Anti cd8 clone yts169.4

Manufactured by BioXCell

Sourced in United States

Anti-CD8 (clone YTS169.4) is a monoclonal antibody that recognizes the CD8 surface antigen. CD8 is a co-receptor found on cytotoxic T cells and a subset of natural killer cells. The Anti-CD8 (clone YTS169.4) antibody can be used in various applications to identify and study CD8-positive cells.

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Anti-CD8 (28 citations) Anti-CD8 antibody (clone YTS169.4; (3 citations) Anti-CD8 (YTS169.4) (2 citations)

18 protocols using anti cd8 clone yts169.4

Severe T Cell Lymphopenia Induction

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To induce severe T cell lymphopenia 100 μg anti CD4 (clone YTS 191) and 50 μg of anti CD8 (clone YTS 169.4), both BioXcell were given intraperitoneally. Where stated, groups of mice were given 300 μg mAb to IL-7R (clone A7R34; a hybridoma prepared in-house) intraperitoneally every second day over the 14 days experimental time-course. The anti CD25 antibody (clone PC61) was purchased from BioXcell and a single dose of 500 μg per mouse was injected intraperitoneally.

Knipper J.A., Wright D., Cope A.P., Malissen B, & Zamoyska R. (2020). PTPN22 Acts in a Cell Intrinsic Manner to Restrict the Proliferation and Differentiation of T Cells Following Antibody Lymphodepletion. Frontiers in Immunology, 11, 52.

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Immune Cell Depletion in Tumor Models

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For macrophage-depletion experiments, mice bearing NB9464 tumors were treated with 50 mg/kg anti-CSF-1R antibody (clone AFS 98, BioXcell), administered intraperitoneally (ip) every alternate day until tumors were harvested. For B-cell depletion experiments, mice received tail vein injection of 250 µg of ULTRA-LEAF purified anti-mouse CD-20 (clone SA271G2, Bio Legend). For CD8-depletion experiments, mice bearing NB9464 tumors were treated with 200µg of anti-CD8 (clone YTS169.4) from Bio-X-Cell administered ip on day 7, 10 and 13 of tumor inoculation as described before (41 (link)).

Rohila D., Park I.H., Pham T.V., Jones R., Tapia E., Liu K.X., Tamayo P., Yu A., Sharabi A.B, & Joshi S. (2023). Targeting macrophage Syk enhances responses to immune checkpoint blockade and radiotherapy in high-risk neuroblastoma. Frontiers in Immunology, 14, 1148317.

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Induction of CD8+ Tissue-Resident Memory T Cells

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To induce CD8⁺ T_RM cells, 5 × 10⁴ WT- or Itga1^−/−⁻ CD8⁺ OT-I T cells in 200 μL PBS were adoptively transferred i.v. into recipient mice. Recipient mice were then injected intradermally twice (separated by three days) with 3 μg of the plasmid, pODCAGGS (Mani et al., 2019 (link)) which encodes full length chicken ovalbumin, kindly provided by T. Mempel. At least 28 d after DNA vaccination, mice were injected i.p. with 500 μg anti-CD4 (GK1.5; Biolegend) and 300 μg anti-CD8 (clone YTS169.4; BioXCell) antibodies to deplete circulating T cells. After 4 d, mice were bled and stained for CD4 and CD8 to confirm T cell depletion. Then, mice were infected with 10⁶ pfu HSV-OVA at the site of DNA vaccination. After 4 d, a 1 cm² piece of skin surrounding the infection site was isolated, minced and placed in 1 mL of DMEM (Life Technologies) for determination of viral titers by plaque assay.

Bromley S.K., Akbaba H., Mani V., Mora-Buch R., Chasse A.Y., Sama A, & Luster A.D. (2020). CD49a Regulates Cutaneous Resident Memory CD8+ T Cell Persistence and Response. Cell reports, 32(9), 108085.

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Macrophage and CD8+ T Cell Depletion in Murine Tumor Models

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For macrophage depletion experiments, mice bearing KPC1245 tumors were treated with 50 mg/kg of anti-CSF1R antibody (clone AFS 98, BioXcell), administered intraperitoneally, every alternate day, until tumors were harvested on day 15. For CD8 depletion experiments, mice with KPC1245 tumors were treated with 200 μg of anti-CD8 (clone YTS169.4) or an isotype rat IgG2b control (LTF-2) from Bio-X-Cell administered intraperitoneally on day −3, 0, 3, 6, and 10 of tumor inoculation as described previously (20 (link)).

Rohila D., Park I.H., Pham T.V., Weitz J., Hurtado de Mendoza T., Madheswaran S., Ishfaq M., Beaman C., Tapia E., Sun S., Patel J., Tamayo P., Lowy A.M, & Joshi S. (2023). Syk Inhibition Reprograms Tumor-Associated Macrophages and Overcomes Gemcitabine-Induced Immunosuppression in Pancreatic Ductal Adenocarcinoma. Cancer Research, 83(16), 2675-2689.

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Investigating Immune Checkpoint Inhibitor Therapy in Liver Inflammation

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Anti-PD-1 (clone RMP1-14), anti-CTLA-4 (clone 9H10), anti-CD4 (clone GK1.5) and anti-CD8 (clone YTS169.4) monoclonal antibodies (mAbs) were purchased from Bio X Cell, diluted in sterile phosphate-buffered saline (PBS) and injected intraperitoneally (i.p.) at a concentration of 200 µg/mouse.
Single anti-PD-1 and anti-CTLA-4 or combination anti-PD-1/CTLA-4 (“CPI”) or sterile PBS as control (“PBS”) were given every 2 days starting on day 0 (D0). On D1, 20 µg/mouse of TLR9-L (CpG oligodeoxynuleotide 1668: 5-S-TCCATGACGTTC CTGATGCT-3) (TIB Molbiol, Germany) was administered i.p. to prime hepatic inflammation. Mice were sacrificed, and blood and liver tissue were collected on D1, D4, D7, D10 and D14 as described in online supplemental methods.
Anti-CD4 or anti-CD8 mAbs were given on D-2 and D-1 before and on D3 after the first CPI injection. 100 mg/kg/day of CVC mesylate (MedchemExpress, USA) or vehicle control was administered in drinking water1 (link) 4 days before the first CPI injection (“prophylactic”) or2 (link) 4 days following the first CPI injection (“therapeutic”) and for the whole duration of the time course.25 (link) Mice were sacrificed on D7 or D10 and blood and liver tissue were harvested.

Gudd C.L., Mitchell E., Atkinson S.R., Mawhin M.A., Turajlic S., Larkin J., Thursz M.R., Goldin R.D., Powell N., Antoniades C.G., Woollard K.J., Possamai L.A, & Triantafyllou E. (2024). Therapeutic inhibition of monocyte recruitment prevents checkpoint inhibitor-induced hepatitis. Journal for Immunotherapy of Cancer, 12(4), e008078.

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Selective Depletion of Immune Cells

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CD4⁺ T cells, CD8⁺ T cells, and NK cells were depleted by intraperitoneal (i.p.) injection of 200 μg of anti-CD4 (clone GK1.5), anti-CD8 (clone YTS 169.4), and anti-NK1.1 (clone PK136) antibodies respectively in antibody dilution buffer pH 7.0 (Bio X Cell). Cellular depletion was confirmed by flow cytometry analysis of CD4⁺ T cells, CD8⁺ T cells and NK cells in the blood samples of mice within 24 hours after the first depletion antibody injection. The next day, mice were challenged with 5 × 10⁵ LL/2-tdTomato/Luc cells and another four doses of cellular depletion antibodies were given every four to five days post tumor challenge to deplete newly formed lymphocytes.

Huang L., Bommireddy R., Munoz L.E., Guin R.N., Wei C., Ruggieri A., Menon A.P., Li X., Shanmugam M., Owonikoko T.K., Ramalingam S.S, & Selvaraj P. (2021). Expression of tdTomato and luciferase in a murine lung cancer alters the growth and immune microenvironment of the tumor. PLoS ONE, 16(8), e0254125.

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SARS-CoV-2 Spike Protein Vaccine Evaluation

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hyACE2 knock-in mice were vaccinated via intramuscular injection into the gastrocnemius muscle with 10 μg and 5 μg (28 days later) of SARS-CoV-2 full-length spike (S-2P) mRNA-LNP or luciferase mRNA-LNP^{54 (link)}. After 28 days, mice received an immunization boost with 5 μg SARS-CoV-2 S-2P mRNA-LNP or luciferase mRNA-LNP. SARS-Cov-2 S-2P mRNA vaccines were designed based on the SARS-CoV-2 full-length spike (S) protein sequence with K986P and V987P amino acids substitutions (Wuhan-Hu-1, GenBank: MN908947.3)^{38 (link)}.
For the experiments described in Fig. 6, mice were injected intravenously with 200 μg per mouse of anti-CD4 (clone GK1.5, BioXcell), anti-CD8 (clone YTS169.4, BioXcell) or both, three times 2 days apart. In addition, a group of mice was injected intravenously with 250 μg per mouse of anti-mouse IFN-γ (clone XMG1.2, BioXcell) two times, 4 h before and 3 days after the viral challenge.
In the experiments described in Fig. 4, mice were treated with 50 μg per mouse of Treg-Protector (anti-ARTC2 nanobody; clone: S + 16a; BioLegend, 149802) by intravenous injection 30 min before sacrificing them.

Fumagalli V., Ravà M., Marotta D., Di Lucia P., Bono E.B., Giustini L., De Leo F., Casalgrandi M., Monteleone E., Mouro V., Malpighi C., Perucchini C., Grillo M., De Palma S., Donnici L., Marchese S., Conti M., Muramatsu H., Perlman S., Pardi N., Kuka M., De Francesco R., Bianchi M.E., Guidotti L.G, & Iannacone M. (2024). Antibody-independent protection against heterologous SARS-CoV-2 challenge conferred by prior infection or vaccination. Nature Immunology, 25(4), 633-643.

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Blockade of INFAR1 and T cell depletion

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For INFAR1 blockade, 1 mg of anti-IFNAR-1 antibody (clone MAR1–5A3, Bio X Cell, Lebanon, NH, USA) or isotype control was administered to mice intravenously via retroorbital injection the day prior to dying cell immunization. For T cell depletion, 350 μg of anti-CD8 (clone YTS 169.4, Bio X Cell), anti-CD4 (clone GK1.5, Bio X Cell), or Isotype control was administered to mice intravenously prior to dying cell immunization. Where indicated, an additional 150 μg of anti-CD8, anti-CD4, or Isotype control was administered on the day prior to dying cell immunization.

Chan F., Rucker A.J., Park C., Li Q.J, & Moseman E.A. (2023). Necroptosis Stimulates Interferon-Mediated Protective Anti-Tumor Immunity. Research Square.

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CD8+ T Cell Depletion Protocol

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Mice were depleted of CD8⁺ T cells by i.v. injection 200 μg anti-CD8 (clone YTS 169.4; BioXcell) on day 0 and 4 post-infection.

Rua R., Lee J.Y., Silva A.B., Swafford I.S., Maric D., Johnson K.R, & McGavern D.B. (2019). Infection drives meningeal engraftment by inflammatory monocytes that impairs CNS immunity. Nature immunology, 20(4), 407-419.

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Macrophage and CD8 Depletion in KPC1245 Tumors

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For macrophage depletion experiments, mice bearing KPC1245 tumors were treated with 50 mg/kg of anti-CSF1R antibody (clone AFS 98, BioXcell, administered intraperitoneally, every alternate day, until tumors were harvested on day 15. For CD8 depletion experiments, mice with KPC1245 tumors were treated with 200μg of anti-CD8 (clone YTS169.4) or an isotype rat IgG2b control (LTF-2) from Bio-X-Cell administered ip on day −3, 0, 3, 6 and 10 of tumor inoculation as described before (20 (link)).

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