- 13-1529-82 - Invitrogen Antibodies CTLA4 antibody

Submitted references Cancer stem-like cells evade CD8(+)CD103(+) tumor-resident memory T (T(RM)) lymphocytes by initiating an epithelial-to-mesenchymal transition program in a human lung tumor model.

Corgnac S, Damei I, Gros G, Caidi A, Terry S, Chouaib S, Deloger M, Mami-Chouaib F
Journal for immunotherapy of cancer 2022 Apr;10(4)

Engineering advanced logic and distributed computing in human CAR immune cells.

Cho JH, Okuma A, Sofjan K, Lee S, Collins JJ, Wong WW
Nature communications 2021 Feb 4;12(1):792

Methylome-based cell-of-origin modeling (Methyl-COOM) identifies aberrant expression of immune regulatory molecules in CLL.

Wierzbinska JA, Toth R, Ishaque N, Rippe K, Mallm JP, Klett LC, Mertens D, Zenz T, Hielscher T, Seifert M, Küppers R, Assenov Y, Lutsik P, Stilgenbauer S, Roessner PM, Seiffert M, Byrd J, Oakes CC, Plass C, Lipka DB
Genome medicine 2020 Mar 18;12(1):29

Accumulation of TNFR2-expressing regulatory T cells in malignant pleural effusion of lung cancer patients is associated with poor prognosis.

Ye LL, Peng WB, Niu YR, Xiang X, Wei XS, Wang ZH, Wang X, Zhang SY, Chen X, Zhou Q
Annals of translational medicine 2020 Dec;8(24):1647

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

Low Expression of miR-424-3p is Highly Correlated with Clinical Failure in Prostate Cancer.

Richardsen E, Andersen S, Al-Saad S, Rakaee M, Nordby Y, Pedersen MI, Ness N, Ingebriktsen LM, Fassina A, Taskén KA, Mills IG, Donnem T, Bremnes RM, Busund LT
Scientific reports 2019 Jul 23;9(1):10662

CD226 opposes TIGIT to disrupt Tregs in melanoma.

Fourcade J, Sun Z, Chauvin JM, Ka M, Davar D, Pagliano O, Wang H, Saada S, Menna C, Amin R, Sander C, Kirkwood JM, Korman AJ, Zarour HM
JCI insight 2018 Jul 26;3(14)

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

Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.

Miles B, Miller SM, Folkvord JM, Kimball A, Chamanian M, Meditz AL, Arends T, McCarter MD, Levy DN, Rakasz EG, Skinner PJ, Connick E
Nature communications 2015 Oct 20;6:8608

Investigation of soluble and transmembrane CTLA-4 isoforms in serum and microvesicles.

Esposito L, Hunter KM, Clark J, Rainbow DB, Stevens H, Denesha J, Duley S, Dawson S, Coleman G, Nutland S, Bell GL, Moran C, Pekalski M, Todd JA, Wicker LS
Journal of immunology (Baltimore, Md. : 1950) 2014 Jul 15;193(2):889-900

Programmed death-1+ T cells and regulatory T cells are enriched in tumor-involved lymph nodes and associated with aggressive features in papillary thyroid cancer.

French JD, Kotnis GR, Said S, Raeburn CD, McIntyre RC Jr, Klopper JP, Haugen BR
The Journal of clinical endocrinology and metabolism 2012 Jun;97(6):E934-43

Soluble CTLA-4 in autoimmune thyroid diseases: relationship with clinical status and possible role in the immune response dysregulation.

Saverino D, Brizzolara R, Simone R, Chiappori A, Milintenda-Floriani F, Pesce G, Bagnasco M
Clinical immunology (Orlando, Fla.) 2007 May;123(2):190-8

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: PHA-stimulated human peripheral blood cells were intracellularly stained with 0.03 µg of Mouse IgG2a K Isotype Control Biotin (Product # 13-4724-85) (open histogram) or 0.03 µg of Anti-Human CD152 (CTLA-4) Biotin (filled histogram) followed by Streptavidin PE (Product # 12-4317-87) using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Total viable cells were used for analysis.
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 T FR exhibit an enhanced regulatory phenotype in ex vivo HIV infection. Tonsil cells were mock-, X4-, or R5-spinoculated and cultured under experimental conditions as indicated. T FR were then analysed for expression of regulatory receptors and cytokine production by intracellular cytokine staining. ( a ) Percentage of total (surface and intracellular) T FR CTLA-4 expression ( n =15). ( b ) Percentage of surface T FR LAG-3 expression ( n =8). ( c ) Production of IL-10 by T FR ( n =7). ( d ) Production of TGF-beta-1 (measured as LAP) by T FR ( n =5). The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Friedman ( a , b ) or Mann-Whitney tests ( c , d ) and significance is denoted by asterisks where * P
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Resident memory T (T RM ) and non-T RM phenotypic profiles and susceptibility of cancer stem cell (CSC) to cytotoxic T lymphocyte (CTL)-mediated killing. (A) Flow cytometry analyses of CD8, LFA-1, CD103, CD49a, CD69, PD-1, CTLA-4, TGFBR2 and VEGFR2 on CD8 + CD103 + T RM (Heu171) and CD8 + CD103 - non-T RM (H32-22) clones. Mean immunofluorescence intensity (MFI) are in parentheses. (B) Cytotoxic activity of the T RM clone (Heu171) towards autologous IGR-Heu, Heu-CSC, CSC-1 and CSC-2 target cells. Percent of specific lysis are shown at indicated effector to target (E:T) ratios. (C) Quantification of transforming growth factor (TGF)-beta in conditioned media from IGR-Heu, Heu-CSC, CSC-1 and CSC-2 by multi-analyte flow assay. Ratios of active/total TGF-beta normalized to IGR-Heu are included. Results are presented as mean+-SEM of duplicates. (D) Cytotoxicity of the non-T RM clone (H32-22) towards autologous IGR-Heu, Heu-CSC, CSC-1 and CSC-2 target cells. (E) Inhibition of T RM -cell-mediated killing. CSC-1 cells are preincubated in the presence of isotype control, anti-major histocompatibility complex class I (MHC-I) or anti-intercellular adhesion molecule 1 (ICAM-1) neutralizing monoclonal antibody (mAb) and then CTL were added at 5:1 E:T ratio. Symbols represent replicates and horizontal bars represent means+-SEM (n=3). P value was determined by two-way analysis of variance test. *p
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Effects of TNF on the suppressive function of TNFR2 + Tregs. Purified CD4 + T cells were cultured in medium alone, TNF (50 ng/mL), TNF combined with anti-TNFR2 mAbs (10 ug/mL), or isotype IgG, as indicated, for 72 hours. The representative FACS analysis of (A) CTLA-4 + cells and (B) PD-L1 + cells in TNFR2 + Tregs as indicated, gated on live CD4 + Foxp3 + cells. Summary of the proportions of (C) CTLA-4 + cells (n=3) and (D) PD-L1 + cells (n=3) in Tregs within each condition. Data are expressed as means +- SEM. *, P
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 T FR expansion in lymphoid tissues during chronic SIV infection. ( a ) Disaggregated lymph node and spleen cells from SIV uninfected ( n =9) or chronically SIV-infected rhesus macaques ( n =11) were analysed by flow cytometry. Representative examples of flow cytometry gating are shown. Of viable CD3 + CD8 - cells, follicular subsets were defined as CXCR5 + cells (F) and germinal centre subsets were defined as CXCR5 hi PD-1 hi cells (GC). Of these subsets, regulatory cells were defined as CD25 hi CD127 - . T FR (CXCR5 + CD25 hi CD127 - ) were Foxp3 + , whereas T FH (CXCR5 + CD25 lo/- ) were Foxp3 - . ( b ) The percentages of each rhesus macaque regulatory subset, as analysed in a are shown. ( c ) The ratios of each regulatory cell population to its non-regulatory cell counterpart are shown. ( d ) The percentage of total CTLA-4 expression is shown in SIV-uninfected ( n =9) and chronically SIV-infected ( n =8) rhesus macaques. The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Mann-Whitney (Wilcoxon) tests to compare unpaired, nonparametric values and significance is denoted by asterisks where * P
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Panel of CTLA-4 immunohistochemical staining. ( A ) Strong nuclear staining expression of CTLA-4, Gleason grade group 1 (3 + 3); ( B ) Moderate staining expression, Gleason grade group 4 (4 + 4); ( C ) Weak staining expression, Gleason grade group 5 (4 + 5). (magnification 20x).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 CD103 + cells co-express CTLA-4, granzyme B (GB), and PD-1 in ESCC. (A-C) Double immunofluorescence staining shows CD103, CTLA-4 (A), granzyme B (GB) (B), and PD-1 (C) expression and co-localization of double-positive cells in ESCC tissue. DAPI (blue) was used as a counterstain. Scale bar = 25 mum. The percentages were identified as double-positive cells compared to the total numbers of CD103 + cells in the ANT and T regions, respectively ( n = 7). Results are the means +- SEM (bars).
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 Flow cytometry analysis of T cell-/lymphocyte-specific markers on normal and malignant B cells from CLL patients. a Summary scheme representing functional implications of CLL-specific candidate genes selected for flow cytometric analysis. b Flow cytometric analysis of expression of CTLA-4, TIGIT, CD276, LILRB4, and CD2 on peripheral blood B cells of CLL patients. The expression was determined for non-malignant B cells (""Normal""; CD19 + CD5 - B cells, represented in green) and neoplastic B cells (""CLL"", CD19 + CD5 + B cells, represented in orange) detected in the same samples. ""Co,"" no antibody staining control; ""Ab,"" staining with the antibody of interest as indicated. c Normalized median fluorescence intensities (target MFI - MFI of negative control [Co]; nMFI). d Delta normalized median fluorescence intensities between CLL cells and normal B cells (DeltanMFI (CLL-normal)) for each patient tested
Conjugate: Biotin

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 The intracellular AND logic with different signaling domains. a Diagram of intracellular AND logic. b Primary human CD8+ T cells were transduced with FOS zipCAR-containing CD3zeta domain and RR zipCAR-containing CD28 domain. Cytotoxicity against Her2- and Axl-expressing Nalm6 was measured 24 h after adding alpha-Her2-SYN9 and/or alpha-Axl-EE zipFvs. The heatmap indicates cytotoxicity at varying zipFv concentrations ( n = 3, data are represented as mean). c Cytotoxicity of CD8+ T cells transduced with FOS zipCAR-containing CD3zeta domain and RR zipCAR-containing 4-1BB domain. The heatmap indicates cytotoxicity at varying zipFv concentrations ( n = 3, data are represented as mean). d (Left) Isolated Treg cells were transduced with two zipCAR constructs: SYN6-CD3zeta-P2A-FOXP3 and SYN1-CD28-P2A-puro. After puromycin selection (2 mug/mL), Treg cells were co-cultured with Her2- and Axl-expressing Nalm6 target cells (Right) The heatmap shows surface CTLA-4 expression detected after 48 h by flow cytometry at varying zipFv concentrations (alpha-Axl-SYN5 and alpha-Her2-SYN2) ( n = 3, data are represented as mean).
Conjugate: Biotin

13-1529-82

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