Antibody data
- Antibody Data
- Antigen structure
- References [44]
- Comments [0]
- Validations
- Immunohistochemistry [1]
- Flow cytometry [2]
- Other assay [11]
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- Product number
- MA5-14548 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD8 Monoclonal Antibody (SP16)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- MA5-14548 targets CD8 in IHC (P), and FACS applications and shows reactivity with Human samples.
- Antibody clone number
- SP16
- Concentration
- 0.022 mg/mL
Submitted references Single-cell transcriptomics reveals distinct effector profiles of infiltrating T cells in lupus skin and kidney.
Nicotine Exacerbates TAAD Formation Induced by Smooth Muscle-Specific Deletion of the TGF-β Receptor 2.
Bioengineering the Oxygen-Deprived Tumor Microenvironment Within a Three-Dimensional Platform for Studying Tumor-Immune Interactions.
Efficacy and safety of camrelizumab combined with apatinib in advanced triple-negative breast cancer: an open-label phase II trial.
Tryptophan 2,3-Dioxygenase Expression Identified in Human Hepatocellular Carcinoma Cells and in Intratumoral Pericytes of Most Cancers.
Immune risk assessment of residual αGal in xenogeneic decellularized cornea using GTKO mice.
Dataset on the identification of a prognostic radio-immune signature in surgically resected Non Small Cell Lung Cancer.
CD103(+) tumor-infiltrating lymphocytes predict favorable prognosis in patients with esophageal squamous cell carcinoma.
Prognostic impact of CD8 and programmed death-ligand 1 expression in patients with resectable non-small cell lung cancer.
Tracing Oncogene Rearrangements in the Mutational History of Lung Adenocarcinoma.
Cooperation between Constitutive and Inducible Chemokines Enables T Cell Engraftment and Immune Attack in Solid Tumors.
Alterations in PD-L1 Expression Associated with Acquisition of Resistance to ALK Inhibitors in ALK-Rearranged Lung Cancer.
Cellular Composition and Contribution of Tertiary Lymphoid Structures to Tumor Immune Infiltration and Modulation by Radiation Therapy.
Immune Checkpoint Inhibition Overcomes ADCP-Induced Immunosuppression by Macrophages.
Immunoscore in mismatch repair-proficient and -deficient colon cancer.
Prognostic implications of intratumoral CD103+ tumor-infiltrating lymphocytes in pulmonary squamous cell carcinoma.
CD8+ T cells promote proliferation of benign prostatic hyperplasia epithelial cells under low androgen level via modulation of CCL5/STAT5/CCND1 signaling pathway.
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
Clinical Implications of Cytotoxic T Lymphocyte Antigen-4 Expression on Tumor Cells and Tumor-Infiltrating Lymphocytes in Extrahepatic Bile Duct Cancer Patients Undergoing Surgery Plus Adjuvant Chemoradiotherapy.
Expression of programmed cell death ligand 1 (PD-L1) in advanced stage EBV-associated extranodal NK/T cell lymphoma is associated with better prognosis.
Nontuberculous mycobacterial infection with concurrent IgG4-related lymphadenopathy.
Evidence of a pathogenic role for CD8(+) T cells in anti-GABAB receptor limbic encephalitis.
Prognostic Implication of M2 Macrophages Are Determined by the Proportional Balance of Tumor Associated Macrophages and Tumor Infiltrating Lymphocytes in Microsatellite-Unstable Gastric Carcinoma.
Tumor-Infiltrating Immune Cells Are Associated With Prognosis of Gastric Cancer.
CUX1 modulates polarization of tumor-associated macrophages by antagonizing NF-κB signaling.
Low intraprostatic DHT promotes the infiltration of CD8+ T cells in BPH tissues via modulation of CCL5 secretion.
Prognostic implications of tumor-infiltrating FoxP3+ regulatory T cells and CD8+ cytotoxic T cells in microsatellite-unstable gastric cancers.
Expression of CD8 is associated with non-common type morphology and outcome in pediatric anaplastic lymphoma kinase-positive anaplastic large cell lymphoma.
Tumor-infiltrating lymphocytes, tumor characteristics, and recurrence in patients with early breast cancer.
T cell-activation in neuromyelitis optica lesions plays a role in their formation.
Analysis of 142 Northern Chinese patients with peripheral T/NK-Cell lymphomas: subtype distribution, clinicopathologic features, and prognosis.
Lymphoid tissue damage in HIV-1 infection depletes naïve T cells and limits T cell reconstitution after antiretroviral therapy.
Anaplastic large cell lymphoma with paraneoplastic leukocytosis: a clinicopathological analysis of five cases.
Hepatitis C virus induces the expression of CCL17 and CCL22 chemokines that attract regulatory T cells to the site of infection.
CD8⁺ T cells with an intraepithelial phenotype upregulate cytotoxic function upon influenza infection in human lung.
Study of cellular immune response against Hepatitis E virus (HEV).
Spatial differences in the presence of FOXP3+ and GranzymeB+ T cells between the intra- and extravascular compartments in renal allograft vasculopathy.
Primary central nervous system peripheral T-cell lymphoma in a child.
Antagonism of TIM-1 blocks the development of disease in a humanized mouse model of allergic asthma.
Increased T-bet+ cytotoxic effectors and type I interferon-mediated processes in chronic graft-versus-host disease of the oral mucosa.
GMP-17-positive T-lymphocytes in renal tubules predict progression in early stages of IgA nephropathy.
Selective expansion of influenza A virus-specific T cells in symptomatic human carotid artery atherosclerotic plaques.
Effect of melatonin and zinc on the immune response in experimental Toxoplasma retinochoroiditis.
Intrahepatic virus-specific IL-10-producing CD8 T cells prevent liver damage during chronic hepatitis C virus infection.
Dunlap GS, Billi AC, Xing X, Ma F, Maz MP, Tsoi LC, Wasikowski R, Hodgin JB, Gudjonsson JE, Kahlenberg JM, Rao DA
JCI insight 2022 Apr 22;7(8)
JCI insight 2022 Apr 22;7(8)
Nicotine Exacerbates TAAD Formation Induced by Smooth Muscle-Specific Deletion of the TGF-β Receptor 2.
Chun C, Qi X, Wang F, Madrid KB, Saldarriaga LA, Fisch MR, Brantly ML, Upchurch GR Jr, Jiang Z
Journal of immunology research 2021;2021:6880036
Journal of immunology research 2021;2021:6880036
Bioengineering the Oxygen-Deprived Tumor Microenvironment Within a Three-Dimensional Platform for Studying Tumor-Immune Interactions.
Bhattacharya S, Calar K, Evans C, Petrasko M, de la Puente P
Frontiers in bioengineering and biotechnology 2020;8:1040
Frontiers in bioengineering and biotechnology 2020;8:1040
Efficacy and safety of camrelizumab combined with apatinib in advanced triple-negative breast cancer: an open-label phase II trial.
Liu J, Liu Q, Li Y, Li Q, Su F, Yao H, Su S, Wang Q, Jin L, Wang Y, Lau WY, Jiang Z, Song E
Journal for immunotherapy of cancer 2020 May;8(1)
Journal for immunotherapy of cancer 2020 May;8(1)
Tryptophan 2,3-Dioxygenase Expression Identified in Human Hepatocellular Carcinoma Cells and in Intratumoral Pericytes of Most Cancers.
Hoffmann D, Dvorakova T, Stroobant V, Bouzin C, Daumerie A, Solvay M, Klaessens S, Letellier MC, Renauld JC, van Baren N, Lelotte J, Marbaix E, Van den Eynde BJ
Cancer immunology research 2020 Jan;8(1):19-31
Cancer immunology research 2020 Jan;8(1):19-31
Immune risk assessment of residual αGal in xenogeneic decellularized cornea using GTKO mice.
Chen L, Wei L, Shao A, Xu L
Regenerative biomaterials 2020 Aug;7(4):427-434
Regenerative biomaterials 2020 Aug;7(4):427-434
Dataset on the identification of a prognostic radio-immune signature in surgically resected Non Small Cell Lung Cancer.
Mazzaschi G, Milanese G, Pagano P, Madeddu D, Gnetti L, Trentini F, Falco A, Frati C, Lorusso B, Lagrasta C, Minari R, Ampollini L, Silva M, Sverzellati N, Quaini F, Roti G, Tiseo M
Data in brief 2020 Aug;31:105781
Data in brief 2020 Aug;31:105781
CD103(+) tumor-infiltrating lymphocytes predict favorable prognosis in patients with esophageal squamous cell carcinoma.
Chu Y, Liao J, Li J, Wang Y, Yu X, Wang J, Xu X, Xu L, Zheng L, Xu J, Li L
Journal of Cancer 2019;10(21):5234-5243
Journal of Cancer 2019;10(21):5234-5243
Prognostic impact of CD8 and programmed death-ligand 1 expression in patients with resectable non-small cell lung cancer.
Kim SH, Go SI, Song DH, Park SW, Kim HR, Jang I, Kim JD, Lee JS, Lee GW
British journal of cancer 2019 Mar;120(5):547-554
British journal of cancer 2019 Mar;120(5):547-554
Tracing Oncogene Rearrangements in the Mutational History of Lung Adenocarcinoma.
Lee JJ, Park S, Park H, Kim S, Lee J, Lee J, Youk J, Yi K, An Y, Park IK, Kang CH, Chung DH, Kim TM, Jeon YK, Hong D, Park PJ, Ju YS, Kim YT
Cell 2019 Jun 13;177(7):1842-1857.e21
Cell 2019 Jun 13;177(7):1842-1857.e21
Cooperation between Constitutive and Inducible Chemokines Enables T Cell Engraftment and Immune Attack in Solid Tumors.
Dangaj D, Bruand M, Grimm AJ, Ronet C, Barras D, Duttagupta PA, Lanitis E, Duraiswamy J, Tanyi JL, Benencia F, Conejo-Garcia J, Ramay HR, Montone KT, Powell DJ Jr, Gimotty PA, Facciabene A, Jackson DG, Weber JS, Rodig SJ, Hodi SF, Kandalaft LE, Irving M, Zhang L, Foukas P, Rusakiewicz S, Delorenzi M, Coukos G
Cancer cell 2019 Jun 10;35(6):885-900.e10
Cancer cell 2019 Jun 10;35(6):885-900.e10
Alterations in PD-L1 Expression Associated with Acquisition of Resistance to ALK Inhibitors in ALK-Rearranged Lung Cancer.
Kim SJ, Kim S, Kim DW, Kim M, Keam B, Kim TM, Lee Y, Koh J, Jeon YK, Heo DS
Cancer research and treatment 2019 Jul;51(3):1231-1240
Cancer research and treatment 2019 Jul;51(3):1231-1240
Cellular Composition and Contribution of Tertiary Lymphoid Structures to Tumor Immune Infiltration and Modulation by Radiation Therapy.
Boivin G, Kalambaden P, Faget J, Rusakiewicz S, Montay-Gruel P, Meylan E, Bourhis J, Lesec G, Vozenin MC
Frontiers in oncology 2018;8:256
Frontiers in oncology 2018;8:256
Immune Checkpoint Inhibition Overcomes ADCP-Induced Immunosuppression by Macrophages.
Su S, Zhao J, Xing Y, Zhang X, Liu J, Ouyang Q, Chen J, Su F, Liu Q, Song E
Cell 2018 Oct 4;175(2):442-457.e23
Cell 2018 Oct 4;175(2):442-457.e23
Immunoscore in mismatch repair-proficient and -deficient colon cancer.
Wirta EV, Seppälä T, Friman M, Väyrynen J, Ahtiainen M, Kautiainen H, Kuopio T, Kellokumpu I, Mecklin JP, Böhm J
The journal of pathology. Clinical research 2017 Jul;3(3):203-213
The journal of pathology. Clinical research 2017 Jul;3(3):203-213
Prognostic implications of intratumoral CD103+ tumor-infiltrating lymphocytes in pulmonary squamous cell carcinoma.
Koh J, Kim S, Kim MY, Go H, Jeon YK, Chung DH
Oncotarget 2017 Feb 21;8(8):13762-13769
Oncotarget 2017 Feb 21;8(8):13762-13769
CD8+ T cells promote proliferation of benign prostatic hyperplasia epithelial cells under low androgen level via modulation of CCL5/STAT5/CCND1 signaling pathway.
Yang Y, Hu S, Liu J, Cui Y, Fan Y, Lv T, Liu L, Li J, He Q, Han W, Yu W, Sun Y, Jin J
Scientific reports 2017 Feb 20;7:42893
Scientific reports 2017 Feb 20;7:42893
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
Clinical Implications of Cytotoxic T Lymphocyte Antigen-4 Expression on Tumor Cells and Tumor-Infiltrating Lymphocytes in Extrahepatic Bile Duct Cancer Patients Undergoing Surgery Plus Adjuvant Chemoradiotherapy.
Lim YJ, Koh J, Kim K, Chie EK, Kim S, Lee KB, Jang JY, Kim SW, Oh DY, Bang YJ
Targeted oncology 2017 Apr;12(2):211-218
Targeted oncology 2017 Apr;12(2):211-218
Expression of programmed cell death ligand 1 (PD-L1) in advanced stage EBV-associated extranodal NK/T cell lymphoma is associated with better prognosis.
Kim WY, Jung HY, Nam SJ, Kim TM, Heo DS, Kim CW, Jeon YK
Virchows Archiv : an international journal of pathology 2016 Nov;469(5):581-590
Virchows Archiv : an international journal of pathology 2016 Nov;469(5):581-590
Nontuberculous mycobacterial infection with concurrent IgG4-related lymphadenopathy.
Liu TT, Weng SW, Wang MC, Huang WT
APMIS : acta pathologica, microbiologica, et immunologica Scandinavica 2016 Mar;124(3):216-20
APMIS : acta pathologica, microbiologica, et immunologica Scandinavica 2016 Mar;124(3):216-20
Evidence of a pathogenic role for CD8(+) T cells in anti-GABAB receptor limbic encephalitis.
Golombeck KS, Bönte K, Mönig C, van Loo KM, Hartwig M, Schwindt W, Widman G, Lindenau M, Becker AJ, Glatzel M, Elger CE, Wiendl H, Meuth SG, Lohmann H, Gross CC, Melzer N
Neurology(R) neuroimmunology & neuroinflammation 2016 Jun;3(3):e232
Neurology(R) neuroimmunology & neuroinflammation 2016 Jun;3(3):e232
Prognostic Implication of M2 Macrophages Are Determined by the Proportional Balance of Tumor Associated Macrophages and Tumor Infiltrating Lymphocytes in Microsatellite-Unstable Gastric Carcinoma.
Kim KJ, Wen XY, Yang HK, Kim WH, Kang GH
PloS one 2015;10(12):e0144192
PloS one 2015;10(12):e0144192
Tumor-Infiltrating Immune Cells Are Associated With Prognosis of Gastric Cancer.
Liu K, Yang K, Wu B, Chen H, Chen X, Chen X, Jiang L, Ye F, He D, Lu Z, Xue L, Zhang W, Li Q, Zhou Z, Mo X, Hu J
Medicine 2015 Sep;94(39):e1631
Medicine 2015 Sep;94(39):e1631
CUX1 modulates polarization of tumor-associated macrophages by antagonizing NF-κB signaling.
Kühnemuth B, Mühlberg L, Schipper M, Griesmann H, Neesse A, Milosevic N, Wissniowski T, Buchholz M, Gress TM, Michl P
Oncogene 2015 Jan 8;34(2):177-87
Oncogene 2015 Jan 8;34(2):177-87
Low intraprostatic DHT promotes the infiltration of CD8+ T cells in BPH tissues via modulation of CCL5 secretion.
Fan Y, Hu S, Liu J, Xiao F, Li X, Yu W, Cui Y, Sun M, Lv T, He Q, Jin J
Mediators of inflammation 2014;2014:397815
Mediators of inflammation 2014;2014:397815
Prognostic implications of tumor-infiltrating FoxP3+ regulatory T cells and CD8+ cytotoxic T cells in microsatellite-unstable gastric cancers.
Kim KJ, Lee KS, Cho HJ, Kim YH, Yang HK, Kim WH, Kang GH
Human pathology 2014 Feb;45(2):285-93
Human pathology 2014 Feb;45(2):285-93
Expression of CD8 is associated with non-common type morphology and outcome in pediatric anaplastic lymphoma kinase-positive anaplastic large cell lymphoma.
Abramov D, Oschlies I, Zimmermann M, Konovalov D, Damm-Welk C, Wössmann W, Klapper W
Haematologica 2013 Oct;98(10):1547-53
Haematologica 2013 Oct;98(10):1547-53
Tumor-infiltrating lymphocytes, tumor characteristics, and recurrence in patients with early breast cancer.
Kim ST, Jeong H, Woo OH, Seo JH, Kim A, Lee ES, Shin SW, Kim YH, Kim JS, Park KH
American journal of clinical oncology 2013 Jun;36(3):224-31
American journal of clinical oncology 2013 Jun;36(3):224-31
T cell-activation in neuromyelitis optica lesions plays a role in their formation.
Pohl M, Kawakami N, Kitic M, Bauer J, Martins R, Fischer MT, Machado-Santos J, Mader S, Ellwart JW, Misu T, Fujihara K, Wekerle H, Reindl M, Lassmann H, Bradl M
Acta neuropathologica communications 2013 Dec 24;1:85
Acta neuropathologica communications 2013 Dec 24;1:85
Analysis of 142 Northern Chinese patients with peripheral T/NK-Cell lymphomas: subtype distribution, clinicopathologic features, and prognosis.
Ren YL, Nong L, Zhang S, Zhao J, Zhang XM, Li T
American journal of clinical pathology 2012 Sep;138(3):435-47
American journal of clinical pathology 2012 Sep;138(3):435-47
Lymphoid tissue damage in HIV-1 infection depletes naïve T cells and limits T cell reconstitution after antiretroviral therapy.
Zeng M, Southern PJ, Reilly CS, Beilman GJ, Chipman JG, Schacker TW, Haase AT
PLoS pathogens 2012 Jan;8(1):e1002437
PLoS pathogens 2012 Jan;8(1):e1002437
Anaplastic large cell lymphoma with paraneoplastic leukocytosis: a clinicopathological analysis of five cases.
Chang IW, Chen HK, Ma MC, Huang WT
APMIS : acta pathologica, microbiologica, et immunologica Scandinavica 2011 Nov;119(11):794-801
APMIS : acta pathologica, microbiologica, et immunologica Scandinavica 2011 Nov;119(11):794-801
Hepatitis C virus induces the expression of CCL17 and CCL22 chemokines that attract regulatory T cells to the site of infection.
Riezu-Boj JI, Larrea E, Aldabe R, Guembe L, Casares N, Galeano E, Echeverria I, Sarobe P, Herrero I, Sangro B, Prieto J, Lasarte JJ
Journal of hepatology 2011 Mar;54(3):422-31
Journal of hepatology 2011 Mar;54(3):422-31
CD8⁺ T cells with an intraepithelial phenotype upregulate cytotoxic function upon influenza infection in human lung.
Piet B, de Bree GJ, Smids-Dierdorp BS, van der Loos CM, Remmerswaal EB, von der Thüsen JH, van Haarst JM, Eerenberg JP, ten Brinke A, van der Bij W, Timens W, van Lier RA, Jonkers RE
The Journal of clinical investigation 2011 Jun;121(6):2254-63
The Journal of clinical investigation 2011 Jun;121(6):2254-63
Study of cellular immune response against Hepatitis E virus (HEV).
Prabhu SB, Gupta P, Durgapal H, Rath S, Gupta SD, Acharya SK, Panda SK
Journal of viral hepatitis 2011 Aug;18(8):587-94
Journal of viral hepatitis 2011 Aug;18(8):587-94
Spatial differences in the presence of FOXP3+ and GranzymeB+ T cells between the intra- and extravascular compartments in renal allograft vasculopathy.
de Boer OJ, Teeling P, Jansen M, Ploegmakers H, van der Loos CM, Kummer JA, Florquin S, van der Wal AC
PloS one 2011 Apr 6;6(4):e18656
PloS one 2011 Apr 6;6(4):e18656
Primary central nervous system peripheral T-cell lymphoma in a child.
Gualco G, Wludarski S, Hayashi-Silva L, Medeiros Filho P, Veras G, Bacchi CE
Fetal and pediatric pathology 2010;29(4):224-30
Fetal and pediatric pathology 2010;29(4):224-30
Antagonism of TIM-1 blocks the development of disease in a humanized mouse model of allergic asthma.
Sonar SS, Hsu YM, Conrad ML, Majeau GR, Kilic A, Garber E, Gao Y, Nwankwo C, Willer G, Dudda JC, Kim H, Bailly V, Pagenstecher A, Rennert PD, Renz H
The Journal of clinical investigation 2010 Aug;120(8):2767-81
The Journal of clinical investigation 2010 Aug;120(8):2767-81
Increased T-bet+ cytotoxic effectors and type I interferon-mediated processes in chronic graft-versus-host disease of the oral mucosa.
Imanguli MM, Swaim WD, League SC, Gress RE, Pavletic SZ, Hakim FT
Blood 2009 Apr 9;113(15):3620-30
Blood 2009 Apr 9;113(15):3620-30
GMP-17-positive T-lymphocytes in renal tubules predict progression in early stages of IgA nephropathy.
van Es LA, de Heer E, Vleming LJ, van der Wal A, Mallat M, Bajema I, Bruijn JA, de Fijter JW
Kidney international 2008 Jun;73(12):1426-33
Kidney international 2008 Jun;73(12):1426-33
Selective expansion of influenza A virus-specific T cells in symptomatic human carotid artery atherosclerotic plaques.
Keller TT, van der Meer JJ, Teeling P, van der Sluijs K, Idu MM, Rimmelzwaan GF, Levi M, van der Wal AC, de Boer OJ
Stroke 2008 Jan;39(1):174-9
Stroke 2008 Jan;39(1):174-9
Effect of melatonin and zinc on the immune response in experimental Toxoplasma retinochoroiditis.
Avunduk AM, Avunduk MC, Baltaci AK, Moğulkoç R
Ophthalmologica. Journal international d'ophtalmologie. International journal of ophthalmology. Zeitschrift fur Augenheilkunde 2007;221(6):421-5
Ophthalmologica. Journal international d'ophtalmologie. International journal of ophthalmology. Zeitschrift fur Augenheilkunde 2007;221(6):421-5
Intrahepatic virus-specific IL-10-producing CD8 T cells prevent liver damage during chronic hepatitis C virus infection.
Abel M, Sène D, Pol S, Bourlière M, Poynard T, Charlotte F, Cacoub P, Caillat-Zucman S
Hepatology (Baltimore, Md.) 2006 Dec;44(6):1607-16
Hepatology (Baltimore, Md.) 2006 Dec;44(6):1607-16
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Formalin-fixed, paraffin-embedded human tonsil stained with CD8 and AEC chromogen.
Supportive validation
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- Invitrogen Antibodies (provider)
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- Flow cytometry analysis of CD8 in PBMC cells (green) compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 µL of cell solution was added to each tube at a dilution of 2x10^7 cells/mL, followed by the addition of 50 µL of isotype control and primary antibody (Product # MA5-14548) at a dilution of 1:2. Cells were incubated for 30 min at 4ºC and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated secondary antibody for 30 min at 4ºC in the dark. FACS analysis was performed using 400 µL of cell buffer.
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- Flow cytometry analysis of CD8 in Jurkat cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a CD8 monoclonal antibody (Product # MA5-14548) at a dilution of 1:2 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.
Supportive validation
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- Figure 2 CD4 + and CD8 + T cells accumulate in the CSF in GABA B receptor antibody-associated LE Multiparameter flow cytometry analysis of absolute numbers of CD4 + T cells (A) and CD8 + T cells (C), activated HLADR + CD4 + T cells (B), and HLADR + CD8 + T cells (D) within PB and CSF in 4 patients with GABA B receptor antibody-associated LE and controls. Data are given as whisker plots. Levels of significance are indicated by n.s. (not significant) for all p > 0.05, * for all p < 0.05, and ** for all p < 0.01. For details, refer to the main text and table 1 . GABA B = gamma-aminobutyric acid B; LE = limbic encephalitis; PB = peripheral blood; Pt. = patient.
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- Figure 3 A pathogenic role for CD8 + T cells in addition to plasma cell-derived autoantibodies in GABA B receptor antibody-associated limbic encephalitis Histologic specimen of one patient with GABA B receptor antibody-associated limbic encephalitis following selective amygdalohippocampectomy revealed perivascular infiltrates of CD138 + plasma cells (A) and CD3 + T cells (B). CD8 + T cells (C) were predominantly detected within the brain parenchyma in close contact to neurons but not in perivascular regions, suggesting that perivascular CD3 + T cells are mainly CD4 + T cells (scale bar represents 50 mum in A-C; insets show magnification of 2 representative neurons in close contact with CD8 + T cells). Representative confocal triple staining (D) for DAPI, CD8 (red), and granzyme B (green) revealed expression of granzyme B (white arrow) in parenchymal CD8 + T cells (scale bar represents 10 mum in D). DAPI = 4'6-diamidino-2-phenylindole*2HCl; GABA B = gamma-aminobutyric acid B.
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- Figure 4 HE and immunohistochemical staining of infiltrating cells in decellularized corneal matrix after 4 weeks of implantation in the WT mice and GTKO mice. Figures of ( A ) HE staining, ( C ) CD68 + , ( E ) CD3 + , ( F ) CD4 + , and ( G ) CD8 + were obtained from WT mice; ( B ) HE staining, ( D ) CD68 + , ( H ) CD3 + , ( I ) CD4 + , and ( J ) CD8 + were images from GTKO mice. The black arrows showed the positive staining cells. DCM: decellularized corneal matrix.
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- FIGURE 7 Low-oxygen within 3D-O matrices reduces lymphocyte infiltration. (A) tSNE plot of pre-processed, single, live cells, down-sampled to 10.000 events and concatenated showing major immune clusters (CD3-FITC+, CD4-PE-Cy5+, CD8-APC-Cy7+, CD19-APC+ and CD45-BV510+). (B) Quantification of the number of infiltrated lymphocytes into 3D-O physiological and 3D-O tumorous matrices embedded with either MDA-MB-231 cells or MCF-7 cells by manual gating. Infiltration data shown represents PBMCs obtained from 3 individual healthy subjects and the average of infiltrated CD3+ T cells, CD3+CD8+ T cells, CD3+CD4+ T cells, and CD19+ B cells (data normalized to counting beads x10 3 ), * p < 0.05, data were analyzed using unpaired two-tailed Student''s t -test. (C) Representative confocal microscopy images of rotated and top views of BCa (green) cells and CD8+ (red) cells in 3D-O physiological and 3D-O tumorous matrices on day 7, co-localization is seen in yellow. Scale bar = 200 mum.
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- FIGURE 8 Sensitization of BCa cells to cytotoxic CD8+ T cells within 3D-O matrices. (A) CD8+ infiltration into 3D-O physiological and 3D-O tumorous matrices seeded with MDA-MB-231 and MCF-7 cells on day 7 after treatment with PX-478 at 5 muM concentration (i) and Durvalumab at 5 muM concentration (ii) for the first 4 days. (B) Representative IHC results representing CD8+ infiltration into 3D-physiological and 3D-tumorous scaffolds seeded with MDA-MB-231 and MCF-7 cells on day 7 after treatment with PX-478 and Durvalumab, Scale bar = 300 mum. * p < 0.05, (n.s.) not significant, data were analyzed using unpaired two-tailed Student's t -test.
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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 2 Cellular source of CD103 + cells in ESCC tissues. (A) Double immunofluorescence staining shows CD103 (green), CD3 (red), CD11c (red), CD4 (red), and CD8 (red) expression and co-localization of double-positive cells (yellow) in ESCC tissue. (B) The percentages of CD11c + CD103 + cells identified as CD11c and CD103 double-positive cells and calculated in total numbers of CD103 + cells in the ANT and IT regions. Scale bar = 25 mum. (C, D) The percentages of CD3 + CD103 + cells identified as double-positive cells and calculated in total numbers of CD103 + or CD3 + cells in the ANT and IT regions ( n = 7). (E, F) The percentages of CD4 + CD103 + and CD8 + CD103 + cells identified as double-positive cells and compared to the total numbers of CD103 + cells in the ANT (E) and IT regions (F), respectively ( n = 10). Results are the means +- SEM (bars); ** P < 0.01; *** P < 0.001.