Isqavhaahaeineagr

Manufactured by Merck Group

Sourced in Israel

Description not available

Automatically generated - may contain errors

Lab products found in correlation

Cd45.1 congenic mice, by Jackson ImmunoResearch (1 mentions) Celltrace violet, by Thermo Fisher Scientific (1 mentions) Cd4 t cell isolation kit, by STEMCELL (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions) Roswell park memorial institute (rpmi), by Corning (1 mentions) Cd4 t cell isolation kit, by Miltenyi Biotec (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) Cay10404, by Cayman Chemical (1 mentions) Facscanto, by BD (1 mentions) Ova257 264, by Merck Group (1 mentions) Siinfekl, by Merck Group (1 mentions) Ova323 339, by Merck Group (1 mentions)

5 protocols using isqavhaahaeineagr

Genetic Manipulation of Murine Dendritic Cells

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C57BL/6 mice were from Joint Ventures Sipper BK Experimental Animal Company (Shanghai, China). OT-II mice (which have transgenic expression of a T-cell antigen receptor specific for chicken ovalbumin amino acids 323–339 (OVA_(323–339)) (amino acids 323–339) (ISQAVHAAHAEINEAGR) (Sigma-Aldrich) in the context of the MHC class II molecule I-Ab), CD45.1⁺ congenic mice, and transgenic CD11c-cre mice were from The Jackson Laboratory. Mice bearing a Mettl3^fl allele (Mettl3^fl mice) were from Chinese Academy of Sciences^{53 (link)}. Mice lacking Mettl3 exon2, exon3, and exon4 specifically in DC, were generated by breeding of mettl3^fl mice with CD11c-Cre mice. All mice were maintained under pathogen-free conditions and were used at 6–8 weeks of age unless indicated otherwise. All animal experiments were carried out according to the National Institute of Health Guide for the Care and Use of Laboratory Animals, with the approval of the Scientific Investigation Board of Second Military Medical University (Shanghai, China).

Wang H., Hu X., Huang M., Liu J., Gu Y., Ma L., Zhou Q, & Cao X. (2019). Mettl3-mediated mRNA m6A methylation promotes dendritic cell activation. Nature Communications, 10, 1898.

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Dendritic Cell-Mediated T Cell Activation

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Bone marrow was obtained from the long bones of FcRγ^−/− B6 mice and plated at a density of 10⁶ cells per 6-well dishes in RPMI (Corning), 10 % FCS, 1 %l-Glutamine, streptomycin (100 mg/ml), and penicillin (100 U/ml). For the maturation of DCs, 40 ng/ml of both GM-CSF and IL-4 (PeproTech, Rocky Hill, NJ) was added to the media. Media were replaced after 3 days, and on the 6th day mature DCs were replated at a density of 5 × 10⁴ per well in U-bottom 96 wells. Two μM of DTA-1 was administered to DCs for all in vitro experiments. For DC-pulsed T cell proliferation assays, BMDC was pulsed with OVA peptides for MHC-class I (SIINFEKL—1 μg/ml) or MHC-class II (ISQAVHAAHAEINEAGR—10 μg/ml) (Sigma) and 2 μM DTA-1. After 24 h, CD8 T cells from OTI mice (CD4⁺ from OTII mice) were obtained from splenocytes using a CD8 or CD4 T cell isolation kit, respectively (StemCell Technologies, Vancouver, CA), and stained with Cell Trace Violet (Life Technologies, Carlsbad, CA) as previously described [41 ]. After 72 h, cells were transferred to V-bottom 96 wells and stained for via flow cytometry.

Miska J., Rashidi A., Chang A.L., Muroski M.E., Han Y., Zhang L, & Lesniak M.S. (2016). Anti-GITR therapy promotes immunity against malignant glioma in a murine model. Cancer immunology, immunotherapy : CII, 65(12), 1555-1567.

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Kupffer Cell-Mediated CD4+ T Cell Modulation

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FACS-sorted Kupffer cells from control or HCC-bearing mice were cultured in 96-well plates overnight. Splenocytes and lymph node cells from OT-II mice were labeled with CFSE (0.5 µM). CD4⁺ T cells were purified using a CD4⁺ T Cell Isolation Kit (Miltenyi Biotec). After 3-h-incubation with ovalbumin^323-339 peptide (ISQAVHAAHAEINEAGR, 40 ng/mL, Sigma-Aldrich) with or without LPS (400 ng/mL, Sigma-Aldrich), CFSE-labeled CD4⁺ cells were added to each well at different Kupffer cells/CD4⁺ cells ratio. For selected experiments anti-PD-L1 (10F.9G2), anti-IL-10 (JES5-16E3) (Biolegend), and CAY10404 (Cayman Chemical) were added.

Lacotte S., Slits F., Orci L.A., Meyer J., Oldani G., Delaune V., Gonelle-Gispert C., Morel P, & Toso C. (2016). Impact of myeloid-derived suppressor cell on Kupffer cells from mouse livers with hepatocellular carcinoma. Oncoimmunology, 5(11), e1234565.

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Detailed Reagents for Murine Immune Profiling

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Pertussis toxin (PT) (cat. no. P7208), peptides corresponding to residues 35–55 of rat MOG (MEVGWYRSPFSRVVHLYRNGK) and residues 323–339 (ISQAVHAAHAEINEAGR) of OVA were purchased from Sigma (Rehovot, Israel). Anti‐mouse CD4‐fluorescein isothiocyanate (FITC) (cat. no. 11‐0042‐85; clone RM4‐5), CD4‐phycoerythrin (PE) (cat. no. 12‐0042‐82; clone RM4‐5), CD8a‐PE‐cyanine 7 (Cy7) (cat. no. 25‐0081‐81; clone 53‐6.7), interferon (IFN)‐γ‐PE, interleukin (IL)‐17‐Alexa‐Fluor 647, CD25‐PE (cat. no. 12‐0251‐82; clone PC61.5), CD16/32 (cat. no. 14‐0161‐85; clone 93), forkhead box protein 3 (FoxP3)‐peridinin chlorophyll (PerCP)‐Cy5.5 (cat. no. 45‐5773‐80; clone FJK‐16s), CD3 (cat. no. 16‐0031‐85; clone 145‐2C11), CD28 (cat. no. 16‐0281‐85; clone 37.51), anti‐Ki‐67‐PE (cat. no. 12‐5698‐80; clone SoIA15), anti‐lymphocyte antigen 6C (Ly6C) Alexa‐Fluor 488 (cat. no. 53‐5932‐82; clone HK1.4), anti‐Ly6G‐allophycocyanin (APC) (cat no. 17‐9668‐82; clone 1A8‐Ly6g), anti‐CD11b‐PE (cat. no. 12‐0112‐82; clone M1/70), anti‐CD115‐APC (cat. no. 17‐1152‐82; clone AFS98) and anti‐B220‐APC (cat. no. 17‐0452‐82; clone RA3‐6B2) were all purchased from Thermo Fisher Scientific (Fremont, CA, USA.

Yacov N., Kafri P., Salem Y., Propheta‐Meiran O., Feldman B., Breitbart E, & Mendel I. (2020). MOSPD2 is a therapeutic target for the treatment of CNS inflammation. Clinical and Experimental Immunology, 201(2), 105-120.

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GABA Modulates T Cell Proliferation

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Lymphocytes were isolated from lymph nodes of OTI or OTII mice and stimulated with SIINFEKL (OVA_257–264; Sigma-Aldrich) or ISQAVHAAHAEINEAGR (OVA_323–339; Sigma-Aldrich) peptide at 5 μM for 24 hours. Cells were then labeled with 5 μM CSFE (Thermo Fisher Scientific) at 37 °C for 10 minutes. After two washes, cells were seeded in 48-well plates (1×10⁶ cells per well) and then treated with the indicated concentrations of GABA. Three days after incubation, stained cells were counted to quantify the proliferation of OTI CD8⁺ T and OTII CD4⁺ T cells using a BD FACS Canto flow-cytometry system.

Huang D., Wang Y., Thompson J.W., Yin T., Alexander P.B., Qin D., Mudgal P., Wu H., Liang Y., Tan L., Pan C., Yuan L., Wan Y., Li Q.J, & Wang X.F. (2022). Cancer Cell-Derived GABA Promotes β-Catenin-Mediated Tumor Growth and Immunosuppression. Nature cell biology, 24(2), 230-241.

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