Cd8 clone yts 169

Manufactured by BioXCell

CD8 (clone YTS 169.4) is a laboratory reagent used in cell biology research. It is an anti-CD8 monoclonal antibody that recognizes the CD8 cell surface antigen, which is expressed on certain T cells and natural killer cells. The antibody can be used for the identification and isolation of CD8-positive cells in various experimental systems.

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Lab products found in correlation

Cd4 clone gk1, by BioXCell (3 mentions) Ifnγ clone xmg1.2, by BioXCell (2 mentions) Cd4 t cells clone gk1.5, by BioXCell (1 mentions) Nod scid il2rgammanull nsg mice, by Jackson ImmunoResearch (1 mentions) Matrigel, by Bio-Techne (1 mentions) Wild type c57bl 6 mice, by Taconic Biosciences (1 mentions) Clone ltf 2, by BioXCell (1 mentions) Clone ebr2a, by Thermo Fisher Scientific (1 mentions) Simvastatin, by Avantor (1 mentions) Propylene glycol, by Merck Group (1 mentions) Polyethylene glycol 400, by Merck Group (1 mentions) Clone rmp1 14, by BioXCell (1 mentions) Rat igg2b, by BioXCell (1 mentions) Mouse igg2a clone c1, by BioXCell (1 mentions) Nk1.1 clone pk136, by BioXCell (1 mentions) Pd l1, by BioLegend (1 mentions) Recombinant human interferon gamma ifnγ, by BioLegend (1 mentions) Cd137 41bb, by BioLegend (1 mentions) H 2kb h 2db, by BioLegend (1 mentions) Cd11b, by BioLegend (1 mentions)

8 protocols using cd8 clone yts 169

Anti-PD-1 and CD4/CD8 Depletion

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PD-1 mAb (clone RMP1-14, BioXCell) was administered i.p. at 250 mg/kg daily on days 3–5 and 13–15. CD4 (clone GK1.5, BioXCell) and CD8 (clone YTS 169.4, BioXCell) depleting antibodies were administered i.p. at 200 mg/kg every other day starting 4 days prior to tumor implantation.

Antonios J.P., Soto H., Everson R.G., Orpilla J., Moughon D., Shin N., Sedighim S., Yong W.H., Li G., Cloughesy T.F., Liau L.M, & Prins R.M. (2016). PD-1 blockade enhances the vaccination-induced immune response in glioma. JCI Insight, 1(10), e87059.

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Subcutaneous Tumor Implantation in Mice

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All animals were housed in a barrier facility in air-filtered cages. As indicated for selected experiments 1 × 10⁶ Lewis Lung Carcinoma cells (ATCC®) in the logarithmic phase of growth were injected subcutaneously into the flank. For specific experiments, mice were depleted of NK cells (clone PK136), CD8⁺ (clone YTS169.4) and/or CD4⁺ T cells (clone GK1.5)(BioXcell). C57BL/6 mice expressing cherry tomato-red on an NK1.1 promoter was previously described.⁵⁶ (link) All animal procedures were approved by the Animal Studies Committee at Washington University School of Medicine, St. Louis, Missouri, USA and University of Tübingen and carried out according to the guidelines of the German Law for the Protection of Animals.

Arefanian S., Schäll D., Chang S., Ghasemi R., Higashikubo R., Zheleznyak A., Guo Y., Yu J., Asgharian H., Li W., Gelman A.E., Kreisel D., French A.R., Zaher H., Plougastel-Douglas B., Maggi L., Yokoyama W., Beer-Hammer S, & Krupnick A.S. (2016). Deficiency of the adaptor protein SLy1 results in a natural killer cell ribosomopathy affecting tumor clearance. Oncoimmunology, 5(12), e1238543.

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Syngeneic and Xenograft Tumor Models

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Wild-type C57BL/6 mice were purchased from Taconic and NOD-scid IL2Rgamma^null (NSG) mice were purchased from Jackson Laboratories. Mice were housed in a pathogen-free environment and all experiments were performed under an Animal care and Use Committee approved protocol. Syngeneic murine or xenograft human tumors were established by subcutaneous flank injection of tumor cells in Matrigel (Trevigen, 30% by volume). Mice were assessed for tumor growth three times weekly and tumor volume was calculated as: (length² x width)/2. In some experiments, antibody-based depletion was performed via intraperitoneal (IP) injection of a CD8 (clone YTS 169.4, BioXCell), IFNγ (clone XMG1.2) or TNFα (clone XT3.11) mAb, 200 μg each twice weekly starting 13 days following tumor implantation. Murine blood chemistries were performed by the National Institutes of Health Department of Laboratory Medicine.

Robbins Y., Greene S., Friedman J., Clavijo P.E., Van Waes C., Fabian K.P., Padget M.R., Abdul Sater H., Lee J.H., Soon-Shiong P., Gulley J., Schlom J., Hodge J.W, & Allen C.T. (2020). Tumor control via targeting PD-L1 with chimeric antigen receptor modified NK cells. eLife, 9, e54854.

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Monocyte, Macrophage, and T Cell Depletion

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To deplete monocytes, wildtype C57BL/6 mice received I.V. injection of 200 μL of clodronate liposomes (or PBS control liposomes) 15–18 hours prior to infection [35 (link)]. Anti-CSF1R antibody (2 mg/mL, clone AFS98, eBioscience) was used to deplete bladder-resident macrophages. Animals received two I.V. injections, on consecutive days, of anti-CSF1R antibody or isotype control (clone eBR2a, eBioscience). We administered 400 μg/mouse on day 1 and 200 μg/mouse on day 2, to decrease the impact on circulating monocytes. To deplete T cells, 100 μg of CD4 (clone GK1.5, Bio X Cell) and 100 μg of CD8 (clone YTS 169.4, Bio X Cell) per mouse were injected together intraperitoneally 24 hours prior to primary infection. 200 μg of isotype control (clone LTF-2, Bio X Cell) per mouse was injected intraperitoneally. The depletion was repeated 5 days post-infection, and once a week to maintain the depletion until challenge infection.

Mora-Bau G., Platt A.M., van Rooijen N., Randolph G.J., Albert M.L, & Ingersoll M.A. (2015). Macrophages Subvert Adaptive Immunity to Urinary Tract Infection. PLoS Pathogens, 11(7), e1005044.

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Statin-mediated Immunomodulation in Mice

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Antibodies for in vivo mouse treatments specific for PD-1 (clone RMP1-14) and CD8 (clone YTS 169.4) were from BioXCell. Fluorescent-conjugated flow cytometry antibodies for mouse tumor experiments were obtained from BD Biosciences, Biolegend, Miltenyi or Abcam (see online supplemental methods table S1). Statin drugs (simvastatin, lovastatin, fluvastatin, pitavastatin, atorvastatin, rosuvastatin, pravastatin, table 1), were obtained from VWR International. For in vitro experiments, stock solutions were made with sterile DMSO and frozen at −20°C until use. For in vivo administration, statins were prepared as an oral suspension with 6% polyethylene glycol 400, 1% propylene glycol, and 0.1% Tween (all from Sigma) in water, as previously described.31 (link) Aliquots of statin suspension were frozen at −20°C for up to 2 weeks prior to administration to mice by oral gavage.

Kansal V., Burnham A.J., Kinney B.L., Saba N.F., Paulos C., Lesinski G.B., Buchwald Z.S, & Schmitt N.C. (2023). Statin drugs enhance responses to immune checkpoint blockade in head and neck cancer models. Journal for Immunotherapy of Cancer, 11(1), e005940.

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Depletion of Immune Cells in Teratoma Assay

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Mice were bled (as described in the humanized mice section) prior to the first injection of depleting antibodies. Mice were injected intraperitoneally with 250µg of one of the following depleting antibodies: mouse IgG2a (clone C1.18.4; BioXCell), rat IgG2b (BioXCell), CD8 (clone YTS169.4; BioXCell), CD4 (clone GK1.5; BioXCell), CD20 (clone SA271G2; BioLegend), or NK1.1 (clone PK136; BioXCell). Four days after injection, mice were bled to confirm depletion. hESCs were transplanted into these mice 5d after the initial depleting antibody injection. Depleting antibodies were administered weekly throughout the teratoma assay.

Pizzato H.A., Alonso-Guallart P., Woods J., Johannesson B., Connelly J.P., Fehniger T.A., Atkinson J.P., Pruett-Miller S.M., Monsma FJ J.r, & Bhattacharya D. (2023). Engineering Human Pluripotent Stem Cell Lines to Evade Xenogeneic Transplantation Barriers. bioRxiv.

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Abscopal Effect of Intratumoral CDG in Mice

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MOC1 (2 × 10⁶) or MOC2 (1 × 10⁵) cells were injected subq into the flanks of WT B6 mice. Bilateral tumors were implanted for abscopal experiments. All tumors were engrafted to at least 0.1 cm³ (6–7 mm diameter) before treatment. Mice were weighed and tumors were measured three times weekly. Intratumoral injections of R,R-CDG in HBSS (100 µL volume) were directed at the deep aspect of the tumor. Control mice received HBSS alone. PD-L1 mAb (clone 10F.9G2, rat IgG2b, BioXCell) or rat IgG isotype control antibody treatments were performed via intraperitoneal (IP) injection (200 µg/injection). In some experiments, C57BL/6J-Tmem173^gt/J STING-deficient mice (Jackson Laboratories) were used. In other experiments, cellular or cytokine depletions were performed via IP injection of CD8 (clone YTS169.4), IFNγ (clone XMG1.2) or TNFα (clone XT3.11) mAbs (200 µg/injection; BioXCell).

Moore E., Clavijo P.E., Davis R., Cash H., Van Waes C., Kim Y, & Allen C. (2016). Established T-cell inflamed tumors rejected after adaptive resistance was reversed by combination STING activation and PD-1–pathway blockade. Cancer immunology research, 4(12), 1061-1071.

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Immunological Profiling of Tumor Microenvironment

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Recombinant human interferon gamma (IFNγ; catalog no. 570206) and antibodies to human HLA-ABC (catalog no. 311404) and PD-L1 (catalog no. 329706) were obtained from Biolegend. The antibody for intracellular calreticulin was obtained from R&D Systems (catalog no. IC3898R), and the antibodies to TAP1, TAP2, LMP2, ERp57 were obtained from Abcam (catalog numbers ab83817, ab180611, ab190350, and ab13506, respectively). Pharmaceutical grade cisplatin was obtained from the veterinary pharmacy at NIH. Antibodies for in vivo mouse treatments specific for PD-L1 (clone 10F.9G2), PD-1 (clone RMP1-14), NK1.1 (clone PK136) and CD8 (clone YTS 169.4) were from BioXCell. Fluorescent-conjugated flow cytometry antibodies for mouse tumor experiments were obtained from eBioscience (CD137/41BB, catalog no. 46-1371-80) or Biolegend: CD8 (catalog no. 100712), CD45.2 (catalog no. 109806), CD80 (catalog no. 104721), CD11b (catalog no. 101211), CD11c (catalog no. 117307), CD107a (catalog no. 121619), Ly6G (catalog no. 127623), Ly6C (catalog no. 128017), and H-2Kb/H-2Db (catalog no. 114611).

Tran L., Allen C.T., Xiao R., Moore E., Davis R., Park S.J., Spielbauer K., Van Waes C, & Schmitt N.C. (2017). Cisplatin Alters Antitumor Immunity and Synergizes with PD-1/PD-L1 Inhibition in Head and Neck Squamous Cell Carcinoma. Cancer immunology research, 5(12), 1141-1151.

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