measured with a micrometer calliper at various time points until day 14. On day 14, six mice were sacrificed to measure tumor size by weight. Evaluation of DC-induced immune respoes in a mouse model On day 11, serum from three mice was examined for the presence of OV-specific IgG by ELIS. Moreover, the numbers of OVspecific CD8 + cytotoxic T cells were determined by flow cytometry. To this end, resected tumors were dissociated into single-cell suspeio by treatment with collagenase. Subsequently, cells were analyzed by flow cytometry for H-2K b OV tetramer + CD8 + T cells. Cell numbers were normalized to the weight of tumors to allow for direct comparison of the individual mice. Results OV peptide pulsed DCs induce CD4 + and CD8 + T cells to proliferate and secrete pro-inflammatory cytokines Mature DCs have the capacity to induce T cell proliferation. To test whether the OV peptide pulsed DCs can exert this function, we cocultured the DCs with CD4 + or CD8 + T cells carrying an OV-specific TCR and measured the numbers of T cells after various time points. C IL-2 level (ng/ml) IFN-γ level (ng/ml) T cell number ( 10⁵) Relative cell number Time (days) no antigen peptide (I- b ) no antigen peptide (I- b ) 10⁰ 10¹ 10² 10³ CFSE (I- b ) (I- b ) IL-2 IFN-γ Figure 2 Capacity of DCs to induce T cell proliferation and secretion of proinflammatory cytokines. DCs loaded with or the MHCII (I- b ) or MHCI (H-2Kb) restricted OV peptides were cocultured with isolated CD4 + or CD8 + T cells on day 0. T cell numbers were determined by flow cytometry at various time points (; n = 3; mea±sd; Student s t-test; : non-significant at p 5). On day 3, T cell proliferation was assessed by flow cytometric CFSE dilution assay (). Concentratio of IL-2 and IFN-γ in the supernatant of the coculture were determined on day 2 (C; n = 3; mea±sd; Student s t-test; : non-significant at p 5). T cell number ( 10⁵) IL-2 level (ng/ml) IFN-γ level (ng/ml) Relative cell number ⁰ 10¹ 10² 10³ CFSE Time (days) no antigen peptide (H-2K b ) no antigen peptide (H-2K b ) (H-2K b ) (H-2K b ) 8 MCS & more Vol 17 1/2016

4 3000 Tumor volume (mm³) volume with all pulsed DCs. DCs pulsed with or the two MHCIand MHCII-restricted OV peptides led to the largest reduction in tumor size by about 90%, followed by the MHCI-restricted peptide (80% reduction). The inhibitory effect of DCs pulsed with the MHCII-restricted peptide was the smallest, leading to a reduction in tumor size by about 50% (fig. 3). Similar results were obtained for the tumor weight ten days after immunization (fig. 3). Tumor weight (mg) Time after tumor inoculation (days) (I- b ) (H-2K b ) (I- b + H-2K b ) (I- b ) (H-2K b ) (I- b + H-2K b ) Figure 3 Inhibition of tumor growth by OV-pulsed DCs. C57L/6 mice were injected with E.G7 cells on day 0. () Tumor volume was measured with a micrometer calliper at various time points after immunization with DCs that were loaded with or the MHCII (I-b) and/or MHCI (H-2Kb) restricted OV peptides. () t day 14 after injection of tumor cells, i.e., day 10 after DC immunization, mice were sacrificed and the tumor size was measured in weight. ( and ; n = 6; mea±sd; Student s t-test; : non-significant at p 5; p < 5. DCs loaded with induced an increase in the numbers of both CD4 + and CD8 + T cells from day 2 onward, similarly to the respective MHCII or MHCIrestricted peptides. DCs that were not loaded with any antigen did not induce T cell proliferation (fig. 2). T cell proliferation was confirmed with the CFSE dilution assay on day 3 (fig. 2). Likewise, DCs loaded with or the MHCI- or MHCII-restricted OV peptides induced the secretion of IL-2 and IFN-γ, in contrast to DCs that were not loaded with antigen. Cytokine secretion was measured on day 2 (fig. 2C). OV peptide pulsed DCs inhibit tumor growth in vivo in a mouse model of lymphoma Cells from the OV-expressing mouse lymphoma cell line E.G7 were injected into C57L/6 mice on day 0 to induce tumor growth (fig. 3). fter four days the mice were immunized by injection of DCs pulsed with or the MHCI- or MHCII-restricted OV peptides, or both OV peptides in combination. s a negative control, mice were injected with untreated DCs. Six days after immunization the DCs pulsed with OV antigen already caused a slight decrease in tumor size compared to the negative control (fig. 3). Ten days after immunization there was a significant decrease in tumor OV peptide pulsed DCs induce immune respoes in a mouse model of lymphoma The tumors induced by injection of E.G7 cells were also analyzed for OV-specific cytotoxic CD8 + T cells (CTLs) with a tetramer assay (fig. 4). The numbers of CTLs were normalized to the tumor weight, which allows for the direct comparison of the individual mice. DCs pulsed with or the combination of both MHCI- and MHCIIrestricted peptides led to the highest relative numbers of OV-specific CTLs, whereas the MHCI-restricted peptide resulted in a significantly lower number of OV-specific CTLs. No OV-specific CTLs were detectable in tumors from mice treated with control DCs or DCs pulsed with the MHCII-restricted peptide (fig. 4). We also analyzed the relative titers of OVspecific IgG in the serum. The results were similar to the results from CTL enumeration, except that DCs pulsed with the MHCIIrestricted peptide also induced IgG production, just like the MHCI-restricted peptide (fig. 4). Conclusion pulsed DCs induced both CD4 + and CD8 + T cell respoes in vitro, similar to the MHCII- and MHCIrestricted peptides. Immunization of mice bearing OVexpressing tumors with DCs that were previously pulsed with or the MHCI-restricted peptide led to infiltration of OV-specific CD8 + CTLs into the tumor. In contrast, no CTLs were detectable in the tumors from mice that were treated with DCs pulsed with the MHCII-restricted peptide. This directly reflects the greater inhibition of tumor Vol 17 1/2016 MCS & more 9

5 Number of intratumor OD CTLs per mg of tumor150 (I- b ) (H-2K b ) (I- b + H-2K b ) (I- b ) (H-2K b ) (I- b + H-2K b ) Figure 4 Infiltration of CTLs into tumors and production of OV-specific IgG by DCs pulsed with OV peptides. C57L/6 mice were injected with E.G7 cells on day 0. On day 11, i.e., 7 days after immunization, single-cell suspeio prepared from tumor tissue were analyzed for OV-specific CTLs by a flow cytometric tetramer assay () and the serum was tested for OV-specific IgG by ELIS (). CTL numbers were normalized to tumor weight. n = 3; mea ± SD; Student s t-test; : non-significant at p 5; 1 p < 5; p < 1. growth by the DCs pulsed with or the MHCI-restricted peptide. pulsed DCs could induce OV-specific CD4 + T cell respoe in vivo, which resulted in the efficient production of OV-specific IgG. pulsed DCs could induce both MHCII- and MHCI-restricted T cell respoes in vivo. Therefore, antitumor effect and OV-specific CTL induction by pulsed DCs were stronger compared to MHCI- or MHCIIrestricted peptide-pulsed DCs. Respoes elicited by -pulsed DCs may be not restricted to MHCI or MHCII. MCS Product research grade Order no CD4 + T Cell Isolation Kit, mouse CD8a + T Cell Isolation Kit, mouse Mouse GM-CSF premium grade Mouse IL-4 premium grade For additional Pools, visit Order numbers are provided for 100 μg sizes. For different quality grades and additional package sizes, visit Unless otherwise specifically indicated, Miltenyi iotec products and services are for research use only and not for therapeutic or diagnostic use. 10 MCS & more Vol 17 1/2016

Supplementary Figures

Supplementary Figures Supplementary Figures Supplementary Figure 1. NKT ligand-loaded tumour antigen-presenting B cell- and monocyte-based vaccine induces NKT, NK and CD8 T cell responses. (A) The cytokine profiles of liver

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

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

Supplementary Fig. 1 p38 MAPK negatively regulates DC differentiation. (a) Western blot analysis of p38 isoform expression in BM cells, immature DCs Supplementary Fig. 1 p38 MAPK negatively regulates DC differentiation. (a) Western blot analysis of p38 isoform expression in BM cells, immature DCs (idcs) and mature DCs (mdcs). A myeloma cell line expressing

More information

Nature Medicine: doi: /nm.3922

Nature Medicine: doi: /nm.3922 Title: Glucocorticoid-induced tumor necrosis factor receptor-related protein co-stimulation facilitates tumor regression by inducing IL-9-producing helper T cells Authors: Il-Kyu Kim, Byung-Seok Kim, Choong-Hyun

More information

Therapeutic efficacy of MUC1- specific CTL and CD137 costimulation. mammary cancer model. Pinku Mukherjee & Sandra Gendler

Therapeutic efficacy of MUC1- specific CTL and CD137 costimulation. mammary cancer model. Pinku Mukherjee & Sandra Gendler Therapeutic efficacy of MUC1- specific CTL and CD137 costimulation in a spontaneous mammary cancer model Pinku Mukherjee & Sandra Gendler Goal of Immunotherapy Boosting the low level anti-tumor immune

More information

Direct ex vivo characterization of human antigen-specific CD154 + CD4 + T cells Rapid antigen-reactive T cell enrichment (Rapid ARTE)

Direct ex vivo characterization of human antigen-specific CD154 + CD4 + T cells Rapid antigen-reactive T cell enrichment (Rapid ARTE) Direct ex vivo characterization of human antigen-specific CD154 + CD4 + T cells Rapid antigen-reactive T cell enrichment (Rapid ARTE) Introduction Workflow Antigen (ag)-specific T cells play a central

More information

Dendritic cells in cancer immunotherapy Aimin Jiang

Dendritic cells in cancer immunotherapy Aimin Jiang Dendritic cells in cancer immunotherapy Aimin Jiang Feb. 11, 2014 Dendritic cells at the interface of innate and adaptive immune responses Dendritic cells: initiators of adaptive immune responses Dendritic

More information

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

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

More information

CD40L TCR IL-12 TLR-L

CD40L TCR IL-12 TLR-L CD40L B cells plasmacells Neutrophils TCR inkt cells IL-12 Ab production Can inkt cells modulate the cytokine profile of neutrophils? TLR-L CD4+ and CD8+ T cell responses 1. The invariant TCR expressed

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

Masashi Takahara,* Manami Miyai,* Mai Tomiyama,* Masato Mutou,* Andrew J. Nicol, and Mie Nieda*,1

Masashi Takahara,* Manami Miyai,* Mai Tomiyama,* Masato Mutou,* Andrew J. Nicol, and Mie Nieda*,1 Copulsing tumor antigen-pulsed dendritic cells with zoledronate efficiently enhance the expansion of tumor antigen-specific CD8 T cells via V 9 T cell activation Masashi Takahara,* Manami Miyai,* Mai Tomiyama,*

More information

W/T Itgam -/- F4/80 CD115. F4/80 hi CD115 + F4/80 + CD115 +

W/T Itgam -/- F4/80 CD115. F4/80 hi CD115 + F4/80 + CD115 + F4/8 % in the peritoneal lavage 6 4 2 p=.15 n.s p=.76 CD115 F4/8 hi CD115 + F4/8 + CD115 + F4/8 hi CD115 + F4/8 + CD115 + MHCII MHCII Supplementary Figure S1. CD11b deficiency affects the cellular responses

More information

Supplemental Figure 1

Supplemental Figure 1 Supplemental Figure 1 1a 1c PD-1 MFI fold change 6 5 4 3 2 1 IL-1α IL-2 IL-4 IL-6 IL-1 IL-12 IL-13 IL-15 IL-17 IL-18 IL-21 IL-23 IFN-α Mut Human PD-1 promoter SBE-D 5 -GTCTG- -1.2kb SBE-P -CAGAC- -1.kb

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

Supplemental Figure Legends

Supplemental Figure Legends Supplemental Figure Legends Supplemental Figure 1. SemaB / mice have normal immune cell populations. Cells were prepared from the spleens of WT and SemaB / mice, stained with various antibodies and then

More information

TITLE: MODULATION OF T CELL TOLERANCE IN A MURINE MODEL FOR IMMUNOTHERAPY OF PROSTATIC ADENOCARCINOMA

TITLE: MODULATION OF T CELL TOLERANCE IN A MURINE MODEL FOR IMMUNOTHERAPY OF PROSTATIC ADENOCARCINOMA AD Award Number: DAMD17-01-1-0085 TITLE: MODULATION OF T CELL TOLERANCE IN A MURINE MODEL FOR IMMUNOTHERAPY OF PROSTATIC ADENOCARCINOMA PRINCIPAL INVESTIGATOR: ARTHUR A HURWITZ, Ph.d. CONTRACTING ORGANIZATION:

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

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

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

Supporting Information

Supporting Information Supporting Information Bellora et al. 10.1073/pnas.1007654108 SI Materials and Methods Cells. NK cells purified from peripheral blood mononuclear cells (PBMC) of healthy donors (Human NK Cell Isolation

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION Complete but curtailed T-cell response to very-low-affinity antigen Dietmar Zehn, Sarah Y. Lee & Michael J. Bevan Supp. Fig. 1: TCR chain usage among endogenous K b /Ova reactive T cells. C57BL/6 mice

More information

Effector memory T helper cells secrete IFN-γ upon stimulation with cytokines: a role in chronic inflammation

Effector memory T helper cells secrete IFN-γ upon stimulation with cytokines: a role in chronic inflammation Excerpt from MCS&more Vol 13 1/211 Effector memory T helper cells secrete upon stimulation with cytokines: a role in chronic inflammation rne Sattler 1 *, Ulf Wagner 2, Manuela Rossol 2, Joachim Sieper

More information

Research Article Antitumor Effect of KML-B-Treated Dendritic Cells via Induction of Lymphocyte Activation

Research Article Antitumor Effect of KML-B-Treated Dendritic Cells via Induction of Lymphocyte Activation Hindawi Immunology Research Volume 217, rticle ID 2471627, 7 pages https://doi.org/1.1155/217/2471627 Research rticle ntitumor Effect of KML--Treated Dendritic Cells via Induction of Lymphocyte ctivation

More information

Tumor Immunology. Wirsma Arif Harahap Surgical Oncology Consultant

Tumor Immunology. Wirsma Arif Harahap Surgical Oncology Consultant Tumor Immunology Wirsma Arif Harahap Surgical Oncology Consultant 1) Immune responses that develop to cancer cells 2) Escape of cancer cells 3) Therapies: clinical and experimental Cancer cells can be

More information

Supplementary figure 1. Systemic delivery of anti-cd47 antibody controls tumor growth in

Supplementary figure 1. Systemic delivery of anti-cd47 antibody controls tumor growth in T u m o r v o lu m e (m m 3 ) P e rc e n t s u rv iv a l P e rc e n t s u rv iv a l Supplementary data a 1 8 6 4 2 5 1 1 5 2 2 5 3 3 5 4 T im e a fte r tu m o r in o c u la tio n (d ) b c 1 5 1 1 5 * *

More information

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

Dendritic cell subsets and CD4 T cell immunity in Melanoma. Ben Wylie 1 st year PhD Candidate Dendritic cell subsets and CD4 T cell immunity in Melanoma Ben Wylie 1 st year PhD Candidate Melanoma Melanoma is the 4 th most common cancer in Australia. Current treatment options are ineffective resulting

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi:1.138/nature1554 a TNF-α + in CD4 + cells [%] 1 GF SPF 6 b IL-1 + in CD4 + cells [%] 5 4 3 2 1 Supplementary Figure 1. Effect of microbiota on cytokine profiles of T cells in GALT. Frequencies of TNF-α

More information

Supplementary Figures

Supplementary Figures Supplementary Figures Supplementary Fig. 1. Surface thiol groups and reduction of activated T cells. (a) Activated CD8 + T-cells have high expression levels of free thiol groups on cell surface proteins.

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

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

D CD8 T cell number (x10 6 )

D CD8 T cell number (x10 6 ) IFNγ Supplemental Figure 1. CD T cell number (x1 6 ) 18 15 1 9 6 3 CD CD T cells CD6L C CD5 CD T cells CD6L D CD8 T cell number (x1 6 ) 1 8 6 E CD CD8 T cells CD6L F Log(1)CFU/g Feces 1 8 6 p

More information

Adaptive immune responses: T cell-mediated immunity

Adaptive immune responses: T cell-mediated immunity MICR2209 Adaptive immune responses: T cell-mediated immunity Dr Allison Imrie allison.imrie@uwa.edu.au 1 Synopsis: In this lecture we will discuss the T-cell mediated immune response, how it is activated,

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

Supplementary Figure 1 Maschalidi et al.

Supplementary Figure 1 Maschalidi et al. a 1% 5% % 1% 5% % OVAb βgal activity A.U. (x1 4 ) 2 1 5 βgal activity A.U. (x1 4 ) 2 1 BSAb 2 hours 4 hours OVAb BSAb OVAb BSAb,1 1 1 1 SIINFEKL (ng/ml) CFSE b Beads Alexa488 (%) 8 6 4 2 ** ** 1:1 5:1

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

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

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

Supplementary Figure 1. NAFL enhanced immunity of other vaccines (a) An over-the-counter, hand-held non-ablative fractional laser (NAFL). Supplementary Figure 1. NAFL enhanced immunity of other vaccines (a) An over-the-counter, hand-held non-ablative fractional laser (NAFL). (b) Depiction of a MTZ array generated by NAFL. (c-e) IgG production

More information

Immunotherapy for HCC

Immunotherapy for HCC Immunotherapy for HCC Jack R. Wands, MD Jeffrey and Kimberly Greenberg - Artemis and Martha Joukowsky, Professor in Gastroenterology and Professor of Medical Sciences, Director, Division of Gastroenterology

More information

08/02/59. Tumor Immunotherapy. Development of Tumor Vaccines. Types of Tumor Vaccines. Immunotherapy w/ Cytokine Gene-Transfected Tumor Cells

08/02/59. Tumor Immunotherapy. Development of Tumor Vaccines. Types of Tumor Vaccines. Immunotherapy w/ Cytokine Gene-Transfected Tumor Cells Tumor Immunotherapy Autologous virus Inactivation Inactivated virus Lymphopheresis Culture? Monocyte s Dendritic cells Immunization Autologous vaccine Development of Tumor Vaccines Types of Tumor Vaccines

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

Integrin v 3 targeted therapy for Kaposi s sarcoma with an in vitro evolved antibody 1

Integrin v 3 targeted therapy for Kaposi s sarcoma with an in vitro evolved antibody 1 Integrin v 3 targeted therapy for Kaposi s sarcoma with an in vitro evolved antibody 1 CHRISTOPH RADER, 2 MIKHAIL POPKOV, JOHN A. NEVES, AND CARLOS F. BARBAS III 2 Department of Molecular Biology and The

More information

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

L1 on PyMT tumor cells but Py117 cells are more responsive to IFN-γ. (A) Flow A MHCI B PD-L1 Fold expression 8 6 4 2 Fold expression 3 2 1 No tx 1Gy 2Gy IFN Py117 Py117 Supplementary Figure 1. Radiation and IFN-γ enhance MHCI expression and PD- L1 on PyMT tumor cells but Py117 cells

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

Supplementary Information. A vital role for IL-2 trans-presentation in DC-mediated T cell activation in humans as revealed by daclizumab therapy

Supplementary Information. A vital role for IL-2 trans-presentation in DC-mediated T cell activation in humans as revealed by daclizumab therapy Supplementary Information A vital role for IL-2 trans-presentation in DC-mediated T cell activation in humans as revealed by daclizumab therapy Simone C. Wuest 1, Jehad Edwan 1, Jayne F. Martin 1, Sungpil

More information

Understanding the T cell response to tumors using transnuclear mouse models

Understanding the T cell response to tumors using transnuclear mouse models Understanding the T cell response to tumors using transnuclear mouse models Stephanie Dougan Dana-Farber Cancer Institute Boston, MA Presenter Disclosure Information Stephanie Dougan The following relationships

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

Nature Immunology: doi: /ni Supplementary Figure 1. Gene expression profile of CD4 + T cells and CTL responses in Bcl6-deficient mice.

Nature Immunology: doi: /ni Supplementary Figure 1. Gene expression profile of CD4 + T cells and CTL responses in Bcl6-deficient mice. Supplementary Figure 1 Gene expression profile of CD4 + T cells and CTL responses in Bcl6-deficient mice. (a) Gene expression profile in the resting CD4 + T cells were analyzed by an Affymetrix microarray

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

PRIME-XV Dendritic Cell Maturation CDM

PRIME-XV Dendritic Cell Maturation CDM PRIME-XV Dendritic Cell Maturation CDM Chemically defined, animal component-free medium for dendritic cell culture Optimized for differentiation of monocytes into immature dendritic cells (idcs) and subsequent

More information

Madhav V. Dhodapkar, Joseph Krasovsky, Ralph M. Steinman, and Nina Bhardwaj

Madhav V. Dhodapkar, Joseph Krasovsky, Ralph M. Steinman, and Nina Bhardwaj Mature dendritic cells boost functionally superior CD8 + T-cell in humans without foreign helper epitopes Rapid PUBLICATION Madhav V. Dhodapkar, Joseph Krasovsky, Ralph M. Steinman, and Nina Bhardwaj Laboratory

More information

TITLE: Modulation of T Cell Tolerance in a Murine Model for Immunotheraphy of Prostatic Adenocarcinoma

TITLE: Modulation of T Cell Tolerance in a Murine Model for Immunotheraphy of Prostatic Adenocarcinoma AD Award Number: DAMD17-01-1-0085 TITLE: Modulation of T Cell Tolerance in a Murine Model for Immunotheraphy of Prostatic Adenocarcinoma PRINCIPAL INVESTIGATOR: Arthur A. Hurwitz, Ph.D. CONTRACTING ORGANIZATION:

More information

In vitro human regulatory T cell suppression assay

In vitro human regulatory T cell suppression assay Human CD4 + CD25 + regulatory T cell isolation, in vitro suppression assay and analysis In vitro human regulatory T cell suppression assay Introduction Regulatory T (Treg) cells are a subpopulation of

More information

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

Supplementary Figure 1. Deletion of Smad3 prevents B16F10 melanoma invasion and metastasis in a mouse s.c. tumor model. A B16F1 s.c. Lung LN Distant lymph nodes Colon B B16F1 s.c. Supplementary Figure 1. Deletion of Smad3 prevents B16F1 melanoma invasion and metastasis in a mouse s.c. tumor model. Highly invasive growth

More information

PREPARED FOR: U.S. Army Medical Research and Materiel Command Fort Detrick, Maryland Approved for public release; distribution unlimited

PREPARED FOR: U.S. Army Medical Research and Materiel Command Fort Detrick, Maryland Approved for public release; distribution unlimited AD (Leave blank) Award Number: W81XWH-07-1-0345 TITLE: Second-Generation Therapeutic DNA Lymphoma Vaccines PRINCIPAL INVESTIGATOR: Larry W. Kwak, M.D., Ph.D. CONTRACTING ORGANIZATION: University of Texas

More information

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

ACTIVATION AND EFFECTOR FUNCTIONS OF CELL-MEDIATED IMMUNITY AND NK CELLS. Choompone Sakonwasun, MD (Hons), FRCPT ACTIVATION AND EFFECTOR FUNCTIONS OF CELL-MEDIATED IMMUNITY AND NK CELLS Choompone Sakonwasun, MD (Hons), FRCPT Types of Adaptive Immunity Types of T Cell-mediated Immune Reactions CTLs = cytotoxic T lymphocytes

More information

Supporting Information

Supporting Information Supporting Information Shime et al. 1.173/pnas.11139919 SI Methods Reagents. was purchased from GE Healthcare, which was free from LPS contamination. TNF-α and IFN-β ELISA kit was purchased from eioscience

More information

Supporting Information

Supporting Information Supporting Information lpek et al. 1.173/pnas.1121217 SI Materials and Methods Mice. cell knockout, inos / (Taconic arms), Rag1 /, INγR /, and IL-12p4 / mice (The Jackson Laboratory) were maintained and/or

More information

Supplementary Materials for

Supplementary Materials for www.sciencetranslationalmedicine.org/cgi/content/full/8/352/352ra110/dc1 Supplementary Materials for Spatially selective depletion of tumor-associated regulatory T cells with near-infrared photoimmunotherapy

More information

Regulation of anti-tumor immunity through migration of immune cell subsets within the tumor microenvironment Thomas F. Gajewski, M.D., Ph.D.

Regulation of anti-tumor immunity through migration of immune cell subsets within the tumor microenvironment Thomas F. Gajewski, M.D., Ph.D. Regulation of anti-tumor immunity through migration of immune cell subsets within the tumor microenvironment Thomas F. Gajewski, M.D., Ph.D. Professor, Departments of Pathology and Medicine Program Leader,

More information

Tumors arise from accumulated genetic mutations. Tumor Immunology (Cancer)

Tumors arise from accumulated genetic mutations. Tumor Immunology (Cancer) Tumor Immunology (Cancer) Tumors arise from accumulated genetic mutations Robert Beatty MCB150 Mutations Usually have >6 mutations in both activation/growth factors and tumor suppressor genes. Types of

More information

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

IOM Immunization Safety Review 11/12/2001. Immunological Competition and the Infant Immune Response to Vaccines IOM Immunization Safety Review 11/12/2001 Immunological Competition and the Infant Immune Response to Vaccines Richard Insel University of Rochester Goals Neonatal and Infant Immune System Broad Effects

More information

Supplementary Figure 1.

Supplementary Figure 1. Supplementary Figure 1. Female Pro-ins2 -/- mice at 5-6 weeks of age were either inoculated i.p. with a single dose of CVB4 (1x10 5 PFU/mouse) or PBS and treated with αgalcer or control vehicle. On day

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

Using Multiplex Assays and Assay Development to Enhance your Research Program James A. Lederer, PhD

Using Multiplex Assays and Assay Development to Enhance your Research Program James A. Lederer, PhD Using Multiplex Assays and Assay Development to Enhance your Research Program James A. Lederer, PhD Department of Surgery Brigham and Women s Hospital (BWH), BWH Biomedical Research Institute (BWH-BRI)

More information

Immunology. T-Lymphocytes. 16. Oktober 2014, Ruhr-Universität Bochum Karin Peters,

Immunology. T-Lymphocytes. 16. Oktober 2014, Ruhr-Universität Bochum Karin Peters, Immunology T-Lymphocytes 16. Oktober 2014, Ruhr-Universität Bochum Karin Peters, karin.peters@rub.de The role of T-effector cells in the immune response against microbes cellular immunity humoral immunity

More information

Radiation Therapy as an Immunomodulator

Radiation Therapy as an Immunomodulator Radiation Therapy as an Immunomodulator Yvonne Mowery, MD, PhD February 20, 2017 Tumor/Immune System Balance Kalbasi, JCI 2013 UNC-Duke-NC State-Wake Forest Spring 2017 2 RT Can Shift Balance Toward Elimination

More information

Examples of questions for Cellular Immunology/Cellular Biology and Immunology

Examples of questions for Cellular Immunology/Cellular Biology and Immunology Examples of questions for Cellular Immunology/Cellular Biology and Immunology Each student gets a set of 6 questions, so that each set contains different types of questions and that the set of questions

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

Mechanismen der allergenspezifischen Immuntherapie

Mechanismen der allergenspezifischen Immuntherapie Mechanismen der allergenspezifischen Immuntherapie Medizinische Universität Wien Zentrum für Pathophysiologie, Infektiologie und Immunologie Institut für Pathophysiologie und Allergieforschung Abteilung

More information

Supplementary Materials for

Supplementary Materials for www.sciencetranslationalmedicine.org/cgi/content/full/8/333/333ra47/dc1 Supplementary Materials for Androgen receptor antagonists compromise T cell response against prostate cancer leading to early tumor

More 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

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 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 15 min at 37 C and replaced with fresh complete medium.

More information

Automated manufacture of T-cell immunotherapies using gamma retroviral transduction. Lee Markwick, PhD. AMC, Manchester March 2018

Automated manufacture of T-cell immunotherapies using gamma retroviral transduction. Lee Markwick, PhD. AMC, Manchester March 2018 Automated manufacture of T-cell immunotherapies using gamma retroviral transduction Lee Markwick, PhD AMC, Manchester March 2018 Automated generation of CAR T cells Starting material d0 T cell enrichment

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 Programmed death one homolog maintains the pool size of regulatory T cells by promoting their differentiation and stability Qi Wang 1, Jianwei He 1, Dallas B. Flies 2, Liqun Luo

More information

Supporting Information Table of Contents

Supporting Information Table of Contents Supporting Information Table of Contents Supporting Information Figure 1 Page 2 Supporting Information Figure 2 Page 4 Supporting Information Figure 3 Page 5 Supporting Information Figure 4 Page 6 Supporting

More information

Generation of monocytederived Dendritic Cells (modcs)

Generation of monocytederived Dendritic Cells (modcs) monocytederived Dendritic (modcs) Application Note Background Dendritic (DCs) are so called because of their characteristic cell surface projections that resemble the dendrites of neurons (see Fig 1 and

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

Supplemental Figure 1. Protein L

Supplemental Figure 1. Protein L Supplemental Figure 1 Protein L m19delta T m1928z T Suppl. Fig 1. Expression of CAR: B6-derived T cells were transduced with m19delta (left) and m1928z (right) to generate CAR T cells and transduction

More information

Defensive mechanisms include :

Defensive mechanisms include : Acquired Immunity Defensive mechanisms include : 1) Innate immunity (Natural or Non specific) 2) Acquired immunity (Adaptive or Specific) Cell-mediated immunity Humoral immunity Two mechanisms 1) Humoral

More information

TITLE: Development of Antigen Presenting Cells for adoptive immunotherapy in prostate cancer

TITLE: Development of Antigen Presenting Cells for adoptive immunotherapy in prostate cancer AD Award Number: W8-XWH-5-- TITLE: Development of Antigen Presenting Cells for adoptive immunotherapy in prostate cancer PRINCIPAL INVESTIGATOR: Mathias Oelke, Ph.D. CONTRACTING ORGANIZATION: Johns Hopkins

More information

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

Combined Rho-kinase inhibition and immunogenic cell death triggers and propagates immunity against cancer Supplementary Information Combined Rho-kinase inhibition and immunogenic cell death triggers and propagates immunity against cancer Gi-Hoon Nam, Eun-Jung Lee, Yoon Kyoung Kim, Yeonsun Hong, Yoonjeong Choi,

More information

Dendritic Cell Based Immunotherapy for Cancer. Edgar G. Engleman, M.D.

Dendritic Cell Based Immunotherapy for Cancer. Edgar G. Engleman, M.D. Dendritic Cell Based Immunotherapy for Cancer Edgar G. Engleman, M.D. Two main DC subsets Myeloid (mydc) Derived from monocytes Capture/process/present Ag to T cells Activate NK cells and B cells Plasmacytoid

More information

Vasoactive intestinal peptide inhibits IFN-α secretion and modulates the immune function of plasmacytoid dendritic cells

Vasoactive intestinal peptide inhibits IFN-α secretion and modulates the immune function of plasmacytoid dendritic cells Excerpt from MACS&more Vol 12 1/21 Vasoactive intestinal peptide inhibits IFN-α secretion and modulates the immune function of plasmacytoid dendritic cells Dorit Fabricius 1, 2,, M. Sue O Dorisio 1, 2,

More information

C. Incorrect! MHC class I molecules are not involved in the process of bridging in ADCC.

C. Incorrect! MHC class I molecules are not involved in the process of bridging in ADCC. Immunology - Problem Drill 13: T- Cell Mediated Immunity Question No. 1 of 10 1. During Antibody-dependent cell mediated cytotoxicity (ADCC), the antibody acts like a bridge between the specific antigen

More information

Conventional anti-cancer therapies (surgery and radio- and

Conventional anti-cancer therapies (surgery and radio- and The Immunogenicity of Dendritic Cell-Based Vaccines Is Not Hampered by Doxorubicin and Melphalan Administration 1 Anna Casati, 2 Valérie S. Zimmermann, 2 Fabio Benigni, 3 Maria T. S. Bertilaccio, Matteo

More information

Rapid antigen-specific T cell enrichment (Rapid ARTE)

Rapid antigen-specific T cell enrichment (Rapid ARTE) Direct ex vivo characterization of human antigen-specific CD154+CD4+ T cell Rapid antigen-specific T cell enrichment (Rapid ARTE) Introduction Workflow Antigen (ag)-specific T cells play a central role

More information

Gp96-Ig-SIV VACCINES INDUCE PREDOMINANT IMMUNE RESPONSES AT MUCOSAL SITES

Gp96-Ig-SIV VACCINES INDUCE PREDOMINANT IMMUNE RESPONSES AT MUCOSAL SITES Gp96-Ig-SIV VACCINES INDUCE PREDOMINANT IMMUNE RESPONSES AT MUCOSAL SITES Natasa Strbo M.D. Ph.D. Paris, AIDS Vaccine 29 October 19 22 Why is cell secreted gp96 Ig vaccine approach innovative and unconventional?

More information

Regulation of Type 2 Immunity by Basophils Prof. Dr. David Voehringer

Regulation of Type 2 Immunity by Basophils Prof. Dr. David Voehringer Regulation of Type 2 Immunity by Basophils Department of Infection Biology Institute of Clinical Microbiology, Immunology and Hygiene Outline of the presentation The concept of type 2 immunity Basophil

More information

They determine if there will be an immune response. Determine functions associated with immune response, but not specific to Ag.

They determine if there will be an immune response. Determine functions associated with immune response, but not specific to Ag. Appendices A They determine if there will be an immune response. Antigen receptor genes in T cells (TCR) and B cell (Ig) Determine functions associated with immune response, but not specific to Ag. MHC

More information

Vaccination with OVA-bound nanoparticles encapsulating IL-7 inhibits the growth of OVA-expressing E.G7 tumor cells in vivo

Vaccination with OVA-bound nanoparticles encapsulating IL-7 inhibits the growth of OVA-expressing E.G7 tumor cells in vivo 292 Vaccination with OVA-bound nanoparticles encapsulating IL-7 inhibits the growth of OVA-expressing E.G7 tumor cells in vivo HIROKO TOYOTA 1, NORIKO YANASE 1, TAKAYUKI YOSHIMOTO 2, MITSUNORI HARADA 3,

More information

MACS Marker Screen. MACS Marker Screen. Catalog antibodies. antibodies CD4 (VIT4) CD11b. MACS Marker Screen antibodies. Catalog. antibodies CD2 CD14

MACS Marker Screen. MACS Marker Screen. Catalog antibodies. antibodies CD4 (VIT4) CD11b. MACS Marker Screen antibodies. Catalog. antibodies CD2 CD14 MACS Marker Screen The performance of MACS Marker Screen was tested and compared with catalog reagents offered in single vials as liquid antibody reagents. The were reconstituted using 25 µl of deionized

More information

Zheng, BJ; Du, LY; Zhao, GY; Lin, YP; Sui, HY; Chan, C; Ma, S; Guan, Y; Yuen, KY. Citation Hong Kong Medical Journal, 2008, v. 14 suppl. 4, p.

Zheng, BJ; Du, LY; Zhao, GY; Lin, YP; Sui, HY; Chan, C; Ma, S; Guan, Y; Yuen, KY. Citation Hong Kong Medical Journal, 2008, v. 14 suppl. 4, p. Title Studies of SARS virus vaccines Author(s) Zheng, BJ; Du, LY; Zhao, GY; Lin, YP; Sui, HY; Chan, C; Ma, S; Guan, Y; Yuen, KY Citation Hong Kong Medical Journal, 2008, v. 14 suppl. 4, p. 39-43 Issued

More information

Figure S1. Western blot analysis of clathrin RNA interference in human DCs Human immature DCs were transfected with 100 nm Clathrin SMARTpool or

Figure S1. Western blot analysis of clathrin RNA interference in human DCs Human immature DCs were transfected with 100 nm Clathrin SMARTpool or Figure S1. Western blot analysis of clathrin RNA interference in human DCs Human immature DCs were transfected with 100 nm Clathrin SMARTpool or control nontargeting sirnas. At 90 hr after transfection,

More information

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

Hua Tang, Weiping Cao, Sudhir Pai Kasturi, Rajesh Ravindran, Helder I Nakaya, Kousik SUPPLEMENTARY FIGURES 1-19 T H 2 response to cysteine-proteases requires dendritic cell-basophil cooperation via ROS mediated signaling Hua Tang, Weiping Cao, Sudhir Pai Kasturi, Rajesh Ravindran, Helder

More information

Rationale for Combining Immunotherapy with Chemotherapy or Targeted Therapy

Rationale for Combining Immunotherapy with Chemotherapy or Targeted Therapy Society for Immunotherapy of Cancer (SITC) Rationale for Combining Immunotherapy with Chemotherapy or Targeted Therapy Qing Yi, MD, PhD Staff and Chair, Department of Cancer Biology Betsy B. DeWindt Endowed

More information

SUPPLEMENTARY INFORMATION

SUPPLEMENTARY INFORMATION doi: 1.138/nature775 4 O.D. (595-655) 3 1 -ζ no antibody isotype ctrl Plated Soluble 1F6 397 7H11 Supplementary Figure 1 Soluble and plated anti- Abs induce -! signalling. B3Z cells stably expressing!

More information

Immune Checkpoints. PD Dr med. Alessandra Curioni-Fontecedro Department of Hematology and Oncology Cancer Center Zurich University Hospital Zurich

Immune Checkpoints. PD Dr med. Alessandra Curioni-Fontecedro Department of Hematology and Oncology Cancer Center Zurich University Hospital Zurich Immune Checkpoints PD Dr med. Alessandra Curioni-Fontecedro Department of Hematology and Oncology Cancer Center Zurich University Hospital Zurich Activation of T cells requires co-stimulation Science 3

More information

Immunological Monitoring of Cancer Vaccine Clinical Trials:

Immunological Monitoring of Cancer Vaccine Clinical Trials: Immunological Monitoring of Cancer Vaccine Clinical Trials: Why The what Lab, do to we measure, the need Tasks a how centralized and and the why? Tests lab? Anatoli Malyguine, M.D., Ph.D. Laboratory of

More information

Epitope discovery. Using predicted MHC binding CENTER FOR BIOLOGICAL SEQUENCE ANALYSIS

Epitope discovery. Using predicted MHC binding CENTER FOR BIOLOGICAL SEQUENCE ANALYSIS Epitope discovery Using predicted MHC binding SARS corona virus The 2003 outbreak The disease Lung Pathology Inflammatory exudation in alveoli and interstitial spaces Monocytic and lymphocytic infiltration

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