Antibody data
- Antibody Data
- Antigen structure
- References [32]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [14]
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- Product number
- 15-1278-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD127 Monoclonal Antibody (eBioRDR5), PE-Cyanine5, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The eBioRDR5 monoclonal antibody reacts with human CD127 (Interleukin-7 Receptor alpha). CD127 complexes with CD132, also known as the common gamma chain (gamma c), to form the multi-functional IL-7 receptor (IL-7R). CD127 is a type I glycoprotein with a molecular weight of 75-80 kDa and is expressed by immature B cells through the early pre-B stage, by thymocytes during several stages of their development, and on most mature T cells, with transient down-regulation upon activation. Binding of IL-7 results in signal transduction which occurs through several tyrosine kinase pathways including the Jak/STAT pathway. IL-7 is indispensible for the development of lymphocytes, and the control of homeostatic proliferation of T-cells in the periphery. In addition, IL-7R signaling is know to be involved in the regulation of T cell receptor (TCR) locus rearrangement in gamma delta T cells.
- Antibody clone number
- eBioRDR5
- Concentration
- 5 µL/Test
Submitted references Baricitinib treatment resolves lower-airway macrophage inflammation and neutrophil recruitment in SARS-CoV-2-infected rhesus macaques.
Baricitinib treatment resolves lower airway inflammation and neutrophil recruitment in SARS-CoV-2-infected rhesus macaques.
CTLA-4 and PD-1 dual blockade induces SIV reactivation without control of rebound after antiretroviral therapy interruption.
The Effects of High Mobility Group Box-1 Protein on Peripheral Treg/Th17 Balance in Patients with Atherosclerosis.
Regulatory T cells as a biomarker for response to adalimumab in rheumatoid arthritis.
Activation of miR-21-Regulated Pathways in Immune Aging Selects against Signatures Characteristic of Memory T Cells.
A probiotic modulates the microbiome and immunity in multiple sclerosis.
Characterization of CD127(-) CD25(++) Treg from human colostrum.
ATP Release from Chemotherapy-Treated Dying Leukemia Cells Elicits an Immune Suppressive Effect by Increasing Regulatory T Cells and Tolerogenic Dendritic Cells.
Serial immunomonitoring of cancer patients receiving combined antagonistic anti-CD40 and chemotherapy reveals consistent and cyclical modulation of T cell and dendritic cell parameters.
HDAC inhibition potentiates immunotherapy in triple negative breast cancer.
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
Anti-TNF drives regulatory T cell expansion by paradoxically promoting membrane TNF-TNF-RII binding in rheumatoid arthritis.
The Distribution of Human Stem Cell-like Memory T Cell in Lung Cancer.
Efficacy, durability, and response predictors of low-dose interleukin-2 therapy for chronic graft-versus-host disease.
Decreased T Follicular Regulatory Cell/T Follicular Helper Cell (TFH) in Simian Immunodeficiency Virus-Infected Rhesus Macaques May Contribute to Accumulation of TFH in Chronic Infection.
Improved Natural Killer cell activity and retained anti-tumor CD8(+) T cell responses contribute to the induction of a pathological complete response in HER2-positive breast cancer patients undergoing neoadjuvant chemotherapy.
CMV-specific T cells generated from naïve T cells recognize atypical epitopes and may be protective in vivo.
Enhanced suppressor function of TIM-3+ FoxP3+ regulatory T cells.
Highly heterogeneous, activated, and short-lived regulatory T cells during chronic filarial infection.
The number of CCR5 expressing CD4+ T lymphocytes is lower in HIV-infected long-term non-progressors with viral control compared to normal progressors: a cross-sectional study.
Phase I study of alemtuzumab for therapy of steroid-refractory chronic graft-versus-host disease.
Bortezomib-based graft-versus-host disease prophylaxis in HLA-mismatched unrelated donor transplantation.
A different immunologic profile characterizes patients with HER-2-overexpressing and HER-2-negative locally advanced breast cancer: implications for immune-based therapies.
Bet v 1-specific T-cell receptor/forkhead box protein 3 transgenic T cells suppress Bet v 1-specific T-cell effector function in an activation-dependent manner.
Interleukin-2 and regulatory T cells in graft-versus-host disease.
Uncoupling of proliferation and cytokines from suppression within the CD4+CD25+Foxp3+ T-cell compartment in the 1st year of human type 1 diabetes.
Efficacious early antiviral activity of HIV Gag- and Pol-specific HLA-B 2705-restricted CD8+ T cells.
Fc receptor-like 3 protein expressed on IL-2 nonresponsive subset of human regulatory T cells.
Allosuppressive donor CD4+CD25+ regulatory T cells detach from the graft and circulate in recipients after liver transplantation.
Loss of IL-7 receptor alpha on CD4+ T cells defines terminally differentiated B cell-helping effector T cells in a B cell-rich lymphoid tissue.
Loss of IL-7 receptor alpha on CD4+ T cells defines terminally differentiated B cell-helping effector T cells in a B cell-rich lymphoid tissue.
Hoang TN, Pino M, Boddapati AK, Viox EG, Starke CE, Upadhyay AA, Gumber S, Nekorchuk M, Busman-Sahay K, Strongin Z, Harper JL, Tharp GK, Pellegrini KL, Kirejczyk S, Zandi K, Tao S, Horton TR, Beagle EN, Mahar EA, Lee MYH, Cohen J, Jean SM, Wood JS, Connor-Stroud F, Stammen RL, Delmas OM, Wang S, Cooney KA, Sayegh MN, Wang L, Filev PD, Weiskopf D, Silvestri G, Waggoner J, Piantadosi A, Kasturi SP, Al-Shakhshir H, Ribeiro SP, Sekaly RP, Levit RD, Estes JD, Vanderford TH, Schinazi RF, Bosinger SE, Paiardini M
Cell 2021 Jan 21;184(2):460-475.e21
Cell 2021 Jan 21;184(2):460-475.e21
Baricitinib treatment resolves lower airway inflammation and neutrophil recruitment in SARS-CoV-2-infected rhesus macaques.
Hoang TN, Pino M, Boddapati AK, Viox EG, Starke CE, Upadhyay AA, Gumber S, Busman-Sahay K, Strongin Z, Harper JL, Tharp GK, Pellegrini KL, Kirejczyk S, Zandi K, Tao S, Horton TR, Beagle EN, Mahar EA, Lee MY, Cohen J, Jean SM, Wood JS, Connor-Stroud F, Stammen RL, Delmas OM, Wang S, Cooney KA, Sayegh MN, Wang L, Weiskopf D, Filev PD, Waggoner J, Piantadosi A, Kasturi SP, Al-Shakhshir H, Ribeiro SP, Sekaly RP, Levit RD, Estes JD, Vanderford TH, Schinazi RF, Bosinger SE, Paiardini M
bioRxiv : the preprint server for biology 2020 Sep 16;
bioRxiv : the preprint server for biology 2020 Sep 16;
CTLA-4 and PD-1 dual blockade induces SIV reactivation without control of rebound after antiretroviral therapy interruption.
Harper J, Gordon S, Chan CN, Wang H, Lindemuth E, Galardi C, Falcinelli SD, Raines SLM, Read JL, Nguyen K, McGary CS, Nekorchuk M, Busman-Sahay K, Schawalder J, King C, Pino M, Micci L, Cervasi B, Jean S, Sanderson A, Johns B, Koblansky AA, Amrine-Madsen H, Lifson J, Margolis DM, Silvestri G, Bar KJ, Favre D, Estes JD, Paiardini M
Nature medicine 2020 Apr;26(4):519-528
Nature medicine 2020 Apr;26(4):519-528
The Effects of High Mobility Group Box-1 Protein on Peripheral Treg/Th17 Balance in Patients with Atherosclerosis.
Ding JW, Zhou T, Zheng XX, Wang XA, Tong XH, Luo CY, Zhang ZQ, Yu B
Acta Cardiologica Sinica 2018 Sep;34(5):399-408
Acta Cardiologica Sinica 2018 Sep;34(5):399-408
Regulatory T cells as a biomarker for response to adalimumab in rheumatoid arthritis.
Nguyen DX, Cotton A, Attipoe L, Ciurtin C, Doré CJ, Ehrenstein MR
The Journal of allergy and clinical immunology 2018 Sep;142(3):978-980.e9
The Journal of allergy and clinical immunology 2018 Sep;142(3):978-980.e9
Activation of miR-21-Regulated Pathways in Immune Aging Selects against Signatures Characteristic of Memory T Cells.
Kim C, Hu B, Jadhav RR, Jin J, Zhang H, Cavanagh MM, Akondy RS, Ahmed R, Weyand CM, Goronzy JJ
Cell reports 2018 Nov 20;25(8):2148-2162.e5
Cell reports 2018 Nov 20;25(8):2148-2162.e5
A probiotic modulates the microbiome and immunity in multiple sclerosis.
Tankou SK, Regev K, Healy BC, Tjon E, Laghi L, Cox LM, Kivisäkk P, Pierre IV, Hrishikesh L, Gandhi R, Cook S, Glanz B, Stankiewicz J, Weiner HL
Annals of neurology 2018 Jun;83(6):1147-1161
Annals of neurology 2018 Jun;83(6):1147-1161
Characterization of CD127(-) CD25(++) Treg from human colostrum.
Cérbulo-Vázquez A, Hernández-Peláez G, Arriaga-Pizano LA, Bautista-Pérez P, Romero-Venado J, Flores-González JC, Figueroa-Damian R, Soriano-Becerril D, Mancilla-Herrera I
American journal of reproductive immunology (New York, N.Y. : 1989) 2018 Feb;79(2)
American journal of reproductive immunology (New York, N.Y. : 1989) 2018 Feb;79(2)
ATP Release from Chemotherapy-Treated Dying Leukemia Cells Elicits an Immune Suppressive Effect by Increasing Regulatory T Cells and Tolerogenic Dendritic Cells.
Lecciso M, Ocadlikova D, Sangaletti S, Trabanelli S, De Marchi E, Orioli E, Pegoraro A, Portararo P, Jandus C, Bontadini A, Redavid A, Salvestrini V, Romero P, Colombo MP, Di Virgilio F, Cavo M, Adinolfi E, Curti A
Frontiers in immunology 2017;8:1918
Frontiers in immunology 2017;8:1918
Serial immunomonitoring of cancer patients receiving combined antagonistic anti-CD40 and chemotherapy reveals consistent and cyclical modulation of T cell and dendritic cell parameters.
McDonnell AM, Cook A, Robinson BWS, Lake RA, Nowak AK
BMC cancer 2017 Jun 15;17(1):417
BMC cancer 2017 Jun 15;17(1):417
HDAC inhibition potentiates immunotherapy in triple negative breast cancer.
Terranova-Barberio M, Thomas S, Ali N, Pawlowska N, Park J, Krings G, Rosenblum MD, Budillon A, Munster PN
Oncotarget 2017 Dec 26;8(69):114156-114172
Oncotarget 2017 Dec 26;8(69):114156-114172
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
Su S, Liao J, Liu J, Huang D, He C, Chen F, Yang L, Wu W, Chen J, Lin L, Zeng Y, Ouyang N, Cui X, Yao H, Su F, Huang JD, Lieberman J, Liu Q, Song E
Cell research 2017 Apr;27(4):461-482
Cell research 2017 Apr;27(4):461-482
Anti-TNF drives regulatory T cell expansion by paradoxically promoting membrane TNF-TNF-RII binding in rheumatoid arthritis.
Nguyen DX, Ehrenstein MR
The Journal of experimental medicine 2016 Jun 27;213(7):1241-53
The Journal of experimental medicine 2016 Jun 27;213(7):1241-53
The Distribution of Human Stem Cell-like Memory T Cell in Lung Cancer.
Hong H, Gu Y, Sheng SY, Lu CG, Zou JY, Wu CY
Journal of immunotherapy (Hagerstown, Md. : 1997) 2016 Jul-Aug;39(6):233-40
Journal of immunotherapy (Hagerstown, Md. : 1997) 2016 Jul-Aug;39(6):233-40
Efficacy, durability, and response predictors of low-dose interleukin-2 therapy for chronic graft-versus-host disease.
Koreth J, Kim HT, Jones KT, Lange PB, Reynolds CG, Chammas MJ, Dusenbury K, Whangbo J, Nikiforow S, Alyea EP 3rd, Armand P, Cutler CS, Ho VT, Chen YB, Avigan D, Blazar BR, Antin JH, Ritz J, Soiffer RJ
Blood 2016 Jul 7;128(1):130-7
Blood 2016 Jul 7;128(1):130-7
Decreased T Follicular Regulatory Cell/T Follicular Helper Cell (TFH) in Simian Immunodeficiency Virus-Infected Rhesus Macaques May Contribute to Accumulation of TFH in Chronic Infection.
Chowdhury A, Del Rio Estrada PM, Tharp GK, Trible RP, Amara RR, Chahroudi A, Reyes-Teran G, Bosinger SE, Silvestri G
Journal of immunology (Baltimore, Md. : 1950) 2015 Oct 1;195(7):3237-47
Journal of immunology (Baltimore, Md. : 1950) 2015 Oct 1;195(7):3237-47
Improved Natural Killer cell activity and retained anti-tumor CD8(+) T cell responses contribute to the induction of a pathological complete response in HER2-positive breast cancer patients undergoing neoadjuvant chemotherapy.
Muraro E, Comaro E, Talamini R, Turchet E, Miolo G, Scalone S, Militello L, Lombardi D, Spazzapan S, Perin T, Massarut S, Crivellari D, Dolcetti R, Martorelli D
Journal of translational medicine 2015 Jun 27;13:204
Journal of translational medicine 2015 Jun 27;13:204
CMV-specific T cells generated from naïve T cells recognize atypical epitopes and may be protective in vivo.
Hanley PJ, Melenhorst JJ, Nikiforow S, Scheinberg P, Blaney JW, Demmler-Harrison G, Cruz CR, Lam S, Krance RA, Leung KS, Martinez CA, Liu H, Douek DC, Heslop HE, Rooney CM, Shpall EJ, Barrett AJ, Rodgers JR, Bollard CM
Science translational medicine 2015 Apr 29;7(285):285ra63
Science translational medicine 2015 Apr 29;7(285):285ra63
Enhanced suppressor function of TIM-3+ FoxP3+ regulatory T cells.
Gautron AS, Dominguez-Villar M, de Marcken M, Hafler DA
European journal of immunology 2014 Sep;44(9):2703-2711
European journal of immunology 2014 Sep;44(9):2703-2711
Highly heterogeneous, activated, and short-lived regulatory T cells during chronic filarial infection.
Metenou S, Coulibaly YI, Sturdevant D, Dolo H, Diallo AA, Soumaoro L, Coulibaly ME, Kanakabandi K, Porcella SF, Klion AD, Nutman TB
European journal of immunology 2014 Jul;44(7):2036-47
European journal of immunology 2014 Jul;44(7):2036-47
The number of CCR5 expressing CD4+ T lymphocytes is lower in HIV-infected long-term non-progressors with viral control compared to normal progressors: a cross-sectional study.
Meijerink H, Indrati AR, van Crevel R, Joosten I, Koenen H, van der Ven AJ
BMC infectious diseases 2014 Dec 13;14:683
BMC infectious diseases 2014 Dec 13;14:683
Phase I study of alemtuzumab for therapy of steroid-refractory chronic graft-versus-host disease.
Nikiforow S, Kim HT, Bindra B, McDonough S, Glotzbecker B, Armand P, Koreth J, Ho VT, Alyea EP 3rd, Blazar BR, Ritz J, Soiffer RJ, Antin JH, Cutler CS
Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation 2013 May;19(5):804-11
Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation 2013 May;19(5):804-11
Bortezomib-based graft-versus-host disease prophylaxis in HLA-mismatched unrelated donor transplantation.
Koreth J, Stevenson KE, Kim HT, McDonough SM, Bindra B, Armand P, Ho VT, Cutler C, Blazar BR, Antin JH, Soiffer RJ, Ritz J, Alyea EP 3rd
Journal of clinical oncology : official journal of the American Society of Clinical Oncology 2012 Sep 10;30(26):3202-8
Journal of clinical oncology : official journal of the American Society of Clinical Oncology 2012 Sep 10;30(26):3202-8
A different immunologic profile characterizes patients with HER-2-overexpressing and HER-2-negative locally advanced breast cancer: implications for immune-based therapies.
Muraro E, Martorelli D, Turchet E, Miolo G, Scalone S, Comaro E, Talamini R, Mastorci K, Lombardi D, Perin T, Carbone A, Veronesi A, Crivellari D, Dolcetti R
Breast cancer research : BCR 2011;13(6):R117
Breast cancer research : BCR 2011;13(6):R117
Bet v 1-specific T-cell receptor/forkhead box protein 3 transgenic T cells suppress Bet v 1-specific T-cell effector function in an activation-dependent manner.
Schmetterer KG, Haiderer D, Leb-Reichl VM, Neunkirchner A, Jahn-Schmid B, Küng HJ, Schuch K, Steinberger P, Bohle B, Pickl WF
The Journal of allergy and clinical immunology 2011 Jan;127(1):238-45, 245.e1-3
The Journal of allergy and clinical immunology 2011 Jan;127(1):238-45, 245.e1-3
Interleukin-2 and regulatory T cells in graft-versus-host disease.
Koreth J, Matsuoka K, Kim HT, McDonough SM, Bindra B, Alyea EP 3rd, Armand P, Cutler C, Ho VT, Treister NS, Bienfang DC, Prasad S, Tzachanis D, Joyce RM, Avigan DE, Antin JH, Ritz J, Soiffer RJ
The New England journal of medicine 2011 Dec 1;365(22):2055-66
The New England journal of medicine 2011 Dec 1;365(22):2055-66
Uncoupling of proliferation and cytokines from suppression within the CD4+CD25+Foxp3+ T-cell compartment in the 1st year of human type 1 diabetes.
Hughson A, Bromberg I, Johnson B, Quataert S, Jospe N, Fowell DJ
Diabetes 2011 Aug;60(8):2125-33
Diabetes 2011 Aug;60(8):2125-33
Efficacious early antiviral activity of HIV Gag- and Pol-specific HLA-B 2705-restricted CD8+ T cells.
Payne RP, Kløverpris H, Sacha JB, Brumme Z, Brumme C, Buus S, Sims S, Hickling S, Riddell L, Chen F, Luzzi G, Edwards A, Phillips R, Prado JG, Goulder PJ
Journal of virology 2010 Oct;84(20):10543-57
Journal of virology 2010 Oct;84(20):10543-57
Fc receptor-like 3 protein expressed on IL-2 nonresponsive subset of human regulatory T cells.
Nagata S, Ise T, Pastan I
Journal of immunology (Baltimore, Md. : 1950) 2009 Jun 15;182(12):7518-26
Journal of immunology (Baltimore, Md. : 1950) 2009 Jun 15;182(12):7518-26
Allosuppressive donor CD4+CD25+ regulatory T cells detach from the graft and circulate in recipients after liver transplantation.
Demirkiran A, Bosma BM, Kok A, Baan CC, Metselaar HJ, Ijzermans JN, Tilanus HW, Kwekkeboom J, van der Laan LJ
Journal of immunology (Baltimore, Md. : 1950) 2007 May 15;178(10):6066-72
Journal of immunology (Baltimore, Md. : 1950) 2007 May 15;178(10):6066-72
Loss of IL-7 receptor alpha on CD4+ T cells defines terminally differentiated B cell-helping effector T cells in a B cell-rich lymphoid tissue.
Lim HW, Kim CH
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 1;179(11):7448-56
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 1;179(11):7448-56
Loss of IL-7 receptor alpha on CD4+ T cells defines terminally differentiated B cell-helping effector T cells in a B cell-rich lymphoid tissue.
Lim HW, Kim CH
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 1;179(11):7448-56
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 1;179(11):7448-56
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Supportive validation
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- Staining of normal human peripheral blood cells with Anti-Human CD3 FITC (Product # 11-0037-42) and Mouse IgG1 K Isotype Control PE-Cyanine5 (Product # 15-4714-81) (left) or Anti-Human CD127 PE-Cyanine5 (right).Cells in the lymphocyte gate were used for analysis.
Supportive validation
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- FIGURE 1 Identification of Tscm CD4 + CD45RA + CD45RO - CD62L + CCR7 + CD127 + CD27 + CD28 + CD95 + CD122 + T (Tscm) cell in human blood and lymph nodes. PBMCs were isolated from the blood of non-small cell lung cancer (NSCLC) patients (n=15) (NSCLC-PBMC) and healthy donors (n=11) (HD-PBMC); lymphocytes were isolated from the tumor-infiltrated lymph node of NSCLC patients who were collected blood at same time (n=7) (NSCLC-Ly); lymphocytes were isolated from the healthy lymph node of non lung cancer patients (n=7) (Normal-Ly), analyzed by flow cytometry. A, Representative flow cytometric analyses of CD4 + CD45RA + CD45RO - CD62L + CCR7 + CD127 + CD27 + CD28 + CD95 + CD122 + T cells, indicating Tscm cells. B, The frequency of the CD4 + Tscm cells in the HD-PBMC, NSCLC-PBMC, Normal-Ly, NSCLC-Ly. The events of CD4 + Tscm cells in the blood and lymph node from NSCLC patients and healthy donors, expressed as the mean+-SEM. C, The frequency of the CD8 + Tscm cells in the HD-PBMC, NSCLC-PBMC, Normal-Ly, NSCLC-Ly, expressed as the mean+-SEM. D, The events of Tscm of CD4 + and CD8 + cells in the blood and the lymph node from NSCLC patients and healthy donors. HD indicates healthy donors; IFN, interferon; PBMC, peripheral blood mononuclear cells; Tscm cell, stem cell-like memory T cell. * P
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- FIGURE 3 CD4 + CD45RA + CD45RO - CD95 - CD122 + CD127 + T cell population displays different phenotypes in human blood and lymph nodes. A, Flow cytometry plots showing IFN-gamma expression in the CD4 + CD45RA + /CD45RO - CD95 - CD122 + CD127 + T cells from the blood and lymph node of the non-small cell lung cancer (NSCLC) patients and healthy donors. B, The mean frequency (+-SEM) of the events of CD4 + CD45RA + /CD45RO - CD95 - CD122 + CD127 + T cells. C, The mean IFN-gamma production (+-SEM) of the CD4 + CD45RA + /CD45RO - CD95 - CD122 + CD127 + T cells. HD indicates healthy donors; IFN, interferon; PBMC, peripheral blood mononuclear cells; PMA, phorbol 12-myristate13-acetate. * P
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- Fig. 6 a Representative flow cytometry data, demonstrating the gating strategy used on PBMC for Treg identification and analysis. FSC-area vs. FSC-height was used for doublet discrimination. A ""dump"" channel was used to gate out dead cells (LIVE/DEAD fixable viability stain), CD14 + monocytes and CD19 + B cells, and lymphocytes were subsequently selected by FSC vs. SSC. CD4 + T cells were gated on the basis of CD4 vs. CD3 staining, then examined for expression of Ki67 and ICOS. Tregs were identified within the CD4 + T cell population as CD25 hi CD127 lo and Foxp3 + . b Longitudinal empirical data, linear mixed models and estimated means (left, centre and right-hand panels respectively) for Ki67+ and ICOS+ expression on CD4+ T cells, and the Treg proportion of CD4 cells ( P -values: *
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- Figure 6 In vivo knockdown of PITPNM3 in CD4 + T cells reverses immunosuppression and inhibits tumor progression in humanized mice. (A) Humanized mice bearing palpable MDA-MB-231 orthotopic xenografts were intraperitoneally injected daily for 14 days with PBS, 1 nmol CD4-aptamer-control siRNA (AsiC-con) or CD4-aptamer-siRNA targeting PITPNM3 (sequence in A , AsiC-PI) to assess the role of PITPNM3 in TI Tregs, and other T cells and tumor control. Experimental schematic is provided in Supplementary information, Figure S9A . (B) Representative immunoblots showing selective knockdown of PITPNM3 protein in PB CD4 + T cells, but not tumor xenografts ( n = 3). (C) PITPNM3 knockdown did not affect the distribution of human CD45 + hematopoietic cells, CD4 + and CD8 + T cells, and CD14 + monocytes in the peripheral blood of humanized mice. Representative flow plots are shown ( n = 3). (D , E) Effect of PITPNM3 knockdown on TI naive CD4 + , Tregs and CD8 + T cell numbers, and apoptosis by TUNEL assay in xenografts. D shows representative immunofluorescence microscopy images. Top row indicates CD4 + naive T cells by arrows; the second row indicates CD4 + CD45RO + Foxp3 - CD4 + memory T cells (yellow arrows) and Foxp3 + Tregs (white arrows). Scale bar, 50 mum. E shows number of cells of each subtype/high power field in eight mice ( ** P < 0.01, *** P < 0.001 compared to PBS group by Student's t -test). (F) Flow cytometry analysis of gated human CD3 + CD4 + cells isolated from xenogra
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- Figure 4. miR-21 Promotes Effector Cell Differentiation through AP-1 Activation (A) Naive CD4 + T cells were transfected with either scrambled control or miR-21-blocking locked nucleic acid (LNA21). After 48 hr, PDCD4 and b-actin expression were assessed by western blot. Representative blots and mean normalized intensities from four experiments are shown (mean +- SEM, two-tailed paired t test). (B) Naive CD4 + T cells were activated with anti-CD3 and anti-CD28 beads and transduced with a lentiviral vector expressing scrambled control or anti-miR-21. The representative histogram shows phosphorylated JNK in GFP + cells on day 3. The filled gray histogram represents unstimulated naive CD4 + T cells. Results from 7 experiments are expressed relative to the geometric mean fluorescence intensity (MFI) of controls (mean +- SEM, two-tailed paired t test). (C) Naive CD4 + T cells were activated and transduced as described in (B) and co-transfected with the AP-1 luciferase reporter plasmid and the Renilla luciferase control construct. On day 3, the activity of AP-1 firefly luciferase was measured and normalized to that of Renilla luciferase (n = 4, mean +- SEM, two-tailed paired t test). (D-F) Naive CD4 + T cells isolated from 20- to 35-year-old and 65- to 85-year-old individuals were activated with beads coated with anti-CD3 and anti-CD28 antibodies. (D) PDCD4 expression was quantified by RT-PCR on day 3 and day 5. Results are normalized to ACTB and presented relative to