Be0004 1

Manufactured by BioXCell

The BE0004-1 is a laboratory centrifuge designed for general-purpose applications. It features a compact and durable construction, accommodating up to 4 sample tubes or 2 microplates. The centrifuge operates at adjustable speeds up to 6,000 rpm, providing reliable separation of samples in a range of scientific workflows.

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12 protocols using be0004 1

In Vivo Tumor Modeling in Mice

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All animal studies were reviewed and approved by the Institutional Animal Care and Use Committee of the Suzhou Institute of Systems Medicine. Female C57BL/6 and athymic nude BALB/c mice (nu/nu) (6–8 weeks) were purchased from Beijing Vital River Company. Mice were randomly divided into the indicated groups (5 mice/group) before inoculation. Cancer cells (2 × 10⁶ cells in 100 μl PBS per mouse) were subcutaneously implanted. Tumor size was monitored using calipers 2–3 times per week and calculated by multiplying the length by the width. In some experiments, CD8⁺ T cells were depleted by injecting 200 µg/mouse anti-CD8 antibodies (BE0004-1, BioXCell) intravenously at the indicated time points. Tumors were harvested on day 7–12 postimplantation for RNA sequencing, flow cytometric analysis, immunofluorescence staining, and ELISpot analysis. Tumor growth curves are depicted with the error bars indicating the mean ± SD at each time point. Cancer cells were injected via the tail vein, and mortality was recorded every 24 h. Kaplan–Meier survival curves were also plotted. Animals were euthanized with CO₂ when the tumor volume reached 300 mm².

Liu W., Gao M., Li L., Chen Y., Fan H., Cai Q., Shi Y., Pan C., Liu J., Cheng L.S., Yang H, & Cheng G. (2021). Homeoprotein SIX1 compromises antitumor immunity through TGF-β-mediated regulation of collagens. Cellular and Molecular Immunology, 18(12), 2660-2672.

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Antibody Depletion of T-cells for ZIKV

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T-cell depletion was achieved by two consecutive injections of 500 µg of anti-CD4 (Bio X Cell; BE0003-1) and anti-CD8 (Bio X Cell; BE0004-1) monoclonal antibodies i.p. at 2 days and 24 h before ZIKV challenge.^{41 (link)} Antibody depletions were more than 95% efficient as determined by flow cytometry.

Li G., Adam A., Luo H., Shan C., Cao Z., Fontes-Garfias C.R., Sarathy V.V., Teleki C., Winkelmann E.R., Liang Y., Sun J., Bourne N., Barrett A.D., Shi P.Y, & Wang T. (2019). An attenuated Zika virus NS4B protein mutant is a potent inducer of antiviral immune responses. NPJ Vaccines, 4, 48.

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Oncolytic virus therapy with immune checkpoint blockade

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When inoculated tumor volumes reached ~50 mm³, tumor-bearing mice were randomized into control or treatment groups, and unilateral intratumorally injected with viral suspension in phosphate buffered saline (PBS) (5×10⁶ pfu/mouse) or PBS along every third day. For combined therapy, CD40 antibody (100 µg/dose, BE0016-2, BioXCell), PD-1 antibody (100 µg/dose, BE0146, BioXCell) or isotype antibody (BE0089, BioXCell) was injected intraperitoneally on the day of viruses or PBS injection. For CD8⁺ T cells depletion and cytokines neutralization experiments, CD8α antibody (250 µg/dose, BE0004-1, BioXCell), interleukin (IL)-12 antibody (500 µg/dose, BE0233, BioXCell), IFN-γ antibody (250 µg/dose, BE0055, BioXCell), or indicated isotype antibodies (BE0088, BE0089, or BE0090, BioXCell) was injected intraperitoneally 24 hours prior to each viral treatment and administrated in 7-day intervals after the last oHSVs treatment, respectively.

Wang R., Chen J., Wang W., Zhao Z., Wang H., Liu S., Li F., Wan Y., Yin J., Wang R., Li Y., Zhang C., Zhang H, & Cao Y. (2022). CD40L-armed oncolytic herpes simplex virus suppresses pancreatic ductal adenocarcinoma by facilitating the tumor microenvironment favorable to cytotoxic T cell response in the syngeneic mouse model. Journal for Immunotherapy of Cancer, 10(1), e003809.

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Modulating CEBPA for Tumor Immunotherapy

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MTL-CEBPA, a liposomal nanoparticle encapsulating a small activating RNA (saRNA) for C/EBPa (CEBPA-51), and its control liposomal nanoparticle with a non-specific oligonucleotide (NOV-FLUC) encapsulating siFLUC were supplied by MiNA Therapeutics Ltd, London, United Kingdom. MTL-CEBPA or NOV-FLUC was intravenously injected to the tumor-bearing mice twice per week at 3 mg/kg. For the T cell depletion study, intraperitoneal injection with 100 μg of anti-mouse CD8a antibody (BioXcell, BE0004-1) or rat IgG2a isotype control antibody (BioXcell) was started 2 days before tumor injection and repeated twice a week for 2 weeks. Anti-mouse CTLA-4 antibody (BioXcell, BE0164) or mouse IgG2b isotype control antibody was intraperitoneally injected to the tumor-bearing mice 100 μg per mouse on Days 10, 17 and 24. Celecoxib, selective cox2 inhibitor (Selleck Chemicals) was suspended in 0.5% methylcellulose and orally treated at 50 mg/kg to the tumor-bearing mice every day. Lipofermata was dissolved in DMSO and diluted in 30% (v/v) Kolliphor, and then subcutaneously injected to the tumor-bearing mice at 2 mg/kg twice a day.

Hashimoto A., Sarker D., Reebye V., Jarvis S., Sodergren M.H., Kossenkov A., Sanseviero E., Raulf N., Vasara J., Andrikakou P., Meyer T., Huang K.W., Plummer R., Chee C.E., Spalding D., Pai M., Khan S., Pinato D.J., Sharma R., Basu B., Palmer D., Ma Y.T., Evans J., Habib R., Martirosyan A., Elasri N., Reynaud A., Rossi J.J., Cobbold M., Habib N.A, & Gabrilovich D.I. (2021). Upregulation of C/EBPα Inhibits Suppressive Activity of Myeloid Cells and Potentiates Antitumor Response in Mice and Patients with Cancer. Clinical Cancer Research, 27(21), 5961-5978.

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CD8+ T-cell Depletion in Glioma Model

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Animals were treated with either anti-mouse CD8α (BE0004-1, BioXCell) or anti-rat IgG2a isotype control, anti-trinitrophenol (BE0089, BioXCell). The antibody concentration given to each mouse was 2 mg/mL diluted in PBS at a final volume of 100 μL administered via intraperitoneal injection one week prior to glioma induction and continued twice per week in order to achieve consistent CD8⁺ T-cell depletion. Depletion was monitored at three time points starting from intracranial virus injection, 3 weeks apart.

Kane J.R., Zhao J., Tsujiuchi T., Laffleur B., Arrieta V.A., Mahajan A., Rao G., Mela A., Dmello C., Chen L., Zhang D.Y., González-Buendia E., Lee-Chang C., Xiao T., Rothschild G., Basu U., Horbinski C., Lesniak M.S., Heimberger A.B., Rabadan R., Canoll P.D, & Sonabend A.M. (2020). CD8+ T-cell-mediated immunoediting influences genomic evolution and immune evasion in murine gliomas. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(16), 4390-4401.

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Immunomodulation Strategies for Cancer

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Anti‐mouse PD‐1 antibody (BE0273; Bioxcell) was administered via intraperitoneal injection at a dose of 200 µg per mouse. Rat immunoglobulin G 2a (BE0089, Bioxcell) was used as a control twice per week for four weeks (Figure 3A). To deplete CD8 T and CD4 T cells, 200 µg of anti‐mouse CD8α (BE0004‐1; Bioxcell), 200 µg of anti‐mouse CD4 (BE0003‐3; Bioxcell), or 200 µg of rat IgG2a (BE0089; Bioxcell) as an isotype control were administered via intraperitoneal injection shown in Figure 6E. Depletion efficiency was determined by IHC using anti‐CD8 antibody (14.0808.82; eBioscience) and anti‐CD4 antibody (D7D2Z; Cell Signaling Technology).
To neutralize IFN‐γ, mice were intraperitoneally injected with antibodies against IFN‐γ (1.25 mg kg⁻¹, BE0312; BioXcell). Polyclonal Armenian hamster IgG (BE0091; BioXcell) was used as a control (Figure 6E).

Vaziri F., Setayesh T., Hu Y., Ravindran R., Wei D, & Wan Y.Y. (2024). BCG as an Innovative Option for HCC Treatment: Repurposing and Mechanistic Insights. Advanced Science, 11(14), 2308242.

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S. aureus Bioparticle Therapy in LLC Tumors

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A total of 1×10⁵ LLC tumor cells were inoculated into ear pinnae of C57BL/6 mice. Once tumors were visible, CD8 T cells were depleted by intraperitoneal injection of 250 μg anti-mouse CD8 clone 53–6.7 (Bio X Cell, catalog no. BE0004–1 RRID:AB_1107671) or isotype clone IgG2a (Bio X Cell, catalog no. BE0089 RRID:AB_1107769). Twenty-four hours later, 20 × 10⁶S. aureus bioparticles or vehicle control, were injected into tumors. Subsequent anti-CD8 antibodies were administered every 3 days and S. aureus bioparticles (or vehicle control) every 2 days. A total of five anti-CD8 or isotype injections were administered and S. aureus bioparticles (or vehicle) were given until mice reached ethical endpoints.

Yam A.O., Bailey J., Lin F., Jakovija A., Youlten S.E., Counoupas C., Gunzer M., Bald T., Woodruff T.M., Triccas J.A., Goldstein L.D., Gallego-Ortega D., Grey S.T, & Chtanova T. (2023). Neutrophil Conversion to a Tumor-Killing Phenotype Underpins Effective Microbial Therapy. Cancer Research, 83(8), 1315-1328.

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Depleting CD4+ and CD8+ T cells

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For depletion of CD4⁺ and CD8⁺ T cells, mice were injected intraperitoneally weekly with rat InVivoMAb anti-mouse CD4 (GK1.5, BE0003-1, BioXcell) and rat InVivoMAb anti-mouse CD8α (53–6.7, BE0004-1, BioXcell), or isotype control IgG2b (LTF-2, BE0090, BioXcell) and IgG2a (2A3, BE0089, BioXcell) at a dose of 250 μg per mouse. These regimens resulted in > 99% depletion of CD4⁺ and CD8⁺ T cells from the peripheral blood, as evaluated by flow cytometry (supplementary figure S1a).

Li Q., Li Y., Wang Y., Xu L., Guo Y., Wang Y., Wang L, & Guo C. (2021). Oral administration of Bifidobacterium breve promotes antitumor efficacy via dendritic cells-derived interleukin 12. Oncoimmunology, 10(1), 1868122.

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Antigen-specific Immunotherapy against Melanoma

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Female C57BL/6 mice were subcutaneously
inoculated with B16F10 cells (2 × 10⁵/mouse). After
the tumor size reached about 100 mm³, the mice were divided
into 2 treatment groups, including αCD8 + MSA-2-Pt
and IgG + MSA-2-Pt. The αCD8 + MSA-2-Pt
group was treated with antimouse CD8α (α-CD8, Bioxcell,
BE0004-1, 200 μg per mouse, i.p.) + MSA-2-Pt (i.t. 150 μg/mouse, n = 10). IgG + MSA-2-Pt group were i.t. injected
with isotype control antibody (Bioxcell, Catalog # BP0090, 200 μg
per mouse,i.p.) + MSA-2-Pt (i.t. 150 μg/mouse, n = 10) for three doses.

Wang M., Cai Y., He T., Zhang Y., Yi L., Li W, & Zhou P. (2024). Antitumor Effect of Platinum-Modified STING Agonist MSA-2. ACS Omega, 9(2), 2650-2656.

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Multimodal Anticancer Therapy in Mice

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Adapalene (MedChemExpress, HY-B0091) and Bexarotene (MedChemExpress, HY-14171) were administered by oral gavage three days/week at the dose of 20 mg/kg and daily at the dose of 20 mg/kg, respectively. Adapalene was resuspended in DMSO 10%, PEG-400 40%, Tween-80 10%. Docetaxel (TEVA, RDCXVR100800) was administered weekly at the dose of 10 mg/kg by intraperitoneal injection.¹³ (link)^,³⁹ (link)^,⁴² (link) For the in vivo depletion of NK cells, mice were treated with either 50 μg of anti-NK1.1 (clone BE0036, BioXCell), 100 μg Asialo GM1 Polyclonal Antibody (Invitrogen # 16-6507-39), or the isotype control (C1.18.4, BioXCell) on Mondays, Wednesdays, and Fridays. For the in vivo depletion of CD8 T cells, mice were treated with 200 ug of anti-CD8a (clone 53-6.7, Bio X Cell BE0004-1, RRID AB_1125541) or the isotype control (LTF-2, Bio X Cell BE0090, RRID AB_1107780) on Mondays and Thursdays. The respective RRID of each compound is listed in the key resources table. Mice were monitored for suffering, distress, or weight loss by measuring the total body weight weekly and assessing the behavioral changes every day for the entire duration of the experiment.

Colucci M., Zumerle S., Bressan S., Gianfanti F., Troiani M., Valdata A., D’Ambrosio M., Pasquini E., Varesi A., Cogo F., Mosole S., Dongilli C., Desbats M.A., Contu L., Revankdar A., Chen J., Kalathur M., Perciato M.L., Basilotta R., Endre L., Schauer S., Othman A., Guccini I., Saponaro M., Maraccani L., Bancaro N., Lai P., Liu L., Pernigoni N., Mele F., Merler S., Trotman L.C., Guarda G., Calì B., Montopoli M, & Alimonti A. (2024). Retinoic acid receptor activation reprograms senescence response and enhances anti-tumor activity of natural killer cells. Cancer Cell, 42(4), 646-661.e9.

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