cd20 6  (Servicebio Inc)

Journal: EMBO Reports

Article Title: T cell‐dependent bispecific antibodies alter organ‐specific endothelial cell–T cell interaction

doi: 10.15252/embr.202255532

Figure Lengend Snippet:

Article Snippet: PBMCs were treated with 0.5 μg/ml anti‐CD20/CD3 TDB (2H7v/6/38E4.v.1) and cultured over night at 37°C, 5% CO 2 in endothelial cell growth medium‐2 (EGMTM‐2 Endothelial Cell Growth Medium‐2 BulletKitTM, ref: CC‐3162, LONZA).

Techniques: Recombinant, Purification, Blocking Assay, Produced, RNAscope, Sequencing, Negative Control, Protease Inhibitor, Software, Multiplex Assay, Cell Isolation, Enzyme-linked Immunosorbent Assay, Library Quantification, BIA-KA, Gene Expression

Journal: Stem Cell Research & Therapy

Article Title: Construction of tandem diabody (IL-6/CD20)-secreting human umbilical cord mesenchymal stem cells and its experimental treatment on diffuse large B cell lymphoma

doi: 10.1186/s13287-022-03169-4

Figure Lengend Snippet: Construction of UCMSCs-Tandab(IL-6/CD20). A Schematic diagram of the Tandab recombinant vector (Tandab(CD20/IL-6)-6 × His-GFP vector). B Schematic representation to produce UCMSCs-Tandab(IL-6/CD20) and UCMSCs-Vector cell lines. With a lentivirus package, the Empty-GFP vector and Tandab(CD20/IL-6)-6 × His-GFP vector transfected HEK293T cells to collect empty and Tandab virus. Then, the empty and Tandab virus infected UCMSCs to produce UCMSCs-Vector and UCMSCs-Tandab(IL-6/CD20), respectively. C The morphology and fluorescence image of UCMSCs transfected with Tandab virus or empty virus under an inverted fluorescence microscope (Scale bar: 100 μm). Abbreviations: Tandab: Tandem diabody, GFP: Green fluorescent protein, 6 × His: Hexa-histidine tag

Article Snippet: The Tandab virus was produced from the transient transfection of the HEK293T cells (Ubigene, China) with Tandab(CD20/IL-6)-6 × His-GFP vector together with pCMV-VSVG envelope vector and psPAX2 packaging vector (Ubigene, China).

Techniques: Recombinant, Plasmid Preparation, Transfection, Virus, Infection, Fluorescence, Microscopy

Journal: Stem Cell Research & Therapy

Article Title: Construction of tandem diabody (IL-6/CD20)-secreting human umbilical cord mesenchymal stem cells and its experimental treatment on diffuse large B cell lymphoma

doi: 10.1186/s13287-022-03169-4

Figure Lengend Snippet: Vitality, apoptosis and homing abilities of modified UCMSCs. A Cell viability of UCMSCs-Tandab(IL-6/CD20) and UCMSCs-Vector by CCK-8 assay. n = 5 for each group. B Apoptosis assay of UCMSCs-Tandab(IL-6/CD20), UCMSCs-Vector and UCMSCs by flow cytometry analysis. The histogram of apoptosis is shown in ( C ). ( D ) The migration ability of all three UCMSCs lines by transwell migration assay. Supernatants of SU-DHL-4 were used to trigger cells (UCMSCs-Tandab(IL-6/CD20), UCMSCs-Vector and UCMSCs) migration, and the CFM group served as a negative control. The numbers of migrated cells were statistically analyzed in ( E ). n = 3 for each group. ns: not significant, * p < 0.05, ** p < 0.01. Abbreviations: CFM: Cell-free medium

Article Snippet: The Tandab virus was produced from the transient transfection of the HEK293T cells (Ubigene, China) with Tandab(CD20/IL-6)-6 × His-GFP vector together with pCMV-VSVG envelope vector and psPAX2 packaging vector (Ubigene, China).

Techniques: Modification, Plasmid Preparation, CCK-8 Assay, Apoptosis Assay, Flow Cytometry, Migration, Transwell Migration Assay, Negative Control

Journal: Stem Cell Research & Therapy

Article Title: Construction of tandem diabody (IL-6/CD20)-secreting human umbilical cord mesenchymal stem cells and its experimental treatment on diffuse large B cell lymphoma

doi: 10.1186/s13287-022-03169-4

Figure Lengend Snippet: The expression of Tandab protein. ( A ) The relative expression of Tandab mRNA in UCMSCs-Tandab(IL-6/CD20), UCMSCs-Vector and UCMSCs analyzed by qRT-PCR. ( B-C ) The relative expression of Tandab protein in the cell lysis ( B ) and the culture supernatant ( C ) of UCMSCs-Tandab(IL-6/CD20) by western blot. ( D ) Representative immunofluorescence staining of UCMSCs-Tandab(IL-6/CD20), UCMSCs-Vector and UCMSCs. Green and red represent spontaneous fluorescence and 6 × His Tag, respectively. ( E–F ) After collecting the culture supernatants of UCMSCs-Tandab(IL-6/CD20), the Tandab protein level was measured by ELISA. n = 3 for each group. ns: not significant, *** p < 0.001

Article Snippet: The Tandab virus was produced from the transient transfection of the HEK293T cells (Ubigene, China) with Tandab(CD20/IL-6)-6 × His-GFP vector together with pCMV-VSVG envelope vector and psPAX2 packaging vector (Ubigene, China).

Techniques: Expressing, Plasmid Preparation, Quantitative RT-PCR, Lysis, Western Blot, Immunofluorescence, Staining, Fluorescence, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Research & Therapy

Article Title: Construction of tandem diabody (IL-6/CD20)-secreting human umbilical cord mesenchymal stem cells and its experimental treatment on diffuse large B cell lymphoma

doi: 10.1186/s13287-022-03169-4

Figure Lengend Snippet: Tandab bind to the CD20-positive SU-DHL-2/4 cells. A The fluorescence image of Tandab bound to SU-DHL-2 and SU-DHL-4 cells. Green and red represent CD20 and 6 × His Tag, respectively. B Flow cytometric analysis of Tandab bound to SU-DHL-2/4 cells. Blue: negative control, red: experimental groups treated with UCMSCs-Tandab(IL-6/CD20). C The competitive binding assay with anti-human CD20 monoclonal antibody. Blue: negative control, red: Tandab(IL-6/CD20) + anti-human CD20 monoclonal antibody, orange: anti-human CD20 monoclonal antibody alone

Article Snippet: The Tandab virus was produced from the transient transfection of the HEK293T cells (Ubigene, China) with Tandab(CD20/IL-6)-6 × His-GFP vector together with pCMV-VSVG envelope vector and psPAX2 packaging vector (Ubigene, China).

Techniques: Fluorescence, Negative Control, Competitive Binding Assay

Journal: Stem Cell Research & Therapy

Article Title: Construction of tandem diabody (IL-6/CD20)-secreting human umbilical cord mesenchymal stem cells and its experimental treatment on diffuse large B cell lymphoma

doi: 10.1186/s13287-022-03169-4

Figure Lengend Snippet: UCMSCs-Tandab(IL-6/CD20) inhibited the proliferation of SU-DHL-2/4. ( A-D ) CCK-8 assay for evaluating the cell viability of SU-DHL-2 ( A and C ) or SU-DHL-4 ( B and D ) after different treatments (the supernatants of UCMSCs-Tandab(IL-6/CD20), Rituximab (1000 ng/ml, 100 ng/ml, 10 ng/ml), Tocilizumab(10 ng/ml) and mixture of Rituximab and Tocilizumab (5 and 5 ng/ml)) without the presence of PBMCs. ( E–H ) CCK-8 assay for evaluating the viability of SU-DHL-2 ( E and G ) or SU-DHL-4 ( F and H ) after different treatments (the supernatants of UCMSCs-Tandab(IL-6/CD20), Rituximab(1000 ng/ml, 100 ng/ml, 10 ng/ml), Tocilizumab(10 ng/ml) and mixture of Rituximab and Tocilizumab (5 and 5 ng/ml)) with the presence of PBMCs. n = 6 for each group. ns: not significant, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The Tandab virus was produced from the transient transfection of the HEK293T cells (Ubigene, China) with Tandab(CD20/IL-6)-6 × His-GFP vector together with pCMV-VSVG envelope vector and psPAX2 packaging vector (Ubigene, China).

Techniques: CCK-8 Assay

Journal: Stem Cell Research & Therapy

Article Title: Construction of tandem diabody (IL-6/CD20)-secreting human umbilical cord mesenchymal stem cells and its experimental treatment on diffuse large B cell lymphoma

doi: 10.1186/s13287-022-03169-4

Figure Lengend Snippet: UCMSCs-Tandab(IL-6/CD20) inhibited the proliferation of SU-DHL-2/4. ( A-B ) CCK-8 assay to detect the toxicity effective of different E/T values (0:1, 0.1:1, 1:1, 10:1) on SU-DHL-2 (A) or SU-DHL-4 ( B ) cells. The solid lines indicate the groups treated with the supernatants of UCMSCs-Tandab(IL-6/CD20). n = 6 for each group. ( C-F ) EDU assay to evaluate the cell viability of SU-DHL-2 or SU-DHL-4 after different treatments (UCMSCs-Tandab(IL-6/CD20), UCMSCs-Vector, UCMSCs, Rituximab, Tocilizumab, mixture of Rituximab and Tocilizumab) with ( E–F ) or without ( C-D ) the presence of PBMCs and analytical results of positively stained EDU-positive cells to detect the percentage of proliferating cells after cell cultured for 72 h in ( D ) and ( F ). n = 5 for each group. ns: not significant, * p < 0.05, ** p < 0.01

Article Snippet: The Tandab virus was produced from the transient transfection of the HEK293T cells (Ubigene, China) with Tandab(CD20/IL-6)-6 × His-GFP vector together with pCMV-VSVG envelope vector and psPAX2 packaging vector (Ubigene, China).

Techniques: CCK-8 Assay, EdU Assay, Plasmid Preparation, Staining, Cell Culture

Journal: Frontiers in Immunology

Article Title: Distribution and efficacy of ofatumumab and ocrelizumab in humanized CD20 mice following subcutaneous or intravenous administration

doi: 10.3389/fimmu.2022.814064

Figure Lengend Snippet: (A) Representative maximum intensity projection (MIP) SPECT/CT images of huCD20 (upper panel) and WT (lower panel) mice 72 h after SC or IV administration of 111 In-ofatumumab or 111 In-ocrelizumab. Prominent binding of 111 In-ofatumumab and 111 In-ocrelizumab is observed in lymph nodes of huCD20 mice following SC administration, but not after IV administration. Both 111 In-ofatumumab and 111 In-ocrelizumab displayed similar tissue distributions. No specific binding was observed in WT mice. B = bladder, C = clavicle, H = heart, K = kidney, L = liver, LN = lymph node, Sp = spleen (B) VOI analysis of SPECT/CT imaging (N = 5 per group). A significantly higher level of binding was observed in axillary and inguinal lymph nodes of huCD20 mice following SC versus IV administration. * p < 0.05; ** p < 0.01. Data represent mean ± SEM.

Article Snippet: Adult huCD20 C57BL/6 mice (B6.FVB.Tg[CD20]Gne), expressing human CD20 exclusively on B-cells , and their wildtype (WT) littermates (JANVIER LABS, France) were housed in standard laboratory cages (3–5 per cage) in a controlled enriched environment with a 12:12 h light–dark cycle, and with food and water available ad libitum .

Techniques: Single Photon Emission Computed Tomography, Binding Assay, Imaging

Journal: Frontiers in Immunology

Article Title: Distribution and efficacy of ofatumumab and ocrelizumab in humanized CD20 mice following subcutaneous or intravenous administration

doi: 10.3389/fimmu.2022.814064

Figure Lengend Snippet: Gamma counting analysis of 111 In-ofatumumab and 111 In-ocrelizumab in the blood and individual organs of WT and huCD20 mice. (A) Non-specific accumulation of 111 In-ofatumumab and 111 In-ocrelizumab was observed in the tissue of WT mice. (B) Increased levels of 111 In-ocrelizumab and 111 In-ofatumumab were observed in the B-cell rich spleen of huCD20 mice. 111 In-ofatumumab-treated huCD20 mice displayed faster clearance from the blood and uptake in the spleen following both SC and IV administration compared with 111 In-ocrelizumab. * p < 0.05; ** p < 0.01; *** p < 0.001. Data represent mean ± SEM.

Article Snippet: Adult huCD20 C57BL/6 mice (B6.FVB.Tg[CD20]Gne), expressing human CD20 exclusively on B-cells , and their wildtype (WT) littermates (JANVIER LABS, France) were housed in standard laboratory cages (3–5 per cage) in a controlled enriched environment with a 12:12 h light–dark cycle, and with food and water available ad libitum .

Techniques:

Journal: Frontiers in Immunology

Article Title: Distribution and efficacy of ofatumumab and ocrelizumab in humanized CD20 mice following subcutaneous or intravenous administration

doi: 10.3389/fimmu.2022.814064

Figure Lengend Snippet: (A) Representative MIP SPECT/CT images of huCD20 and WT mice 72 h after SC or IV administration of 111 In-anti-mouse CD19. Prominent binding of 111 In-anti-CD19 is observed in lymph nodes of huCD20 and WT mice following SC administration, but not after IV administration. J = joint; L = liver, LN = lymph node, Sp = spleen. (B) Quantitative VOI analysis of SPECT/CT imaging. (C) Uptake of 111 In-anti-CD19 in the blood and tissue of WT mice 72 h after administration, as assessed by ex-vivo gamma counting analysis. LF = right lower flank, NN = nape of the neck. * p < 0.05; ** p < 0.01. Data represent mean ± SEM.

Article Snippet: Adult huCD20 C57BL/6 mice (B6.FVB.Tg[CD20]Gne), expressing human CD20 exclusively on B-cells , and their wildtype (WT) littermates (JANVIER LABS, France) were housed in standard laboratory cages (3–5 per cage) in a controlled enriched environment with a 12:12 h light–dark cycle, and with food and water available ad libitum .

Techniques: Single Photon Emission Computed Tomography, Binding Assay, Imaging, Ex Vivo

Journal: Frontiers in Immunology

Article Title: Distribution and efficacy of ofatumumab and ocrelizumab in humanized CD20 mice following subcutaneous or intravenous administration

doi: 10.3389/fimmu.2022.814064

Figure Lengend Snippet: (A) Representative MIP SPECT/CT images of huCD20 mice 72 h after SC administration of 111 In-anti-CD19 (pre) and 72 h after SC administration of 111 In-anti-CD19 following 18 days of treatment with SC or IV ofatumumab or ocrelizumab (post). A reduction in 111 In-anti-CD19 signal in lymph nodes, representing B-cell depletion, is noted post treatment. (B) Two-way ANOVA revealed that all treatments significantly reduced the 111 In-anti-CD19 signal over time in the cervical lymph nodes and axillary lymph nodes, but no significant difference in SUVmax was observed between groups (route of administration or antibody). There was also no significant difference in the relative decrease in SUVmax between groups, as assessed by one-way ANOVA. Data represent mean ± SEM.

Article Snippet: Adult huCD20 C57BL/6 mice (B6.FVB.Tg[CD20]Gne), expressing human CD20 exclusively on B-cells , and their wildtype (WT) littermates (JANVIER LABS, France) were housed in standard laboratory cages (3–5 per cage) in a controlled enriched environment with a 12:12 h light–dark cycle, and with food and water available ad libitum .

Techniques: Single Photon Emission Computed Tomography