type b cpg odn 2006  (InvivoGen)

Journal: Arthritis research & therapy

Article Title: Differential expression pattern of Bcl-2 family members in B and T cells in systemic lupus erythematosus and rheumatoid arthritis.

doi: 10.1186/s13075-023-03203-7

Figure Lengend Snippet: Fig. 1 Expression of Bcl-2 family members in CD19+ B cell subsets ex vivo and after activation on day 6. Immunological detection of Bcl-2, Bcl-XL, and Mcl-1 expression in CD19+ naïve (N; CD27-/CD38-), memory B cells (Mem; CD27+/CD38-), and plasmablasts (PB; CD27+/CD38+) of healthy controls (HD; N = 13) compared to rheumatoid arthritis (RA; N = 12), and systemic lupus erythematosus (SLE; N = 14) ex vivo (A–C) and after stimulation with 1 µg/ml ODN2006 (CpG) for 6 days (D–F). Filled symbols represent active disease, open symbols remission. Data are expressed as mean ± SD

Article Snippet: ODN2006 CpG oligonucleotide type B was purchased from InvivoGen (San Diego, CA, USA), venetoclax was purchased from Active Biochem (Bonn, Germany), S-63845 was purchased from Chemgood (Glen Allen, VA, USA), and AZD4320 was provided by Acerta Pharma (Oss, The Netherlands).

Techniques: Expressing, Ex Vivo, Activation Assay

Journal: Arthritis research & therapy

Article Title: Differential expression pattern of Bcl-2 family members in B and T cells in systemic lupus erythematosus and rheumatoid arthritis.

doi: 10.1186/s13075-023-03203-7

Figure Lengend Snippet: Fig. 2 CD19+ B cell proliferation following stimulation for 6 days. Cell proliferation was assessed by a CellTrace violet (CTV) staining in CD19+ B cells of healthy controls (HD) compared to RA/SLE patients after stimulation for 6 days with 1 µg/ml ODN2006 (CpG) or with T cell-dependent stimulation with CD3 (clone 1XE) and CD28 (clone 15E8) soluble antibodies. RA patients were all treated with methotrexate, while one patient also received prednisolone 5 mg/day (+). Two SLE patients were de novo (no mediation; 1+5), while most SLE patients received hydroxychloroquine (HCQ; 2,3,4) or a combination of HCQ with either azathioprine (6) or prednisone 15 mg/day (8), one SLE patient was treated with mycophenolate mofetil (7). Filled symbols represent active disease, open symbols remission. Two-way Anova test was used for statistical analyses; ns = not significant. Data are expressed as mean ± SD.

Article Snippet: ODN2006 CpG oligonucleotide type B was purchased from InvivoGen (San Diego, CA, USA), venetoclax was purchased from Active Biochem (Bonn, Germany), S-63845 was purchased from Chemgood (Glen Allen, VA, USA), and AZD4320 was provided by Acerta Pharma (Oss, The Netherlands).

Techniques: Staining

Journal: Arthritis research & therapy

Article Title: Differential expression pattern of Bcl-2 family members in B and T cells in systemic lupus erythematosus and rheumatoid arthritis.

doi: 10.1186/s13075-023-03203-7

Figure Lengend Snippet: Figure 3. BH3 mimetics revealed that Mcl-1 inhibition was the most effective in reducing plasmablast viability after activation. PBMCs of healthy controls (HD; N = 3), rheumatoid arthritis (RA; N = 3), and systemic lupus erythematosus (SLE; N = 4) were stimulated with 1 µg/ml ODN2006 (CpG) for 6 days followed by in vitro treatment with the Bcl-2 inhibitor venetoclax (A–C), Mcl-1 inhibitor S63845 (D–F), or dual Bcl-2/Bcl-XL inhibitor AZD4320 (G–I) for 24 h. Viability data were measured by flow cytometry using DiOC6/TO-PRO-3 staining in CD19+ naïve (N; CD27-/CD38-), memory B cells (Mem; CD27+/CD38-), and plasmablasts (PB; CD27+/CD38+). Two-way Anova test was used for statistical analyses. *p < 0.05; **p < 0.01; ***p < 0.001. Data are expressed as mean ± SEM

Article Snippet: ODN2006 CpG oligonucleotide type B was purchased from InvivoGen (San Diego, CA, USA), venetoclax was purchased from Active Biochem (Bonn, Germany), S-63845 was purchased from Chemgood (Glen Allen, VA, USA), and AZD4320 was provided by Acerta Pharma (Oss, The Netherlands).

Techniques: Inhibition, Activation Assay, In Vitro, Flow Cytometry, Staining

Journal: medRxiv

Article Title: IFNβ-1b treatment leads to changes in the B cell subset and cytokine secretion profile in patients with relapsing-remitting multiple sclerosis

doi: 10.1101/2022.02.25.22270266

Figure Lengend Snippet: Cryopreserved and thawed PBMC were ml in flat-bottom plates in AIM V medium with 10% human AB serum for 18 hours with cultured at 5×10 6 / 3 µg/ml CpG-ODN type B-2006-G5 at 37 ° C, 5% CO 2 . PMA (50 ng/ml) and ionomycin (750 ng/ml) were added after 12 hours. Brefeldin and monensin (PTI)(3 µg/ml and 2 µM, respectively) were added after 14 hours. PBMCs were stained for viability (yellow amine dye) prior to surface epitope and intracellular cytokine staining. Cytokine quadrants were set based on isotype controls.

Article Snippet: Cryopreserved and thawed PBMC (for both patients and healthy controls) were cultured at 5×10 6 /ml in 12-well, flat-bottom plates in AIM V medium supplemented with 10% pooled human AB serum for 18 hours with 3 µg/ml CpG oligodeoxynucleotide type B-2006-G5 (InvivoGen, CA) at 37 ° C in a humidified atmosphere containing 5% CO 2 .

Techniques: Cell Culture, Staining