Antibody data
- Antibody Data
- Antigen structure
- References [61]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [35]
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- Product number
- 45-5825-82 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- T-bet Monoclonal Antibody (eBio4B10 (4B10)), PerCP-Cyanine5.5, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The eBio4B10 monoclonal antibody reacts with mouse and human T-bet. T-bet is a Th1-specific T-box transcription factor critical to the development of the Th1 CD4+ T cell lineage. This is known based on the observations that T-Bet deficient mice have impaired Th1 cell development, and that ectopic expression of T-Bet results in development skewed to the Th1 lineage. T-Bet expression is induced by the Th1 cytokine IFN gamma, and T-Bet also regulates the expression of IFN gamma, likely, at least in part, through the modification of DNA accessibility and histone remodeling. In addition to IFN gamma, T-Bet is also known to regulate the expression of IL-12R beta and IL-2. Moreover, T-Bet plays a role in class-switch recombination in B-cells. Applications Reported: This eBio4B10 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This eBio4B10 antibody has been tested by intracellular staining and flow cytometric analysis of normal human peripheral blood cells. This antibody can be used at less than or equal to 0.25 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. For intracellular staining with the eBio4B10 monoclonal antibody, it is recommended to use the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to Best Protocols; Protocol B: One step protocol for intracellular (nuclear) proteins). Excitation: 488 nm; Emission: 695 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- eBio4B10 (4B10)
- Vial size
- 100 µg
- Concentration
- 0.2 mg/mL
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Graded RhoA GTPase Expression in Treg Cells Distinguishes Tumor Immunity From Autoimmunity.
An AAV-based, room-temperature-stable, single-dose COVID-19 vaccine provides durable immunogenicity and protection in non-human primates.
Heterologous arenavirus vector prime-boost overrules self-tolerance for efficient tumor-specific CD8 T cell attack.
Culture, expansion, and flow-cytometry-based functional analysis of pteropid bat MR1-restricted unconventional T cells.
Francisella tularensis induces Th1 like MAIT cells conferring protection against systemic and local infection.
Immunogenicity of an AAV-based, room-temperature stable, single dose COVID-19 vaccine in mice and non-human primates.
Targeting Phosphatidylserine Enhances the Anti-tumor Response to Tumor-Directed Radiation Therapy in a Preclinical Model of Melanoma.
ADAM12 is a costimulatory molecule that determines Th1 cell fate and mediates tissue inflammation.
Imbalance between T helper 1 and regulatory T cells plays a detrimental role in experimental Parkinson's disease in mice.
Th2 Biased Immunity With Altered B Cell Profiles in Circulation of Patients With Sporotrichosis Caused by Sporothrix globosa.
MiR-1165-3p Suppresses Th2 Differentiation via Targeting IL-13 and PPM1A in a Mouse Model of Allergic Airway Inflammation.
Programming Multifaceted Pulmonary T Cell Immunity by Combination Adjuvants.
CD19-CAR-T Cells Bearing a KIR/PD-1-Based Inhibitory CAR Eradicate CD19(+)HLA-C1(-) Malignant B Cells While Sparing CD19(+)HLA-C1(+) Healthy B Cells.
Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity.
Adjuvant-free nanofiber vaccine induces in situ lung dendritic cell activation and T(H)17 responses.
The Obese Liver Environment Mediates Conversion of NK Cells to a Less Cytotoxic ILC1-Like Phenotype.
Characterization of Transcriptional Regulatory Networks that Promote and Restrict Identities and Functions of Intestinal Innate Lymphoid Cells.
Single-Cell Profiling Defines Transcriptomic Signatures Specific to Tumor-Reactive versus Virus-Responsive CD4(+) T Cells.
FCRL5(+) Memory B Cells Exhibit Robust Recall Responses.
Keap1-Nrf2 System Plays an Important Role in Invariant Natural Killer T Cell Development and Homeostasis.
Limited Foxp3(+) Regulatory T Cells Response During Acute Trypanosoma cruzi Infection Is Required to Allow the Emergence of Robust Parasite-Specific CD8(+) T Cell Immunity.
Effects of Anti-Integrin Treatment With Vedolizumab on Immune Pathways and Cytokines in Inflammatory Bowel Diseases.
TRPM2 Exacerbates Central Nervous System Inflammation in Experimental Autoimmune Encephalomyelitis by Increasing Production of CXCL2 Chemokines.
CD38-NAD(+)Axis Regulates Immunotherapeutic Anti-Tumor T Cell Response.
Initiation of Antiviral B Cell Immunity Relies on Innate Signals from Spatially Positioned NKT Cells.
Regulation of chitinase-3-like-1 in T cell elicits Th1 and cytotoxic responses to inhibit lung metastasis.
Conventional NK cells and ILC1 are partially ablated in the livers of Ncr1 (iCre)Tbx21 (fl/fl) mice.
Antibiotic-induced perturbations in microbial diversity during post-natal development alters amyloid pathology in an aged APP(SWE)/PS1(ΔE9) murine model of Alzheimer's disease.
Resistance to TGFβ suppression and improved anti-tumor responses in CD8(+) T cells lacking PTPN22.
The NLRP3 inflammasome contributes to host protection during Sporothrix schenckii infection.
CD36 receptor regulates malaria-induced immune responses primarily at early blood stage infection contributing to parasitemia control and resistance to mortality.
B-cell anergy induces a Th17 shift in a novel B lymphocyte transgenic NOD mouse model, the 116C-NOD mouse.
CXCL16-positive dendritic cells enhance invariant natural killer T cell-dependent IFNγ production and tumor control.
IL12-mediated sensitizing of T-cell receptor-dependent and -independent tumor cell killing.
Human Head and Neck Squamous Cell Carcinoma-Associated Semaphorin 4D Induces Expansion of Myeloid-Derived Suppressor Cells.
Oxygen Sensing by T Cells Establishes an Immunologically Tolerant Metastatic Niche.
Intranasal Administration of Lentiviral miR-135a Regulates Mast Cell and Allergen-Induced Inflammation by Targeting GATA-3.
Chronic Trichuris muris Infection Decreases Diversity of the Intestinal Microbiota and Concomitantly Increases the Abundance of Lactobacilli.
Eliminating roles for T-bet and IL-2 but revealing superior activation and proliferation as mechanisms underpinning dominance of regulatory T cells in tumors.
Eliminating roles for T-bet and IL-2 but revealing superior activation and proliferation as mechanisms underpinning dominance of regulatory T cells in tumors.
IL-10 inhibits neuraminidase-activated TGF-β and facilitates Th1 phenotype during early phase of infection.
Expression of GM-CSF in T Cells Is Increased in Multiple Sclerosis and Suppressed by IFN-β Therapy.
The MAPK-Activated Kinase MK2 Attenuates Dendritic Cell-Mediated Th1 Differentiation and Autoimmune Encephalomyelitis.
The transduction pattern of IL-12-encoding lentiviral vectors shapes the immunological outcome.
T cell factor-1 controls the lifetime of CD4+ CD8+ thymocytes in vivo and distal T cell receptor α-chain rearrangement required for NKT cell development.
Prophylaxis with a respiratory syncytial virus (RSV) anti-G protein monoclonal antibody shifts the adaptive immune response to RSV rA2-line19F infection from Th2 to Th1 in BALB/c mice.
High eomesodermin expression among CD57+ CD8+ T cells identifies a CD8+ T cell subset associated with viral control during chronic human immunodeficiency virus infection.
MicroRNA-155 is essential for T(H)2-mediated allergen-induced eosinophilic inflammation in the lung.
CD4CD8αα lymphocytes, a novel human regulatory T cell subset induced by colonic bacteria and deficient in patients with inflammatory bowel disease.
Alterations in regulatory T cells induced by specific oligosaccharides improve vaccine responsiveness in mice.
Proinflammatory Vδ2+ T cells populate the human intestinal mucosa and enhance IFN-γ production by colonic αβ T cells.
Helicobacter pylori infection in a pig model is dominated by Th1 and cytotoxic CD8+ T cell responses.
Systemic 4-1BB activation induces a novel T cell phenotype driven by high expression of Eomesodermin.
Breakdown in repression of IFN-γ mRNA leads to accumulation of self-reactive effector CD8+ T cells.
Increase in Th17 and T-reg lymphocytes and decrease of IL22 correlate with the recovery phase of acute EAE in rat.
Differential requirement for the CD45 splicing regulator hnRNPLL for accumulation of NKT and conventional T cells.
DOCK8 deficiency impairs CD8 T cell survival and function in humans and mice.
CD134 plus CD137 dual costimulation induces Eomesodermin in CD4 T cells to program cytotoxic Th1 differentiation.
Cytokine requirements for the differentiation and expansion of IL-17A- and IL-22-producing human Vgamma2Vdelta2 T cells.
Differential expression of T-bet, a T-box transcription factor required for Th1 T-cell development, in peripheral T-cell lymphomas.
A novel transcription factor, T-bet, directs Th1 lineage commitment.
Kalim KW, Yang JQ, Modur V, Nguyen P, Li Y, Zheng Y, Guo F
Frontiers in immunology 2021;12:726393
Frontiers in immunology 2021;12:726393
An AAV-based, room-temperature-stable, single-dose COVID-19 vaccine provides durable immunogenicity and protection in non-human primates.
Zabaleta N, Dai W, Bhatt U, Hérate C, Maisonnasse P, Chichester JA, Sanmiguel J, Estelien R, Michalson KT, Diop C, Maciorowski D, Dereuddre-Bosquet N, Cavarelli M, Gallouët AS, Naninck T, Kahlaoui N, Lemaitre J, Qi W, Hudspeth E, Cucalon A, Dyer CD, Pampena MB, Knox JJ, LaRocque RC, Charles RC, Li D, Kim M, Sheridan A, Storm N, Johnson RI, Feldman J, Hauser BM, Contreras V, Marlin R, Tsong Fang RH, Chapon C, van der Werf S, Zinn E, Ryan A, Kobayashi DT, Chauhan R, McGlynn M, Ryan ET, Schmidt AG, Price B, Honko A, Griffiths A, Yaghmour S, Hodge R, Betts MR, Freeman MW, Wilson JM, Le Grand R, Vandenberghe LH
Cell host & microbe 2021 Sep 8;29(9):1437-1453.e8
Cell host & microbe 2021 Sep 8;29(9):1437-1453.e8
Heterologous arenavirus vector prime-boost overrules self-tolerance for efficient tumor-specific CD8 T cell attack.
Bonilla WV, Kirchhammer N, Marx AF, Kallert SM, Krzyzaniak MA, Lu M, Darbre S, Schmidt S, Raguz J, Berka U, Vincenti I, Pauzuolis M, Kerber R, Hoepner S, Günther S, Magnus C, Merkler D, Orlinger KK, Zippelius A, Pinschewer DD
Cell reports. Medicine 2021 Mar 16;2(3):100209
Cell reports. Medicine 2021 Mar 16;2(3):100209
Culture, expansion, and flow-cytometry-based functional analysis of pteropid bat MR1-restricted unconventional T cells.
Sia WR, Hey YY, Foo R, Wang LF, Leeansyah E
STAR protocols 2021 Jun 18;2(2):100487
STAR protocols 2021 Jun 18;2(2):100487
Francisella tularensis induces Th1 like MAIT cells conferring protection against systemic and local infection.
Zhao Z, Wang H, Shi M, Zhu T, Pediongco T, Lim XY, Meehan BS, Nelson AG, Fairlie DP, Mak JYW, Eckle SBG, de Lima Moreira M, Tumpach C, Bramhall M, Williams CG, Lee HJ, Haque A, Evrard M, Rossjohn J, McCluskey J, Corbett AJ, Chen Z
Nature communications 2021 Jul 16;12(1):4355
Nature communications 2021 Jul 16;12(1):4355
Immunogenicity of an AAV-based, room-temperature stable, single dose COVID-19 vaccine in mice and non-human primates.
Zabaleta N, Dai W, Bhatt U, Chichester JA, Sanmiguel J, Estelien R, Michalson KT, Diop C, Maciorowski D, Qi W, Hudspeth E, Cucalon A, Dyer CD, Pampena MB, Knox JJ, LaRocque RC, Charles RC, Li D, Kim M, Sheridan A, Storm N, Johnson RI, Feldman J, Hauser BM, Zinn E, Ryan A, Kobayashi DT, Chauhan R, McGlynn M, Ryan ET, Schmidt AG, Price B, Honko A, Griffiths A, Yaghmour S, Hodge R, Betts MR, Freeman MW, Wilson JM, Vandenberghe LH
bioRxiv : the preprint server for biology 2021 Jan 19;
bioRxiv : the preprint server for biology 2021 Jan 19;
Targeting Phosphatidylserine Enhances the Anti-tumor Response to Tumor-Directed Radiation Therapy in a Preclinical Model of Melanoma.
Budhu S, Giese R, Gupta A, Fitzgerald K, Zappasodi R, Schad S, Hirschhorn D, Campesato LF, De Henau O, Gigoux M, Liu C, Mazo G, Deng L, Barker CA, Wolchok JD, Merghoub T
Cell reports 2021 Jan 12;34(2):108620
Cell reports 2021 Jan 12;34(2):108620
ADAM12 is a costimulatory molecule that determines Th1 cell fate and mediates tissue inflammation.
Liu Y, Bockermann R, Hadi M, Safari I, Carrion B, Kveiborg M, Issazadeh-Navikas S
Cellular & molecular immunology 2021 Aug;18(8):1904-1919
Cellular & molecular immunology 2021 Aug;18(8):1904-1919
Imbalance between T helper 1 and regulatory T cells plays a detrimental role in experimental Parkinson's disease in mice.
Li W, Luo Y, Xu H, Ma Q, Yao Q
The Journal of international medical research 2021 Apr;49(4):300060521998471
The Journal of international medical research 2021 Apr;49(4):300060521998471
Th2 Biased Immunity With Altered B Cell Profiles in Circulation of Patients With Sporotrichosis Caused by Sporothrix globosa.
Zu J, Yao L, Song Y, Cui Y, Guan M, Chen R, Zhen Y, Li S
Frontiers in immunology 2020;11:570888
Frontiers in immunology 2020;11:570888
MiR-1165-3p Suppresses Th2 Differentiation via Targeting IL-13 and PPM1A in a Mouse Model of Allergic Airway Inflammation.
Wang Z, Ji N, Chen Z, Sun Z, Wu C, Yu W, Hu F, Huang M, Zhang M
Allergy, asthma & immunology research 2020 Sep;12(5):859-876
Allergy, asthma & immunology research 2020 Sep;12(5):859-876
Programming Multifaceted Pulmonary T Cell Immunity by Combination Adjuvants.
Marinaik CB, Kingstad-Bakke B, Lee W, Hatta M, Sonsalla M, Larsen A, Neldner B, Gasper DJ, Kedl RM, Kawaoka Y, Suresh M
Cell reports. Medicine 2020 Sep 22;1(6):100095
Cell reports. Medicine 2020 Sep 22;1(6):100095
CD19-CAR-T Cells Bearing a KIR/PD-1-Based Inhibitory CAR Eradicate CD19(+)HLA-C1(-) Malignant B Cells While Sparing CD19(+)HLA-C1(+) Healthy B Cells.
Tao L, Farooq MA, Gao Y, Zhang L, Niu C, Ajmal I, Zhou Y, He C, Zhao G, Yao J, Liu M, Jiang W
Cancers 2020 Sep 13;12(9)
Cancers 2020 Sep 13;12(9)
Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity.
Kimura S, Nakamura Y, Kobayashi N, Shiroguchi K, Kawakami E, Mutoh M, Takahashi-Iwanaga H, Yamada T, Hisamoto M, Nakamura M, Udagawa N, Sato S, Kaisho T, Iwanaga T, Hase K
Nature communications 2020 Jan 13;11(1):234
Nature communications 2020 Jan 13;11(1):234
Adjuvant-free nanofiber vaccine induces in situ lung dendritic cell activation and T(H)17 responses.
Si Y, Tian Q, Zhao F, Kelly SH, Shores LS, Camacho DF, Sperling AI, Andrade MS, Collier JH, Chong AS
Science advances 2020 Aug;6(32):eaba0995
Science advances 2020 Aug;6(32):eaba0995
The Obese Liver Environment Mediates Conversion of NK Cells to a Less Cytotoxic ILC1-Like Phenotype.
Cuff AO, Sillito F, Dertschnig S, Hall A, Luong TV, Chakraverty R, Male V
Frontiers in immunology 2019;10:2180
Frontiers in immunology 2019;10:2180
Characterization of Transcriptional Regulatory Networks that Promote and Restrict Identities and Functions of Intestinal Innate Lymphoid Cells.
Pokrovskii M, Hall JA, Ochayon DE, Yi R, Chaimowitz NS, Seelamneni H, Carriero N, Watters A, Waggoner SN, Littman DR, Bonneau R, Miraldi ER
Immunity 2019 Jul 16;51(1):185-197.e6
Immunity 2019 Jul 16;51(1):185-197.e6
Single-Cell Profiling Defines Transcriptomic Signatures Specific to Tumor-Reactive versus Virus-Responsive CD4(+) T Cells.
Magen A, Nie J, Ciucci T, Tamoutounour S, Zhao Y, Mehta M, Tran B, McGavern DB, Hannenhalli S, Bosselut R
Cell reports 2019 Dec 3;29(10):3019-3032.e6
Cell reports 2019 Dec 3;29(10):3019-3032.e6
FCRL5(+) Memory B Cells Exhibit Robust Recall Responses.
Kim CC, Baccarella AM, Bayat A, Pepper M, Fontana MF
Cell reports 2019 Apr 30;27(5):1446-1460.e4
Cell reports 2019 Apr 30;27(5):1446-1460.e4
Keap1-Nrf2 System Plays an Important Role in Invariant Natural Killer T Cell Development and Homeostasis.
Pyaram K, Kumar A, Kim YH, Noel S, Reddy SP, Rabb H, Chang CH
Cell reports 2019 Apr 16;27(3):699-707.e4
Cell reports 2019 Apr 16;27(3):699-707.e4
Limited Foxp3(+) Regulatory T Cells Response During Acute Trypanosoma cruzi Infection Is Required to Allow the Emergence of Robust Parasite-Specific CD8(+) T Cell Immunity.
Araujo Furlan CL, Tosello Boari J, Rodriguez C, Canale FP, Fiocca Vernengo F, Boccardo S, Beccaria CG, Adoue V, Joffre O, Gruppi A, Montes CL, Acosta Rodriguez EV
Frontiers in immunology 2018;9:2555
Frontiers in immunology 2018;9:2555
Effects of Anti-Integrin Treatment With Vedolizumab on Immune Pathways and Cytokines in Inflammatory Bowel Diseases.
Rath T, Billmeier U, Ferrazzi F, Vieth M, Ekici A, Neurath MF, Atreya R
Frontiers in immunology 2018;9:1700
Frontiers in immunology 2018;9:1700
TRPM2 Exacerbates Central Nervous System Inflammation in Experimental Autoimmune Encephalomyelitis by Increasing Production of CXCL2 Chemokines.
Tsutsui M, Hirase R, Miyamura S, Nagayasu K, Nakagawa T, Mori Y, Shirakawa H, Kaneko S
The Journal of neuroscience : the official journal of the Society for Neuroscience 2018 Sep 26;38(39):8484-8495
The Journal of neuroscience : the official journal of the Society for Neuroscience 2018 Sep 26;38(39):8484-8495
CD38-NAD(+)Axis Regulates Immunotherapeutic Anti-Tumor T Cell Response.
Chatterjee S, Daenthanasanmak A, Chakraborty P, Wyatt MW, Dhar P, Selvam SP, Fu J, Zhang J, Nguyen H, Kang I, Toth K, Al-Homrani M, Husain M, Beeson G, Ball L, Helke K, Husain S, Garrett-Mayer E, Hardiman G, Mehrotra M, Nishimura MI, Beeson CC, Bupp MG, Wu J, Ogretmen B, Paulos CM, Rathmell J, Yu XZ, Mehrotra S
Cell metabolism 2018 Jan 9;27(1):85-100.e8
Cell metabolism 2018 Jan 9;27(1):85-100.e8
Initiation of Antiviral B Cell Immunity Relies on Innate Signals from Spatially Positioned NKT Cells.
Gaya M, Barral P, Burbage M, Aggarwal S, Montaner B, Warren Navia A, Aid M, Tsui C, Maldonado P, Nair U, Ghneim K, Fallon PG, Sekaly RP, Barouch DH, Shalek AK, Bruckbauer A, Strid J, Batista FD
Cell 2018 Jan 25;172(3):517-533.e20
Cell 2018 Jan 25;172(3):517-533.e20
Regulation of chitinase-3-like-1 in T cell elicits Th1 and cytotoxic responses to inhibit lung metastasis.
Kim DH, Park HJ, Lim S, Koo JH, Lee HG, Choi JO, Oh JH, Ha SJ, Kang MJ, Lee CM, Lee CG, Elias JA, Choi JM
Nature communications 2018 Feb 5;9(1):503
Nature communications 2018 Feb 5;9(1):503
Conventional NK cells and ILC1 are partially ablated in the livers of Ncr1 (iCre)Tbx21 (fl/fl) mice.
Cuff AO, Male V
Wellcome open research 2017;2:39
Wellcome open research 2017;2:39
Antibiotic-induced perturbations in microbial diversity during post-natal development alters amyloid pathology in an aged APP(SWE)/PS1(ΔE9) murine model of Alzheimer's disease.
Minter MR, Hinterleitner R, Meisel M, Zhang C, Leone V, Zhang X, Oyler-Castrillo P, Zhang X, Musch MW, Shen X, Jabri B, Chang EB, Tanzi RE, Sisodia SS
Scientific reports 2017 Sep 5;7(1):10411
Scientific reports 2017 Sep 5;7(1):10411
Resistance to TGFβ suppression and improved anti-tumor responses in CD8(+) T cells lacking PTPN22.
Brownlie RJ, Garcia C, Ravasz M, Zehn D, Salmond RJ, Zamoyska R
Nature communications 2017 Nov 7;8(1):1343
Nature communications 2017 Nov 7;8(1):1343
The NLRP3 inflammasome contributes to host protection during Sporothrix schenckii infection.
Gonçalves AC, Ferreira LS, Manente FA, de Faria CMQG, Polesi MC, de Andrade CR, Zamboni DS, Carlos IZ
Immunology 2017 Jun;151(2):154-166
Immunology 2017 Jun;151(2):154-166
CD36 receptor regulates malaria-induced immune responses primarily at early blood stage infection contributing to parasitemia control and resistance to mortality.
Thylur RP, Wu X, Gowda NM, Punnath K, Neelgund SE, Febbraio M, Gowda DC
The Journal of biological chemistry 2017 Jun 2;292(22):9394-9408
The Journal of biological chemistry 2017 Jun 2;292(22):9394-9408
B-cell anergy induces a Th17 shift in a novel B lymphocyte transgenic NOD mouse model, the 116C-NOD mouse.
Carrascal J, Carrillo J, Arpa B, Egia-Mendikute L, Rosell-Mases E, Pujol-Autonell I, Planas R, Mora C, Mauricio D, Ampudia RM, Vives-Pi M, Verdaguer J
European journal of immunology 2016 Mar;46(3):593-608
European journal of immunology 2016 Mar;46(3):593-608
CXCL16-positive dendritic cells enhance invariant natural killer T cell-dependent IFNγ production and tumor control.
Veinotte L, Gebremeskel S, Johnston B
Oncoimmunology 2016 Jun;5(6):e1160979
Oncoimmunology 2016 Jun;5(6):e1160979
IL12-mediated sensitizing of T-cell receptor-dependent and -independent tumor cell killing.
Braun M, Ress ML, Yoo YE, Scholz CJ, Eyrich M, Schlegel PG, Wölfl M
Oncoimmunology 2016 Jul;5(7):e1188245
Oncoimmunology 2016 Jul;5(7):e1188245
Human Head and Neck Squamous Cell Carcinoma-Associated Semaphorin 4D Induces Expansion of Myeloid-Derived Suppressor Cells.
Younis RH, Han KL, Webb TJ
Journal of immunology (Baltimore, Md. : 1950) 2016 Feb 1;196(3):1419-29
Journal of immunology (Baltimore, Md. : 1950) 2016 Feb 1;196(3):1419-29
Oxygen Sensing by T Cells Establishes an Immunologically Tolerant Metastatic Niche.
Clever D, Roychoudhuri R, Constantinides MG, Askenase MH, Sukumar M, Klebanoff CA, Eil RL, Hickman HD, Yu Z, Pan JH, Palmer DC, Phan AT, Goulding J, Gattinoni L, Goldrath AW, Belkaid Y, Restifo NP
Cell 2016 Aug 25;166(5):1117-1131.e14
Cell 2016 Aug 25;166(5):1117-1131.e14
Intranasal Administration of Lentiviral miR-135a Regulates Mast Cell and Allergen-Induced Inflammation by Targeting GATA-3.
Deng YQ, Yang YQ, Wang SB, Li F, Liu MZ, Hua QQ, Tao ZZ
PloS one 2015;10(9):e0139322
PloS one 2015;10(9):e0139322
Chronic Trichuris muris Infection Decreases Diversity of the Intestinal Microbiota and Concomitantly Increases the Abundance of Lactobacilli.
Holm JB, Sorobetea D, Kiilerich P, Ramayo-Caldas Y, Estellé J, Ma T, Madsen L, Kristiansen K, Svensson-Frej M
PloS one 2015;10(5):e0125495
PloS one 2015;10(5):e0125495
Eliminating roles for T-bet and IL-2 but revealing superior activation and proliferation as mechanisms underpinning dominance of regulatory T cells in tumors.
Colbeck EJ, Hindley JP, Smart K, Jones E, Bloom A, Bridgeman H, McPherson RC, Turner DG, Ladell K, Price DA, O'Connor RA, Anderton SM, Godkin AJ, Gallimore AM
Oncotarget 2015 Sep 22;6(28):24649-59
Oncotarget 2015 Sep 22;6(28):24649-59
Eliminating roles for T-bet and IL-2 but revealing superior activation and proliferation as mechanisms underpinning dominance of regulatory T cells in tumors.
Colbeck EJ, Hindley JP, Smart K, Jones E, Bloom A, Bridgeman H, McPherson RC, Turner DG, Ladell K, Price DA, O'Connor RA, Anderton SM, Godkin AJ, Gallimore AM
Oncotarget 2015 Sep 22;6(28):24649-59
Oncotarget 2015 Sep 22;6(28):24649-59
IL-10 inhibits neuraminidase-activated TGF-β and facilitates Th1 phenotype during early phase of infection.
Dutta A, Huang CT, Chen TC, Lin CY, Chiu CH, Lin YC, Chang CS, He YC
Nature communications 2015 Mar 2;6:6374
Nature communications 2015 Mar 2;6:6374
Expression of GM-CSF in T Cells Is Increased in Multiple Sclerosis and Suppressed by IFN-β Therapy.
Rasouli J, Ciric B, Imitola J, Gonnella P, Hwang D, Mahajan K, Mari ER, Safavi F, Leist TP, Zhang GX, Rostami A
Journal of immunology (Baltimore, Md. : 1950) 2015 Jun 1;194(11):5085-93
Journal of immunology (Baltimore, Md. : 1950) 2015 Jun 1;194(11):5085-93
The MAPK-Activated Kinase MK2 Attenuates Dendritic Cell-Mediated Th1 Differentiation and Autoimmune Encephalomyelitis.
Soukup K, Halfmann A, Le Bras M, Sahin E, Vittori S, Poyer F, Schuh C, Luger R, Niederreiter B, Haider T, Stoiber D, Blüml S, Schabbauer G, Kotlyarov A, Gaestel M, Felzmann T, Dohnal AM
Journal of immunology (Baltimore, Md. : 1950) 2015 Jul 15;195(2):541-52
Journal of immunology (Baltimore, Md. : 1950) 2015 Jul 15;195(2):541-52
The transduction pattern of IL-12-encoding lentiviral vectors shapes the immunological outcome.
Goyvaerts C, Broos K, Escors D, Heirman C, Raes G, De Baetselier P, Thielemans K, Breckpot K
European journal of immunology 2015 Dec;45(12):3351-61
European journal of immunology 2015 Dec;45(12):3351-61
T cell factor-1 controls the lifetime of CD4+ CD8+ thymocytes in vivo and distal T cell receptor α-chain rearrangement required for NKT cell development.
Sharma A, Berga-Bolaños R, Sen JM
PloS one 2014;9(12):e115803
PloS one 2014;9(12):e115803
Prophylaxis with a respiratory syncytial virus (RSV) anti-G protein monoclonal antibody shifts the adaptive immune response to RSV rA2-line19F infection from Th2 to Th1 in BALB/c mice.
Boyoglu-Barnum S, Chirkova T, Todd SO, Barnum TR, Gaston KA, Jorquera P, Haynes LM, Tripp RA, Moore ML, Anderson LJ
Journal of virology 2014 Sep;88(18):10569-83
Journal of virology 2014 Sep;88(18):10569-83
High eomesodermin expression among CD57+ CD8+ T cells identifies a CD8+ T cell subset associated with viral control during chronic human immunodeficiency virus infection.
Simonetta F, Hua S, Lécuroux C, Gérard S, Boufassa F, Sáez-Cirión A, Pancino G, Goujard C, Lambotte O, Venet A, Bourgeois C
Journal of virology 2014 Oct;88(20):11861-71
Journal of virology 2014 Oct;88(20):11861-71
MicroRNA-155 is essential for T(H)2-mediated allergen-induced eosinophilic inflammation in the lung.
Malmhäll C, Alawieh S, Lu Y, Sjöstrand M, Bossios A, Eldh M, Rådinger M
The Journal of allergy and clinical immunology 2014 May;133(5):1429-38, 1438.e1-7
The Journal of allergy and clinical immunology 2014 May;133(5):1429-38, 1438.e1-7
CD4CD8αα lymphocytes, a novel human regulatory T cell subset induced by colonic bacteria and deficient in patients with inflammatory bowel disease.
Sarrabayrouse G, Bossard C, Chauvin JM, Jarry A, Meurette G, Quévrain E, Bridonneau C, Preisser L, Asehnoune K, Labarrière N, Altare F, Sokol H, Jotereau F
PLoS biology 2014 Apr;12(4):e1001833
PLoS biology 2014 Apr;12(4):e1001833
Alterations in regulatory T cells induced by specific oligosaccharides improve vaccine responsiveness in mice.
Schijf MA, Kerperien J, Bastiaans J, Szklany K, Meerding J, Hofman G, Boon L, van Wijk F, Garssen J, Van't Land B
PloS one 2013;8(9):e75148
PloS one 2013;8(9):e75148
Proinflammatory Vδ2+ T cells populate the human intestinal mucosa and enhance IFN-γ production by colonic αβ T cells.
McCarthy NE, Bashir Z, Vossenkämper A, Hedin CR, Giles EM, Bhattacharjee S, Brown SG, Sanders TJ, Whelan K, MacDonald TT, Lindsay JO, Stagg AJ
Journal of immunology (Baltimore, Md. : 1950) 2013 Sep 1;191(5):2752-63
Journal of immunology (Baltimore, Md. : 1950) 2013 Sep 1;191(5):2752-63
Helicobacter pylori infection in a pig model is dominated by Th1 and cytotoxic CD8+ T cell responses.
Kronsteiner B, Bassaganya-Riera J, Philipson C, Viladomiu M, Carbo A, Pedragosa M, Vento S, Hontecillas R
Infection and immunity 2013 Oct;81(10):3803-13
Infection and immunity 2013 Oct;81(10):3803-13
Systemic 4-1BB activation induces a novel T cell phenotype driven by high expression of Eomesodermin.
Curran MA, Geiger TL, Montalvo W, Kim M, Reiner SL, Al-Shamkhani A, Sun JC, Allison JP
The Journal of experimental medicine 2013 Apr 8;210(4):743-55
The Journal of experimental medicine 2013 Apr 8;210(4):743-55
Breakdown in repression of IFN-γ mRNA leads to accumulation of self-reactive effector CD8+ T cells.
Chang PP, Lee SK, Hu X, Davey G, Duan G, Cho JH, Karupiah G, Sprent J, Heath WR, Bertram EM, Vinuesa CG
Journal of immunology (Baltimore, Md. : 1950) 2012 Jul 15;189(2):701-10
Journal of immunology (Baltimore, Md. : 1950) 2012 Jul 15;189(2):701-10
Increase in Th17 and T-reg lymphocytes and decrease of IL22 correlate with the recovery phase of acute EAE in rat.
Almolda B, Costa M, Montoya M, González B, Castellano B
PloS one 2011;6(11):e27473
PloS one 2011;6(11):e27473
Differential requirement for the CD45 splicing regulator hnRNPLL for accumulation of NKT and conventional T cells.
Yabas M, Godfrey DI, Goodnow CC, Hoyne GF
PloS one 2011;6(11):e26440
PloS one 2011;6(11):e26440
DOCK8 deficiency impairs CD8 T cell survival and function in humans and mice.
Randall KL, Chan SS, Ma CS, Fung I, Mei Y, Yabas M, Tan A, Arkwright PD, Al Suwairi W, Lugo Reyes SO, Yamazaki-Nakashimada MA, Garcia-Cruz Mde L, Smart JM, Picard C, Okada S, Jouanguy E, Casanova JL, Lambe T, Cornall RJ, Russell S, Oliaro J, Tangye SG, Bertram EM, Goodnow CC
The Journal of experimental medicine 2011 Oct 24;208(11):2305-20
The Journal of experimental medicine 2011 Oct 24;208(11):2305-20
CD134 plus CD137 dual costimulation induces Eomesodermin in CD4 T cells to program cytotoxic Th1 differentiation.
Qui HZ, Hagymasi AT, Bandyopadhyay S, St Rose MC, Ramanarasimhaiah R, Ménoret A, Mittler RS, Gordon SM, Reiner SL, Vella AT, Adler AJ
Journal of immunology (Baltimore, Md. : 1950) 2011 Oct 1;187(7):3555-64
Journal of immunology (Baltimore, Md. : 1950) 2011 Oct 1;187(7):3555-64
Cytokine requirements for the differentiation and expansion of IL-17A- and IL-22-producing human Vgamma2Vdelta2 T cells.
Ness-Schwickerath KJ, Jin C, Morita CT
Journal of immunology (Baltimore, Md. : 1950) 2010 Jun 15;184(12):7268-80
Journal of immunology (Baltimore, Md. : 1950) 2010 Jun 15;184(12):7268-80
Differential expression of T-bet, a T-box transcription factor required for Th1 T-cell development, in peripheral T-cell lymphomas.
Dorfman DM, van den Elzen P, Weng AP, Shahsafaei A, Glimcher LH
American journal of clinical pathology 2003 Dec;120(6):866-73
American journal of clinical pathology 2003 Dec;120(6):866-73
A novel transcription factor, T-bet, directs Th1 lineage commitment.
Szabo SJ, Kim ST, Costa GL, Zhang X, Fathman CG, Glimcher LH
Cell 2000 Mar 17;100(6):655-69
Cell 2000 Mar 17;100(6):655-69
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- Intracellular staining of normal human peripheral blood cells with Anti-Human CD8a APC (Product # 17-0086-42) and 0.125 µg of Anti-Human/Mouse T-bet PerCP-Cyanine5-5 using the Foxp3 Fixation/Permeabilization Buffers (Product # 00-5523-00). Cells in the lymphocyte gate were used for analysis.
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- Figure 2 1 wk ABX gvg-treated APP SWE /PS1 DeltaE9 mice display altered peripheral and brain inflammatory profiles. ( A ) Representative density dot plots of T-bet and Foxp3 intracellular expression in TCRbeta + CD4 + T cell populations isolated from MLN, blood and brain tissue of vehicle and 1 wk ABX gvg-treated APP SWE /PS1 DeltaE9 mice analysed by flow cytometry. Quantified percentages of ( B ) Foxp3 + and ( C ) T-bet + CD4 + T cells, representative of a T-reg and Th1 T cell phenotype respectively, are expressed relative to total live CD4 + T cell counts ( n = 5-6, *p < 0.05, **p < 0.01, un-paired two-tailed Student's t -test). ( D ) Immunoblot-based array of inflammatory mediators present in the serum of vehicle and 1 wk ABX gvg-treated APP SWE /PS1 DeltaE9 mice ( n = 10 pooled sera). ( E ) Immunoblot-based array of inflammatory mediators present in the CSF of vehicle and 1 wk ABX gvg-treated APP SWE /PS1 DeltaE9 mice ( n = 10 pooled CSF). ( F ) Heat map analysis of inflammatory mediator fold change expression in 1 wk ABX gvg-treated APP SWE /PS1 DeltaE9 mice relative to control. Data are displayed as log e (mean) or mean +- SEM. See Supp. Figs 2 , 4 , 5 , 6 , and statistical Table 2 for additional information.
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- Fig 5 Lentiviral-mmu-miR-135a treatment influences Th cell polarization. The expression of T-bet and GATA-3 protein in CD4 + T cells was measured in the spleens of normal (control), AR (AR-induced), positive (AR-induced, treated with lentiviral-mmu-miR-135a), and negative (AR-induced, treated with empty lentivirus) mice using flow cytometry. (A) Representative dot plots from each experimental group. The percentages of CD4 + T-bet + T cells (B) and CD4 + GATA-3 + T cells (C) were also calculated. Data are presented as the mean +- SEM. *P
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- Figure 7. Melanoma Patients Demonstrated an Overall Increase of PS Expression on Immune Subsets in the Blood 4-7 Days after RT Peripheral blood was collected from 7 melanoma patients before and 4-7 days after receiving tumor-directed RT. Freshly isolated PBMCs from each patient were stained for PS expression using annexin V on the day the blood was collected, as described in STAR Methods . (A) Histogram plots of annexin V staining of viable immune cell subsets in PBMCs from a single patient (Pt. 1) pretreatment (Tx) and post-RT. (B) Representative plots gated on live CD3+ CD8+ T cells of annexin V versus caspase-3/7 activity of PBMCs from a single patient pre-RT and 4 days post-RT. FMO, control. (C) Top: individual values for each patient. Bottom: average percentage +- SEM of annexin V+ immune cell subsets pre-RT and post-RT. *p < 0.05, ***p < 0.005. (D) Model summarizing the effects of targeting PS with RT and anti-PD-1 on immune cell activation.
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- Figure 4 Phenotype of artARENA-induced CTLs and their dependence on IL-33-ST2 alarmin signaling (A-F) We immunized BALB/c mice with artPICV-P1A and artLCMV-P1A in homologous or heterologous prime-boost vaccination i.v. on day 0 and day 27. On day 34, we analyzed P1A-Tet-binding and CD62L expression by splenic CD8 + T cells (A; gated on CD8 + B220 - lymphocytes). Unimmunized control mice are shown for comparison in (A) only. Numbers in (A) indicate the percentage of cells in the respective quadrant. Total P1A-Tet + CTLs (B), P1A-specific effector/effector memory CTLs (CD62L lo ; C), and P1A-specific central memory CTLs (CD62L hi ; D) were enumerated in the spleen on day 34. In both subsets of P1A-specific CTLs, CD62L hi and CD62L lo , we determined the surface expression of KLRG1, CX3CR1, CD27, CD43, and CD127 as well as the master transcription factors Tcf-1, Tbet, and Eomes (E). Total numbers of marker-expressing P1A-specific CTLs were enumerated in (F). (A) shows representative FACS plots from individual mice. Symbols in (B)-(D) and (F) represent individual mice, and bars in (B)-(D) and (F) indicate the mean +- SD. Numbers in (A) and (E) indicate the percentage of gated cells (mean +- SD) or the mean fluorescence intensity (MFI +- SD). Means were calculated from six mice per immunization group (A-F) or from three unimmunized controls (A). N = 2. **p < 0.01 by unpaired two-tailed Student''s t test. (G and H) We immunized ST2 -/- and WT mice with artLCMV-E7E6, artPICV-E7E6 or
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- Figure 1. Patients with Parkinson's disease (PD) and mice with experimental PD exhibit decreased regulatory T (Treg) and increased T helper 1 (Th1) cell numbers in the blood. Treg cells were defined as CD3+CD4+CD25+FoxP3+ cells, whereas Th1 cells were identified as CD3+CD4+T-bet+ cells. (a) Representative plots of Treg and Th1 cells in patients with PD and healthy volunteers (HVs). (b) Representative plots of Treg and Th1 cells in control mice (Saline) and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice. (c) Patients with PD exhibited lower Treg cell levels and greater Th1 cell levels in the blood than HVs. n = 20. ****, p < 0.0001 by a two-tailed Student's t tests. (d) MPTP-induced experimental PD mice had fewer Treg and more Th1 cells in the circulation than saline-treated mice. n = 9/group in each experiment performed in triplicate. ***, p < 0.001; ****, p < 0.0001, according to Student's t test.
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- Figure 1 Altered distribution of Th1, Th2, and Th17 in PBMCs of patients. (A) PBMCs were stained intracellularly with T-bet, GATA-3, and ROR-gammat mAbs after surface staining of CD4 mAb. According to CD4 staining and SSC, CD4 + T cells were gated. The parameters shown in quadrants of the representative graphs are mean frequency of GATA-3 for each group. (B-D) The average percentages of CD4 + T-bet + Th1, CD4 + GATA-3 + Th2, CD4 + ROR-gammat + Th17 are compared between HC (n = 24) and whole patients (n = 50) as well as subgroups of patients (SD, n = 24; LD, n = 26; FF, n = 33; LF, n = 17). Error bars represent mean+-SD. ** P < 0.01, * P < 0.05, and NS P >= 0.05.
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- Fig. 3 ADAM12 is essential for T-bet expression in T cells and IFNgamma production. a ADAM12 +/+ and ADAM12 -/- T cells were treated with plate-bound anti-CD3 (5 mug/ml) and anti-ADAM12 (10 mug/ml) for 3 days. Cell-culture supernatants were collected for ELISA. MOG 35-55 -specific T cells treated with MOG 35-55 (50 mug/ml), IL-12 (20 ng/ml), anti-IL-4 (10 mug/ml), and siRNAs (siControl or siADAM12) for 72 h. b FACS dot plots of the gating strategy and T-bet staining in CD4 + T cells. c Quantification of FACS results from b . d IFNgamma production in Th1 cell cultures by ELISA. Graphs in c, d are mean +- SEM from three independent experiments. *** P < 0.001 by Student's t test. Th17 cells polarized in vitro. e , f Representative FACS plots of gating strategy and T-bet- and RORgammat-stained CD4 + T cells. g Quantification of FACS results from a representative experiment--i.e., from two independent experiments. Graphs are mean +- SEM, N = 3. h IFNgamma and IL-17 production in Th17 cell cultures, by ELISA from three independent experiments. ** P < 0.01 by two-way ANOVA with post-Tukey's multiple comparisons test
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- Figure 13 Pa MR1T cell antigen-specific proliferation assay (A) Representative FACS plots showing frequencies of Pa MR1T cells (defined as CD3 + hMR1-5-OP-RU + ) and CTV dilution on day 0 and day 5 of culture with 5-OP-RU from a single Pa donor. (B) Representative FACS plots showing the expression levels of different transcription factors (PLZF, T-bet, Eomes, and RORgammat) and Perforin against CTV at the two different timepoints. Figure adapted and reprinted with permission from ().
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- Fig. 4 MAIT cells were polarized to functional MAIT-1 phenotype upon F. tularensis infection. A Representative flow cytometry plots showing intranuclear staining for T-bet (representing Th1) and RORgammat (Th17) in gated MAIT cells from the liver, lungs, spleen, kidneys, LPL, IEL, and blood of naive and infected mice on 6 and 40+ dpi (41 and 68 dpi in two experiments) with 10 4 CFU F. tularensis LVS i.v. Numbers in quadrants represent cell percentage. It is noteworthy that IEL from naive mice yielded insufficient numbers of MAIT cells for accurate assessment of transcription factor expression and, thus, were omitted from our analysis. B Percentage of MAIT cells expressing combinations of T-bet and RORgammat from the same mice in A . Pooled data from two independent experiments (mean +- SEM, n = 3-6 mice per group, as indicated). One-way ANOVA with Tukey's multiple comparisons test was performed on MAIT-1% in each organ (except for IEL) between time points as indicated; p -values are indicated, nd; not determined. Source data are provided as a Source Data file.
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- Fig. 7 MAIT cell-mediated protection in immune-compromised mice requires IFNgamma, TNF, and GM-CSF. A MAIT cell percentage of alphabeta-T cells in the liver and representative FACS plot showing intranuclear staining for T-bet and RORgammat of MAIT cells from donor C57BL/6 mice vaccinated with CpG and 5-OP-RU i.v. for 7 days, prior cell sorting for adoptive cell transfer. Pooled data from 7 (nil) or 11 (vaccinated) mice from 3 independent experiments (mean +- SEM). Unpaired t -test (two-tailed). P < 0.0001. B Schematic of protocol for MAIT cell adoptive transfer and F. tularensis LVS challenge: 10 5 liver MAIT cells from C57BL/6 (WT, shown in A ), Ifngamma -/- , Tnf -/- , Gm-csf -/- , or Il-17 -/- mice vaccinated with CpG (10 nmol) and 5-OP-RU (2 nmol) i.v. for 7 days were sorted by flow cytometry and transferred i.v. into Rag2 -/- gammaC -/- mice. The mice were treated with anti-CD4 and anti-CD8 mAb injection (i.p., 0.1 mg each) at days 1 and 3 post MAIT cell transfer, to deplete contaminating conventional T cells. After 2 weeks, mice were infected with an otherwise lethal dose (20 CFU) of F. tularensis LVS i.v. C Survival of untreated Rag2 -/- gammaC -/- mice or Rag2 -/- gammaC -/- mice following transfer of MAIT cells from WT, Ifngamma -/- , Tnf -/- , Gm-csf -/- , or Il-17 -/- mice according to schematic shown in B . Pooled data from two independent experiments with similar results ( n = 12-24 mice per group, as indicated). Log-rank tests ( Ifngamma -/- , Tnf -/- , Gm-csf -
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- Figure 3 Dynamics of Th1 cell population. A) Representative dot-plots of the population of CD4+Tbet+ cells of EAE animals. Dot-plots were obtained by previously gating in the CD3+ T cell population. Different quadrants were defined by application of the appropriate isotype control. A minimum of three animals per group was pooled and three replicates per score were analyzed. B and C) Histogrammes showing, respectively, the values corresponding to the total number and the percentage of CD4+Tbet+ cell population along EAE. Note that CD4+Tbet+ lymphocytes are found during the induction and peak phases and markedly decreased at score 2R of the recovery phase (ANOVA and Tukey's post-hoc test, *p
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- Figure 1 Homozygous RhoA deletion in Treg cells leads to early, fatal spontaneous inflammatory disorders. (A) Survival outcome of RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/Flox Foxp3 YFP-Cre mice. Results were analyzed with a log-rank (Mantel-Cox) test and expressed as Kaplan-Meier survival curves. (B) Image of lymphadenopathy in RhoA Flox/Flox Foxp3 YFP-Cre mice. Inguinal lymph nodes are shown. (C) Images of H&E staining of the indicated organs from RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/Flox Foxp3 YFP-Cre mice (original magnification X 400). (D) Left, representative flow cytogram of CD44 and CD62L staining in CD4 + and CD8 + cells from the spleen of RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/Flox Foxp3 YFP-Cre mice. The numbers indicate percentages of CD44 + , CD44 + CD62L + , and CD62L + cells. Right, average percentages of CD44 + , CD44 + CD62L + , and CD62L + cells. (E) Left, representative flow cytogram of IL-17, IFN-gamma, and IL-4 staining in CD4 + Foxp3 - cells from the spleen of RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/Flox Foxp3 YFP-Cre mice. The numbers indicate percentages of IL-17 + , IFN-gamma + , and IL-4 + cells. Right, average percentages of IL-17 + , IFN-gamma + , and IL-4 + cells. (F) Left, representative flow cytogram of RORgammaT, T-bet and GATA3 staining in CD4 + Foxp3 - cells from the spleen of RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/Flox Foxp3 YFP-Cre mice. The numbers indicate percentages of RORgammaT + , T-bet + , and GATA3 + cells. Right, average percentages of RORgam
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- Figure 2 Homozygous RhoA deletion in Treg cells dampens Treg cell homeostasis and induces Treg cell plasticity. (A) Left, representative flow cytogram of Foxp3 staining in CD4 + cells from the spleen of RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/Flox Foxp3 YFP-Cre mice. The numbers indicate percentages of CD4 + Foxp3 + Treg cells. Middle, average percentages of CD4 + Foxp3 + Treg cells. Right, average numbers of CD4 + Foxp3 + Treg cells. (B) Treg cell proliferation. Percentages of CD4 + Foxp3 + Treg cells incorporated with BrdU are shown. (C) Treg cell apoptosis. The expression levels (MFI: Mean fluorescence intensity) of active caspase 3 in CD4 + Foxp3 + Treg cells are shown. (D) The expression levels of Foxp3 in Treg cells. (E) Left, representative flow cytogram of IL-17, IFN-gamma, and IL-4 staining in CD4 + Foxp3 + Treg cells. The numbers indicate percentages of IL-17 + , IFN-gamma + , and IL-4 + Treg cells. Right, average percentages of IL-17 + , IFN-gamma + , and IL-4 + Treg cells. (F) Left, representative flow cytogram of RORgammaT, T-bet and GATA3 staining in CD4 + Foxp3 + Treg cells. The numbers indicate percentages of RORgammaT + , T-bet + , and GATA3 + Treg cells. Right, average percentages of RORgammaT + , T-bet + , and GATA3 + Treg cells. (G) Left, representative histogram of the expression levels of CTLA-4, GITR and PD-1 in CD4 + Foxp3 + Treg cells. The numbers above the graphs indicate MFI. Right, average MFI of CTLA-4, GITR and PD-1 in CD4 + Foxp3 + Treg cells. n =
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- Figure 5 Heterozygous RhoA deletion in Treg cells induces Treg cell plasticity and increases CD4 + effector T cells but does not result in autoimmunity. (A) Body weight of RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/+ Foxp3 YFP-Cre mice. (B) Images of H&E staining of the indicated organs. (C) Representative flow cytogram of Foxp3 staining in CD4 + cells from the spleen of RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/+ Foxp3 YFP-Cre mice. The numbers indicate percentages of CD4 + Foxp3 + Treg cells. (D) Left, average percentages of CD4 + Foxp3 + Treg cells. Right, cell numbers of CD4 + Foxp3 + Treg cells. (E) Proliferation of Foxp3 + YFP + Treg cells from RhoA +/+ Foxp3 YFP-Cre/+ and RhoA Flox/+ Foxp3 YFP-Cre/+ female mice. Percentages of Foxp3 + YFP + Treg cells incorporated with BrdU are shown. (F) Apoptosis of Foxp3 + YFP + Treg cells from RhoA +/+ Foxp3 YFP-Cre/+ and RhoA Flox/+ Foxp3 YFP-Cre/+ female mice. The expression levels (MFI: mean fluorescence intensity) of active caspase 3 in Foxp3 + YFP + Treg cells are shown. (G) The expression levels of Foxp3 in Treg cells from RhoA +/+ Foxp3 YFP-Cre and RhoA Flox/+ Foxp3 YFP-Cre mice. (H) Representative flow cytogram of IFN-gamma, IL-17 and IL-4 staining in CD4 + Foxp3 + Treg cells. The numbers indicate percentages of IFN-gamma + , IL-17 + and IL-4 + Treg cells. (I) Average percentages of IFN-gamma + , IL-17 + and IL-4 + Treg cells. (J) Representative flow cytogram of RORgammaT, T-bet and GATA3 staining in CD4 + Foxp3 + Treg cells. The numb
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- Figure 2 Effector CD4 T Cell Response to Adjuvanted Vaccines Groups of C57BL/6 mice were vaccinated IN, as in Figure 1 . At day 8 PV, cells from lungs and BAL were stained with I-A b /NP311 tetramers along with antibodies to cell surface molecules and transcription factors. (A) FACS plots show the percentages of I-A b /NP311 tetramer-binding cells among CD4 T cells. (B) Percentages of the indicated cell population among NP311-specific, tetramer-binding CD4 T cells. (C) FACS plots are gated on I-A b /NP311 tetramer-binding cells, and the numbers in each quadrant are the percentages of cells among the gated population; MFIs for transcription factors in NP311-specific CD4 T cells are plotted in the adjoining graphs. (D) FACS plots in (C) were used to quantify the percentages of T-bet LO EOMES HI cells (quadrant 4) among NP311-specific CD4 T cells. (E) Percentages of CD103 HI and CD69 HI cells among NP311-specific CD4 T cells. Data are representative of two independent experiments. Comparisons were made using one-way ANOVA test with Tukey-corrected multiple comparisons; *p < 0.05, **p < 0.01, and ***p < 0.001.
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- Fig. 9 In vivo administration of dNP2-siChi3l1 complex inhibits pulmonary metastasis with enhanced Th1 and CTL effector molecules. a Experimental scheme of dNP2-siRNA complex treatment in pulmonary melanoma metastasis model. b Representative lung image of dNP2-siEGFP and dNP2-siChi3l1-treated mice. Scale bar, 2 mm c Number of pleural colonies in each lung was counted. Data are mean +- SEM of three sets of independent experiments and each dot in graphs represent an individual mouse. d Relative total tumor area in the lung was measured by Image J software 1.48 v. e Histology of lung sections by H&E staining, and infiltrated tumor region was measured by Image J software 1.48 v. Scale bar, 200 m f , g IFNgamma producing CD4 and CD8 T cells, and Granzyme B expression level in IFNgamma + CD8 T cells in the lung was analyzed by intracellular staining. % of IFNgamma + , and MFI of Granzyme B was represented as scattered graph. h Intracellular T-bet expression level in CD4 + IFNgamma + and CD8 + IFNgamma + population. MFI was represented as scattered graph. i mRNA expression of genes related to cytotoxicity and Th1 effector functions was analyzed by quantitative RT-PCR. Each gene expression level was normalized to beta-actin. Data are mean +- SD of three sets of independent experiments and each dot in graphs represent an individual mouse. n.s., not significant; * p < 0.05, ** p < 0.01, *** p < 0.001 (two-tailed Student's t -test)
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- Figure 2. Similarities between Infection-Induced Mouse FCRL5 + B Cells and atMBCs from Plasmodium -Infected Humans (A) RNA-seq was performed on class-switched (IgM - IgD - ), FCRL5 - or FCRL5 + B cells sorted from the blood of Ifngr1 - / - mice infected for 21 days with P. chabaudi . Columns represent individual mice (n = 5). Select genes involved in immune signaling are shown. Genes with putative activating function are labeled in green; those with putative inhibitory function are labeled in red. (B) Heatmaps depict all genes that are differentially regulated in both human atMBCs (left) and mouse FCRL5 + B cells, relative to cMBCs and FCRL5 - B cells, respectively. Human data are from . (C-E) Expression of (C) CD72 and CD38, (D) T-bet, and (E) CD11b, CD11c, CD86, and CD40 were measured by flow cytometry in blood B cells 21 d.p.i. **p < 0.01; ***p < 0.001 by paired t test (Wilcoxon rank-sum test). See also Figure S2 and Tables S1 and S2 .
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- Figure 1. Characterization of CD4 + TIL, dLN, and Arm Transcriptomes by scRNA-Seq (A-D) TILs and dLN cells from wild-type (WT) mice at day 14 after MC38-GP injection analyzed by scRNA-seq and flow cytometry. (A) Heatmap shows row-standardized expression of selected genes across TIL and dLN clusters. Bar plot indicates the percentage of cells in each cluster relative to the total TIL or dLN cell number. (B) Flow cytometry contour plots of Foxp3 versus T-bet in CD44 hi GP66 + dLN cells (left) and in CD44 hi CD4 + splenocytes from tumor-free control mice (right). (C) Flow cytometry contour plots of Foxp3 versus T-bet in PD-1 + and GP66 + TILs (left) and in CD44 hi CD4 + splenocytes from tumor-free control mice (right). (B and C) Data representative from 18 tumor-bearing mice analyzed in four separate experiments. (D) t-SNE display of TILs and dLN cells, shaded gray by tissue origin (left) or color coded by main group (right, as defined in A). (E) t-SNE (TIL and dLN cell positioning as shown in B) display of normalized expression levels of selected genes. (F) Heatmap shows Pearson correlation between cluster fold change vectors (as defined in the text) across the two replicate experiments for TILs (left) and dLN cells (right). See also Figures S1 and S2 and Tables S1 and S6 .
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- Figure 3 Characteristics of iKP-19-CAR-T cells and CD19-CAR-T cells. iKP-19-CAR-T cells or CD19-CAR-T cells were cultured for 10 days in X-VIVO media supplemented with 100 U/mL IL-2. ( A ) The expression of T cell differentiation markers in CAR-T cells was analyzed by flow cytometry using PE/Cy7-anti-human CCR7 antibody, PE-anti-human-CD45RO antibody and PE-anti-human GzmB antibody ( n = 4 different donors). ( B ) The frequency of naive (T N ; CCR7 + CD45RA + ), T CM (CCR7 + CD45RA - ), effector memory (T EM ; CCR7 - CD45RA - ) or effector (T E ; CCR7 - CD45RA + ) T cells were analyzed by flow cytometry using PE/Cy7-anti-human CCR7 antibody and FITC-anti-human CD45RA antibody ( n = 4 different donors). ( C ) The expression of transcription factor Eomes in CAR-T cells was analyzed by flow cytometry using FITC-anti-human Eomes antibody ( n = 4 different donors). ( D ) The expression of transcription factor T-bet in CAR-T cells was analyzed by flow cytometry using PE-anti-human T-bet antibody ( n = 4 different donors). ( E ) The expression of T cell exhaustion marker PD-1 in CAR-T cells was analyzed by flow cytometry using FITC-anti-human PD-1 antibody ( n = 4 different donors). All experiments were performed in triplicate manner using PBMCs from each donor and MFI or percentage was statistically analyzed. * p < 0.05, ** p < 0.01, *** p < 0.001. Error bars represent +- SD.