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Anti cd8α clone 2

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Anti-CD8α (clone 2.43) is a monoclonal antibody that recognizes the CD8α chain, a co-receptor expressed on the surface of cytotoxic T cells and a subset of NK cells. The antibody is commonly used in research applications for the identification and isolation of CD8+ T cells.

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Lab products found in correlation

Anti cd4 clone gk1, by BioXCell (7 mentions) Anti nk1.1 clone pk136, by BioXCell (3 mentions) Clone ltf 2, by BioXCell (2 mentions) Rat igg1, by BioXCell (1 mentions) Anti asialo gm1 antiserum, by Fujifilm (1 mentions) Rat igg2b isotype control clone ltf 2, by BioXCell (1 mentions) Anti cd4 antibodies clone gk1, by BioXCell (1 mentions) Isotype control antibody clone ltf 2, by BioXCell (1 mentions) αnk1.1 clone pk136, by BioXCell (1 mentions) Anti pd l1 clone 10f 9g2, by BioXCell (1 mentions) Anti cd4 clone gk1, by BioLegend (1 mentions) Anti pdca1 clone 927, by BioLegend (1 mentions) Clophosome a, by Formumax Scientific (1 mentions) Clone afs98, by BioXCell (1 mentions) Anti ly6g clone 1a8, by BioXCell (1 mentions) Clone trfk5, by BioXCell (1 mentions) αpd l1 clone 10f 9g2, by BioXCell (1 mentions)

11 protocols using anti cd8α clone 2

1

T Cell and NK Cell Depletion

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To deplete NK cells, CD4+ T cells and CD8+ T cells, tumour-bearing mice were intraperitoneally injected with anti-NK1.1 (clone PK136, BioXCell), anti-CD4 (clone GK1.5, BioXCell), anti-CD8-α (clone 2.43, BioXCell) or isotype control (RatIgG1,BioXcell) antibodies at an initial dose of 400 mg 1 d before treatment, followed by 200 mg every 3 d. Depletion of CD8+ T cells, CD4+ T cells and NK cells was confirmed using flow cytometry analysis of peripheral blood mononuclear cells.
Wang F., Su H., Xu D., Dai W., Zhang W., Wang Z., Anderson C.F., Zheng M., Oh R., Wan F, & Cui H. (2020). Tumour sensitization via the extended intratumoural release of a STING agonist and camptothecin from a self-assembled hydrogel. Nature biomedical engineering, 4(11), 1090-1101.
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2

Influenza Infection and Anti-CD8α Treatment

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Virus stocks of influenza A/X31(H3N2) and influenza A/PR/8 (H1N1) were produced in embryonated eggs and diluted in PBS. For virus infections, mice were anesthetized by intraperitoneal injection of a mixture of ketamine (100 mg/kg of body weight) and xylazine (5 mg/kg) before intranasal administration of either a sublethal dose at day 0 of influenza A virus/X31(H3N2) at 10 p.f.u. (LD50= 400 p.f.u.) or a lethal dose at day 35 of influenza A virus/PR/8 (H1N1) at 75 p.f.u. (LD50= 20 p.f.u.) in a volume of 50 μL. Sublethal dose was determined by lowest dose allowing mouse infection as determined by ≈10% weight loss and by viral lung titers. Lethal dose was determined as a dose of virus that kills 100% of naïve WT mice. Mice were monitored daily for clinical signs of illness, and body weights were recorded daily for 14 days. Upon reaching 75% of initial body weight, animals were humanely euthanized.
Anti-CD8α (clone 2.43, BioXcell) or control antibody (clone LTF-2, BioXcell) were injected into isoflurane anesthetized mice both intraperitoneally and intranasally every two days from day 33.
Barbet G., Nair-Gupta P., Schotsaert M., Yeung S.T., Moretti J., Seyffer F., Metreveli G., Gardner T., Choi A., Tortorella D., Tampé R., Khanna K.M., García-Sastre A, & Blander J.M. (2021). TAP dysfunction in dendritic cells enables non-canonical cross-presentation for T cell priming. Nature immunology, 22(4), 497-509.
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3

Influenza Infection and Anti-CD8α Treatment

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Virus stocks of influenza A/X31(H3N2) and influenza A/PR/8 (H1N1) were produced in embryonated eggs and diluted in PBS. For virus infections, mice were anesthetized by intraperitoneal injection of a mixture of ketamine (100 mg/kg of body weight) and xylazine (5 mg/kg) before intranasal administration of either a sublethal dose at day 0 of influenza A virus/X31(H3N2) at 10 p.f.u. (LD50= 400 p.f.u.) or a lethal dose at day 35 of influenza A virus/PR/8 (H1N1) at 75 p.f.u. (LD50= 20 p.f.u.) in a volume of 50 μL. Sublethal dose was determined by lowest dose allowing mouse infection as determined by ≈10% weight loss and by viral lung titers. Lethal dose was determined as a dose of virus that kills 100% of naïve WT mice. Mice were monitored daily for clinical signs of illness, and body weights were recorded daily for 14 days. Upon reaching 75% of initial body weight, animals were humanely euthanized.
Anti-CD8α (clone 2.43, BioXcell) or control antibody (clone LTF-2, BioXcell) were injected into isoflurane anesthetized mice both intraperitoneally and intranasally every two days from day 33.
Barbet G., Nair-Gupta P., Schotsaert M., Yeung S.T., Moretti J., Seyffer F., Metreveli G., Gardner T., Choi A., Tortorella D., Tampé R., Khanna K.M., García-Sastre A, & Blander J.M. (2021). TAP dysfunction in dendritic cells enables non-canonical cross-presentation for T cell priming. Nature immunology, 22(4), 497-509.
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4

Immune Cell Depletion Protocol

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Mice were treated weekly with 200 µg anti-CD4 antibodies (clone GK1.5, Bio X Cell, NH, USA), 200 µg anti-CD8α (clone 2.43, Bio X Cell), 10 µL anti-asialo GM1 antiserum (Wako Pure Chemical Industries, Osaka, Japan), or rat IgG2b isotype control (clone LT F-2, Bio X Cell) by intraperitoneal injection 3 days prior to irradiation. CD8+, CD4+, T cells, and NK cell depletion were confirmed by flow cytometry.
Yoon Y.N., Lee E., Kwon Y.J., Gim J.A., Kim T.J, & Kim J.S. (2022). PI3Kδ/γ inhibitor BR101801 extrinsically potentiates effector CD8+ T cell-dependent antitumor immunity and abscopal effect after local irradiation. Journal for Immunotherapy of Cancer, 10(3), e003762.
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5

Cellular Subset Depletion Protocol

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For cellular subsets depletion experiments, mice were injected intraperitoneally with 400 μg depletion antibody twice weekly beginning 1 day before therapy. The depletion antibody are as follows: anti-CD4 (clone GK1.5, BioXCell), anti-CD8α (clone 2.43, BioXCell), anti-NK1.1 (clone PK136, BioXCell) antibodies and isotype control antibody (clone LTF-2, BioXCell) 16 (link). The depletion efficacy was confirmed by flow cytometry of PBMC on day 14 (Figure S9).
Yu X., Yu J., Dai H., Deng C., Sun X., Long S., Jiang Z., Jin H., Guan Z, & Yang Z. (2022). Novel formulation of c-di-GMP with cytidinyl/cationic lipid reverses T cell exhaustion and activates stronger anti-tumor immunity. Theranostics, 12(15), 6723-6739.
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6

Depletion of Immune Cells in Murine Studies

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For CD4 and CD8 depletion, mice were injected intraperitoneally with 200 μg of anti-CD4 (clone GK1.5, BioXCell) or 200 μg of anti-CD8α (clone 2.43, BioXCell), and repeated doses were administered to achieve continuous depletion. For antibody-based treatments, 200 μg of anti-PD-L1 (clone 10 F.9G2, BioXCell) was injected intraperitoneally twice a week for a total of four doses. NK cells were depleted with repeated doses of 100 µg of αNK1.1 (clone PK136, BioXCell).
Moreo E., Jarit-Cabanillas A., Robles-Vera I., Uranga S., Guerrero C., Gómez A.B., Mata-Martínez P., Minute L., Araujo-Voces M., Felgueres M.J., Esteso G., Uranga-Murillo I., Arias M., Pardo J., Martín C., Valés-Gómez M., del Fresno C., Sancho D, & Aguiló N. (2023). Intravenous administration of BCG in mice promotes natural killer and T cell-mediated antitumor immunity in the lung. Nature Communications, 14, 6090.
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7

Selective Cell Depletion for Vaccine Evaluation

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An intraperitoneal injection with 500 µg of anti-PDCA1 (clone 927, BioLegend) or intravenous injection with 100 µg of anti-CD4 (clone GK1.5, BioLegend) was performed to deplete pDCs or CD4+ cells 1 day before immunization with the vaccine. To deplete CD8+ cells, an intraperitoneal injection with 100 µg of anti-CD8α (clone 2.43, Bio X cell, Lebanon, NH) was performed at 15, 17, 19, and 21 days after the inoculation of tumor. To deplete macrophages, 200 µL of clophosome-A (FormuMax Scientific, Inc., Sunnyvale, CA) was intravenously administered on days 1 and 6 before immunization with the vaccine.
Nakagawa T., Tanino T., Onishi M., Tofukuji S., Kanazawa T., Ishioka Y., Itoh T., Kugimiya A., Katayama K., Yamamoto T., Nagira M, & Ishii K.J. (2021). S-540956, a CpG Oligonucleotide Annealed to a Complementary Strand With an Amphiphilic Chain Unit, Acts as a Potent Cancer Vaccine Adjuvant by Targeting Draining Lymph Nodes. Frontiers in Immunology, 12, 803090.
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8

Cellular Depletion Strategies for Immunotherapy

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Cellular subsets were depleted by administering 400 μg of depleting antibody i.p. twice weekly beginning one day prior to therapy as indicated: CD8 T-cells with anti-CD8α (clone 2.43, BioXCell), CD4 T-cells with anti-CD4 (clone GK1.5, BioXCell), NK cells with anti-NK1.1 (clone PK136, BioXCell), neutrophils with anti-Ly-6G (clone 1A8, BioXCell), with the exception of CSF1R and IL-5, which were depleted using 300 μg (clone AFS98, BioXCell) every other day and 1 mg (clone TRFK5, BioXCell) weekly respectively.9 (link) Cellular depletions of CD8 T cells, CD4 T cells, neutrophils, and NK cells were confirmed by flow cytometry of PBMC (Supplementary Fig. 7).
Moynihan K.D., Opel C.F., Szeto G.L., Tzeng A., Zhu E.F., Engreitz J.M., Williams R.T., Rakhra K., Zhang M.H., Rothschilds A.M., Kumari S., Kelly R.L., Kwan B.H., Abraham W., Hu K., Mehta N.K., Kauke M.J., Suh H., Cochran J.R., Lauffenburger D.A., Wittrup K.D, & Irvine D.J. (2016). Eradication of large established tumors in mice by combination immunotherapy that engages innate and adaptive immune responses. Nature medicine, 22(12), 1402-1410.
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9

Depletion of T Cell Subsets

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Cellular subsets were depleted by administering 300 µg of depleting antibody intraperitoneally twice weekly starting one day prior to immunotherapy: CD8+ T-cells with anti-CD8α, clone 2.43 (BioXCell), and CD4+ T-cells with anti-CD4, clone GK1.5 (BioXCell). Cellular depletion of CD8+ and CD4+ T-cells were confirmed by flow cytometry of PBMC blood levels (Figure S3). Samples were measured by FACSCantoII. FlowJo software was used for analysis.
Caisova V., Li L., Gupta G., Jochmanova I., Jha A., Uher O., Huynh T.T., Miettinen M., Pang Y., Abunimer L., Niu G., Chen X., Ghayee H.K., Taïeb D., Zhuang Z., Zenka J, & Pacak K. (2019). The Significant Reduction or Complete Eradication of Subcutaneous and Metastatic Lesions in a Pheochromocytoma Mouse Model after Immunotherapy Using Mannan-BAM, TLR Ligands, and Anti-CD40. Cancers, 11(5), 654.
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10

In vivo CD4/CD8 Depletion and PD-L1 Blockade

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For CD4 and CD8 depletion, mice were injected intraperitoneally with 300 µg of anti-CD4 (clone GK1.5, BioXCell) or 300 µg of anti-CD8α (clone 2.43, BioXCell) 3 days before tumor challenge or the day before intravesical treatment. Repeated doses were administered to achieve continuous depletion if needed. For antibody-based treatment, 200 µg of αPD-L1 (clone 10F.9G2, BioXCell) was injected intraperitoneally two times a week.
Moreo E., Uranga S., Picó A., Gómez A.B., Nardelli-Haefliger D., del Fresno C., Murillo I., Puentes E., Rodríguez E., Vales-Gómez M., Pardo J., Sancho D., Martín C, & Aguilo N. (2022). Novel intravesical bacterial immunotherapy induces rejection of BCG-unresponsive established bladder tumors. Journal for Immunotherapy of Cancer, 10(7), e004325.
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