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ACROBiosystems human cd79b protein
Human Cd79b Protein, supplied by ACROBiosystems, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+cd79b+protein/Human+CD79B+Protein%2C+Fc+Tag/pm41814478-143-15-19
Average 95 stars, based on 1 article reviews

human cd79b protein - by Bioz Stars, 2026-10

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Article Title: Direct Visualization and Quantitative Modeling of Mass Transport Limitation in Biomolecular Kinetics via Imaging-Based Meta-SPR.
Article Snippet: Directly observing how mass transport limitation (MTL) distorts biomolecular kinetics has long remained out of reach.. Here, we harness an imaging-based metasurface plasmon resonance (Meta-SPR) platform to both visualize and chemically quantify MTL effects in real time, revealing a pronounced flow-path signal decay beyond bulk depletion.. This spatially resolved capability uniquely links optical contrast to the local analyte concentration, enabling quantitative mapping of MTL severity.

Image Search Results

Concept of novel one-step and site-specific MTG technology and manufacturing of anti–CD79b-RKAA-MMAE. A, Schematic drawing of the novel MTG-based conjugation technology using a linker-payload composed of a small RKAA peptide linker and the cytotoxic payload MMAE. Functionalization occurs at the Q295 residue of the native, glycosylated polatuzumab antibody. RKAA-MMAE is attached in a single step using wild-type MTG. B, Linker-payload structure comprised the RKAA-peptide linker with an acetylated N-terminus, a PABC self-immolative spacer, and MMAE. C, Schematic comparison of anti–CD79b-RKAA-MMAE generated by one-step site-specific MTG conjugation and FDA-approved PV.

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Concept of novel one-step and site-specific MTG technology and manufacturing of anti–CD79b-RKAA-MMAE. A, Schematic drawing of the novel MTG-based conjugation technology using a linker-payload composed of a small RKAA peptide linker and the cytotoxic payload MMAE. Functionalization occurs at the Q295 residue of the native, glycosylated polatuzumab antibody. RKAA-MMAE is attached in a single step using wild-type MTG. B, Linker-payload structure comprised the RKAA-peptide linker with an acetylated N-terminus, a PABC self-immolative spacer, and MMAE. C, Schematic comparison of anti–CD79b-RKAA-MMAE generated by one-step site-specific MTG conjugation and FDA-approved PV.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Conjugation Assay, Residue, Comparison, Generated

Analytic characterization of anti–CD79b-RKAA-MMAE. A, LC/MS spectra of the heavy chain (HC) and light chain (LC) of glycosylated anti–CD79b-RKAA-MMAE under reducing conditions. Main peaks showing single conjugation of HC (left graph) and unconjugated LC (right graph). B, Intact LC/MS. Main peaks showing DAR 2 ADC. C, Reverse-phase UHPLC analysis of reduced anti–CD79b-RKAA-MMAE. D, Analysis of monomer content by SEC. E, Comparison of HIC profiles of unconjugated polatuzumab antibody, anti–CD79b-RKAA-MMAE, and PV. Different DAR species of the preparations are indicated.

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Analytic characterization of anti–CD79b-RKAA-MMAE. A, LC/MS spectra of the heavy chain (HC) and light chain (LC) of glycosylated anti–CD79b-RKAA-MMAE under reducing conditions. Main peaks showing single conjugation of HC (left graph) and unconjugated LC (right graph). B, Intact LC/MS. Main peaks showing DAR 2 ADC. C, Reverse-phase UHPLC analysis of reduced anti–CD79b-RKAA-MMAE. D, Analysis of monomer content by SEC. E, Comparison of HIC profiles of unconjugated polatuzumab antibody, anti–CD79b-RKAA-MMAE, and PV. Different DAR species of the preparations are indicated.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Liquid Chromatography with Mass Spectroscopy, Conjugation Assay, Comparison

In vitro evaluation of binding, toxicity, and stability of anti–CD79b-RKAA-MMAE. A, Binding comparison of unconjugated polatuzumab, anti–CD79b-RKAA-MMAE, and PV on recombinant CD79b by ELISA. The graph shows mean ± SD of technical duplicates. B, Representative assay to evaluate target binding on CD79b-positive BJAB cells. C, Evaluation of Fc functionality by ELISA on immobilized FcγRI (CD64). D, In vitro cytotoxicity assay on CD79b-expressing Granta-519 cells. Comparison of anti–CD79b-RKAA-MMAE (DAR 2) and PV (DAR 3.5) at equal ADC concentrations. E, Toxicity on CD79b-positive WSU-DLCL2 tumor cells. F, In vitro cytotoxicity assay on target-negative HT cells. G, All cytotoxicity graphs show mean ± SD of technical duplicates. A representative serum stability assay of anti–CD79b-RKAA-MMAE in human, cynomolgus monkey, and mouse IgG-depleted sera at 37°C for 7 days. DAR was assessed by LC/MS at different time points. H, Evaluation of PV stability in human, cynomolgus monkey, and mouse IgG-depleted sera. MFI, mean fluorescence intensity; OD, optical density.

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: In vitro evaluation of binding, toxicity, and stability of anti–CD79b-RKAA-MMAE. A, Binding comparison of unconjugated polatuzumab, anti–CD79b-RKAA-MMAE, and PV on recombinant CD79b by ELISA. The graph shows mean ± SD of technical duplicates. B, Representative assay to evaluate target binding on CD79b-positive BJAB cells. C, Evaluation of Fc functionality by ELISA on immobilized FcγRI (CD64). D, In vitro cytotoxicity assay on CD79b-expressing Granta-519 cells. Comparison of anti–CD79b-RKAA-MMAE (DAR 2) and PV (DAR 3.5) at equal ADC concentrations. E, Toxicity on CD79b-positive WSU-DLCL2 tumor cells. F, In vitro cytotoxicity assay on target-negative HT cells. G, All cytotoxicity graphs show mean ± SD of technical duplicates. A representative serum stability assay of anti–CD79b-RKAA-MMAE in human, cynomolgus monkey, and mouse IgG-depleted sera at 37°C for 7 days. DAR was assessed by LC/MS at different time points. H, Evaluation of PV stability in human, cynomolgus monkey, and mouse IgG-depleted sera. MFI, mean fluorescence intensity; OD, optical density.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: In Vitro, Binding Assay, Comparison, Recombinant, Enzyme-linked Immunosorbent Assay, Cytotoxicity Assay, Expressing, Stability Assay, Liquid Chromatography with Mass Spectroscopy, Fluorescence

Enzymatic cleavage of anti–CD79b-RKAA-MMAE. Comparison of the cleavage rate of anti–CD79b-RKAA-MMAE and PV using human cathepsin B ( A ), mouse Ces1c ( B ), or HLL extract ( C ). D, Schematic overview of the cleavage mechanism of anti–CD79b-RKAA-MMAE by proteases and subsequent payload release and liberation of free MMAE by 1,6-elimination of PABC.

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Enzymatic cleavage of anti–CD79b-RKAA-MMAE. Comparison of the cleavage rate of anti–CD79b-RKAA-MMAE and PV using human cathepsin B ( A ), mouse Ces1c ( B ), or HLL extract ( C ). D, Schematic overview of the cleavage mechanism of anti–CD79b-RKAA-MMAE by proteases and subsequent payload release and liberation of free MMAE by 1,6-elimination of PABC.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Comparison

Pharmacokinetic studies in Swiss wild-type mice and Sprague Dawley rats. A, Healthy female Swiss wild-type mice were injected intravenously with 5 mg/kg unconjugated polatuzumab, anti–CD79b-RKAA-MMAE, or PV ( n = 5 per group). Plasma was collected at different time points after injection, and total IgG and intact ADC concentrations were determined by ELISA. Data represent mean plasma concentrations (±SD). B, Male Sprague Dawley rats were injected intravenously with 10 mg/kg of anti–CD79b-RKAA-MMAE (top graph) or 10 mg/kg of PV (bottom graph) in a repeat-dose study ( n = 5 per group). Animals were injected weekly for a total of four injections. Graphs show plasma concentration–time curves after the fourth dose, including the predose concentration measurement (0 hours).

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Pharmacokinetic studies in Swiss wild-type mice and Sprague Dawley rats. A, Healthy female Swiss wild-type mice were injected intravenously with 5 mg/kg unconjugated polatuzumab, anti–CD79b-RKAA-MMAE, or PV ( n = 5 per group). Plasma was collected at different time points after injection, and total IgG and intact ADC concentrations were determined by ELISA. Data represent mean plasma concentrations (±SD). B, Male Sprague Dawley rats were injected intravenously with 10 mg/kg of anti–CD79b-RKAA-MMAE (top graph) or 10 mg/kg of PV (bottom graph) in a repeat-dose study ( n = 5 per group). Animals were injected weekly for a total of four injections. Graphs show plasma concentration–time curves after the fourth dose, including the predose concentration measurement (0 hours).

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Injection, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Concentration Assay

Therapeutic efficacy of anti–CD79b-RKAA-MMAE in different xenograft models. Treatment was started when tumors reached a size of approximately 200 mm 3 . Mice were allocated into different groups using a nonrandom stratification protocol. Data represent mean tumor volumes (±SEM). A, CB17-SCID mice were challenged with 20 × 10 6 Granta-519 tumor cells and received a single injection (black arrow) of 2.1 mg/kg anti–CD79b-RKAA-MMAE (corresponding to 20 μg/kg of payload), 2.1 mg/kg PV (corresponding to 40 μg/kg of payload), or PBS, intravenously into the lateral tail vein ( n = 8 per group). B, Treatment of the Granta-519 xenograft model at equal payload doses of 10 µg/kg, corresponding to 1 mg/kg of anti–CD79b-RKAA-MMAE or 0.53 mg/kg of PV ( n = 8 per group). C, Administration of 1.5 mg/kg (15 μg/kg of payload) of anti–CD79b-RKAA-MMAE or nonbinding control IgG1-RKAA-MMAE to Granta-519–bearing CB17-SCID mice. D, CB17-SCID mice were challenged with 20 × 10 6 Ramos Burkitt lymphoma cells. Anti–CD79b-RKAA-MMAE (2.5 mg/kg) or PV (1.43 mg/kg; both corresponding to 25 μg/kg of payload) was used for treatment ( n = 6 per group). PBS was used as a negative control. *, P < 0.05; ****, P < 0.0001. CR, complete response.

Journal: Molecular Cancer Therapeutics

Article Title: Broadening the Therapeutic Window of ADCs Using Site-Specific Bioconjugation Showcased by an MMAE-Containing Peptide Linker in a CD79b-Targeting ADC

doi: 10.1158/1535-7163.MCT-24-0983

Figure Lengend Snippet: Therapeutic efficacy of anti–CD79b-RKAA-MMAE in different xenograft models. Treatment was started when tumors reached a size of approximately 200 mm 3 . Mice were allocated into different groups using a nonrandom stratification protocol. Data represent mean tumor volumes (±SEM). A, CB17-SCID mice were challenged with 20 × 10 6 Granta-519 tumor cells and received a single injection (black arrow) of 2.1 mg/kg anti–CD79b-RKAA-MMAE (corresponding to 20 μg/kg of payload), 2.1 mg/kg PV (corresponding to 40 μg/kg of payload), or PBS, intravenously into the lateral tail vein ( n = 8 per group). B, Treatment of the Granta-519 xenograft model at equal payload doses of 10 µg/kg, corresponding to 1 mg/kg of anti–CD79b-RKAA-MMAE or 0.53 mg/kg of PV ( n = 8 per group). C, Administration of 1.5 mg/kg (15 μg/kg of payload) of anti–CD79b-RKAA-MMAE or nonbinding control IgG1-RKAA-MMAE to Granta-519–bearing CB17-SCID mice. D, CB17-SCID mice were challenged with 20 × 10 6 Ramos Burkitt lymphoma cells. Anti–CD79b-RKAA-MMAE (2.5 mg/kg) or PV (1.43 mg/kg; both corresponding to 25 μg/kg of payload) was used for treatment ( n = 6 per group). PBS was used as a negative control. *, P < 0.05; ****, P < 0.0001. CR, complete response.

Article Snippet: Plasma concentrations were determined by ELISA using His-tagged human CD79b (Sino Biological) as the capturing agent, coated on Ni-NTA HisSorb plates (Qiagen).

Techniques: Drug discovery, Injection, Control, Negative Control

Figure 1 Schematic of different CD79b CAR constructs and transduction efficiency in primary T cells. (A) Schematic design of CAR constructs to target CD19 or CD79b. (B and C) CAR constructs shown in panel A were transduced lentivirally into primary human T cells and transduction efficiency was determined by flow cytometry by assessing eGFP expression and CAR expression is shown by demonstrating binding to fluorescently labeled recombinant human CD79b-Fc protein. Data is representative of CAR T cells generated from two different donors. CAR, chimeric antigen receptor; eGFP, enhanced green fluorescent protein; LTR, long terminal repeat; LTRtr, LTR truncated; scFv, single chain variable fragment; sp, signal peptide; VL, variable light chain; VH, variable heavy chain; TM, transmembrane domain.

Journal: Journal for immunotherapy of cancer

Article Title: Chimeric antigen receptor T cells to target CD79b in B-cell lymphomas.

doi: 10.1136/jitc-2023-007515

Figure Lengend Snippet: Figure 1 Schematic of different CD79b CAR constructs and transduction efficiency in primary T cells. (A) Schematic design of CAR constructs to target CD19 or CD79b. (B and C) CAR constructs shown in panel A were transduced lentivirally into primary human T cells and transduction efficiency was determined by flow cytometry by assessing eGFP expression and CAR expression is shown by demonstrating binding to fluorescently labeled recombinant human CD79b-Fc protein. Data is representative of CAR T cells generated from two different donors. CAR, chimeric antigen receptor; eGFP, enhanced green fluorescent protein; LTR, long terminal repeat; LTRtr, LTR truncated; scFv, single chain variable fragment; sp, signal peptide; VL, variable light chain; VH, variable heavy chain; TM, transmembrane domain.

Article Snippet: Surface expression of CD79b CAR was detected using fluorochrome labeled recombinant human CD79b- Fc protein (Creative BioMart) or goat anti- mouse IgG F(ab)2 fragment (Jackson ImmunoResearch).

Techniques: Construct, Transduction, Flow Cytometry, Expressing, Binding Assay, Labeling, Recombinant, Generated

Figure 2 CD79b CAR T cells proliferated and produced cytokines in response to lymphoma cells in vitro. CAR T cells targeting CD19 or CD79b generated from healthy donor T cells were co-cultured with Daudi Burkitt lymphoma tumor cells at an effector:target ratio of 1:1. Untransduced T cells were used as controls. (A) Representative flow cytometric plots illustrating proliferation of CD4+ and CD8+ CAR T cells after 96 hours of co-culture. (B) Summary results showing proliferation from replicate wells (N=2). (C) Cytokine production by CAR T cells in response to Daudi lymphoma cells in replicate wells (N=2) after 24 hours of co-culture. Data represents mean±SD and is representative of at least three separate experiments from three different donors. ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05. CAR, chimeric antigen receptor; IFN, interferon; IL, interleukin; GM- CSF, granulocyte-macrophage colony-stimulating factor.

Journal: Journal for immunotherapy of cancer

Article Title: Chimeric antigen receptor T cells to target CD79b in B-cell lymphomas.

doi: 10.1136/jitc-2023-007515

Figure Lengend Snippet: Figure 2 CD79b CAR T cells proliferated and produced cytokines in response to lymphoma cells in vitro. CAR T cells targeting CD19 or CD79b generated from healthy donor T cells were co-cultured with Daudi Burkitt lymphoma tumor cells at an effector:target ratio of 1:1. Untransduced T cells were used as controls. (A) Representative flow cytometric plots illustrating proliferation of CD4+ and CD8+ CAR T cells after 96 hours of co-culture. (B) Summary results showing proliferation from replicate wells (N=2). (C) Cytokine production by CAR T cells in response to Daudi lymphoma cells in replicate wells (N=2) after 24 hours of co-culture. Data represents mean±SD and is representative of at least three separate experiments from three different donors. ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05. CAR, chimeric antigen receptor; IFN, interferon; IL, interleukin; GM- CSF, granulocyte-macrophage colony-stimulating factor.

Techniques: Produced, In Vitro, Generated, Cell Culture, Co-Culture Assay

Figure 3 CD79b CAR T cells degranulated and exhibited cytotoxic activity against lymphoma cells in vitro. CAR T cells targeting CD19 or CD79b were generated from healthy donor T cells and co-cultured with Daudi Burkitt lymphoma tumor cells at an effector:target ratio of 1:1. Untransduced T cells were used as controls. (A) Representative flow cytometric plots illustrating CD107a/b degranulation on CD4+ and CD8+ CAR T cells after 6 hours of co-culture are shown. (B) Summary results showing degranulation from replicate wells (N=2). (C–E) CAR T cells labeled with CellTrace Far Red were co-cultured with Daudi lymphoma tumor cells labeled with CellTrace Violet at an effector:target ratio of 1:1. Cytotoxicity assay was assessed after 96 hours by flow cytometry by counting live tumor cells using TruCOUNT beads. Untransduced T cells were used as controls. (C) Representative flow cytometric plots showing percentage of live tumor and T cells after end of co-culture. Summary results of cytotoxicity assay from replicate wells (N=2) showing absolute live tumor cell count (D) or specific lysis (E) of Daudi tumor cells. Data represents mean±SD and is representative of at least three separate experiments from three different donors. ***p<0.001; **p<0.01; *p<0.05; (ns)p>0.05. CAR, chimeric antigen receptor.

Journal: Journal for immunotherapy of cancer

Article Title: Chimeric antigen receptor T cells to target CD79b in B-cell lymphomas.

doi: 10.1136/jitc-2023-007515

Figure Lengend Snippet: Figure 3 CD79b CAR T cells degranulated and exhibited cytotoxic activity against lymphoma cells in vitro. CAR T cells targeting CD19 or CD79b were generated from healthy donor T cells and co-cultured with Daudi Burkitt lymphoma tumor cells at an effector:target ratio of 1:1. Untransduced T cells were used as controls. (A) Representative flow cytometric plots illustrating CD107a/b degranulation on CD4+ and CD8+ CAR T cells after 6 hours of co-culture are shown. (B) Summary results showing degranulation from replicate wells (N=2). (C–E) CAR T cells labeled with CellTrace Far Red were co-cultured with Daudi lymphoma tumor cells labeled with CellTrace Violet at an effector:target ratio of 1:1. Cytotoxicity assay was assessed after 96 hours by flow cytometry by counting live tumor cells using TruCOUNT beads. Untransduced T cells were used as controls. (C) Representative flow cytometric plots showing percentage of live tumor and T cells after end of co-culture. Summary results of cytotoxicity assay from replicate wells (N=2) showing absolute live tumor cell count (D) or specific lysis (E) of Daudi tumor cells. Data represents mean±SD and is representative of at least three separate experiments from three different donors. ***p<0.001; **p<0.01; *p<0.05; (ns)p>0.05. CAR, chimeric antigen receptor.

Techniques: Activity Assay, In Vitro, Generated, Cell Culture, Co-Culture Assay, Labeling, Cytotoxicity Assay, Flow Cytometry, Cell Counting, Lysis

Figure 4 CD79b CAR T cells exert antitumor effects in vivo. (A) Schematic overview of xenograft mouse model. Luciferase- labeled Daudi Burkitt lymphoma cells were injected intravenously into NSG mice at 2×104 tumor cells/mouse. After 11 days, mice were treated with untransduced T cells, CD19 CAR T cells, or CD79b CAR T cells via tail vein injection at 5×106 CAR+ T cells/mouse. (B) Tumor burden assessed by bioluminescence imaging at the indicated time points is shown. (C) Average radiance (photons/sec/cm2/steradian) of groups of mice at different time points is shown. Significance among treatment groups was determined by comparing area under the curve for each CAR-T group to untransduced control T-cell group in an ordinary one-way ANOVA with Dunnett’s multiple comparisons test (n=5 mice per group, mean±SD is shown). (D) Kaplan-Meier survival curves of groups of mice treated with different T cells. (****p<0.0001; ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05, ANOVA). ANOVA, analysis of variance; CAR, chimeric antigen receptor; I.V., intravenous.

Journal: Journal for immunotherapy of cancer

Article Title: Chimeric antigen receptor T cells to target CD79b in B-cell lymphomas.

doi: 10.1136/jitc-2023-007515

Figure Lengend Snippet: Figure 4 CD79b CAR T cells exert antitumor effects in vivo. (A) Schematic overview of xenograft mouse model. Luciferase- labeled Daudi Burkitt lymphoma cells were injected intravenously into NSG mice at 2×104 tumor cells/mouse. After 11 days, mice were treated with untransduced T cells, CD19 CAR T cells, or CD79b CAR T cells via tail vein injection at 5×106 CAR+ T cells/mouse. (B) Tumor burden assessed by bioluminescence imaging at the indicated time points is shown. (C) Average radiance (photons/sec/cm2/steradian) of groups of mice at different time points is shown. Significance among treatment groups was determined by comparing area under the curve for each CAR-T group to untransduced control T-cell group in an ordinary one-way ANOVA with Dunnett’s multiple comparisons test (n=5 mice per group, mean±SD is shown). (D) Kaplan-Meier survival curves of groups of mice treated with different T cells. (****p<0.0001; ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05, ANOVA). ANOVA, analysis of variance; CAR, chimeric antigen receptor; I.V., intravenous.

Techniques: In Vivo, Luciferase, Labeling, Injection, Imaging, Control

Figure 5 CD79b CAR T cells generated from patients with lymphoma relapsing after autologous CD19 CAR T-cell therapy are functional. CAR T cells targeting CD79b were generated from T cells isolated from two patients with large B-cell lymphoma relapsing after autologous CD19 CAR T-cell therapy and co-cultured with Daudi Burkitt lymphoma tumor cells at an effector:target ratio of 1:1. Untransduced T cells were used as controls. Representative data from one patient is shown. (A) Representative flow cytometric plots showing transduction efficiency of 79b CAR and proportion of CD4+ and CD8+ T cells. (B) Representative flow cytometric plots illustrating CD107a/b degranulation by CD4+ and CD8+ CAR T cells after 6 hours of co-culture. (C) Summary results showing degranulation from replicate wells (N=2). (D) Representative flow cytometric plots illustrating proliferation of CD4+ and CD8+ CAR T cells after 96 hours of co-culture. (E) Summary results showing proliferation from replicate wells (N=2). (F) Summary results of cytotoxicity assay from replicate wells (N=2) showing absolute live tumor cell count (left panel) or specific lysis (right panel). (G) Cytokines production by CAR T cells co-cultured with Daudi lymphoma cells in replicate wells (N=2) after 24 hours of co-culture. Data is representative of at least three separate experiments with T cells from two different patients. ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05. CAR, chimeric antigen receptor; IFN, interferon; IL, interleukin; GFP, green fluorescent protein; GM-CSF, granulocyte-macrophage colony-stimulating factor.

Journal: Journal for immunotherapy of cancer

Article Title: Chimeric antigen receptor T cells to target CD79b in B-cell lymphomas.

doi: 10.1136/jitc-2023-007515

Figure Lengend Snippet: Figure 5 CD79b CAR T cells generated from patients with lymphoma relapsing after autologous CD19 CAR T-cell therapy are functional. CAR T cells targeting CD79b were generated from T cells isolated from two patients with large B-cell lymphoma relapsing after autologous CD19 CAR T-cell therapy and co-cultured with Daudi Burkitt lymphoma tumor cells at an effector:target ratio of 1:1. Untransduced T cells were used as controls. Representative data from one patient is shown. (A) Representative flow cytometric plots showing transduction efficiency of 79b CAR and proportion of CD4+ and CD8+ T cells. (B) Representative flow cytometric plots illustrating CD107a/b degranulation by CD4+ and CD8+ CAR T cells after 6 hours of co-culture. (C) Summary results showing degranulation from replicate wells (N=2). (D) Representative flow cytometric plots illustrating proliferation of CD4+ and CD8+ CAR T cells after 96 hours of co-culture. (E) Summary results showing proliferation from replicate wells (N=2). (F) Summary results of cytotoxicity assay from replicate wells (N=2) showing absolute live tumor cell count (left panel) or specific lysis (right panel). (G) Cytokines production by CAR T cells co-cultured with Daudi lymphoma cells in replicate wells (N=2) after 24 hours of co-culture. Data is representative of at least three separate experiments with T cells from two different patients. ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05. CAR, chimeric antigen receptor; IFN, interferon; IL, interleukin; GFP, green fluorescent protein; GM-CSF, granulocyte-macrophage colony-stimulating factor.

Techniques: Generated, Functional Assay, Isolation, Cell Culture, Transduction, Co-Culture Assay, Cytotoxicity Assay, Cell Counting, Lysis

Figure 6 Efficacy of CD79b CAR T cells against patient-derived lymphoma tumor cells. CAR T cells targeting CD79b were generated from primary healthy donor T cells, labeled with CellTrace Far Red and co-cultured with Daudi Burkitt lymphoma tumor cells or lymphoma tumor cells derived from two PDXs (PDX203 and PDX300) for 96 hours. Untransduced T cells were used as controls. (A) Histograms showing expression of CD19 and CD79b in the two tumor samples. (B) Representative flow cytometric plots illustrating proliferation of CD4+ and CD8+ CARs T cells in response to tumor cells at an effector:target (E:T) ratio of 0.6:1. (C) Summary results showing proliferation from replicate wells (N=2). (D) Summary results of cytotoxicity assay from replicate wells (N=2) showing specific lysis at E:T ratio of 0.6:1. (E and F) Pooled data from 17 experiments testing CD79b CAR T cells generated from five donors is shown for per cent change in absolute number of live tumor cells on day 4 in an in vitro cytotoxicity assay compared with day 0 (E) and fold change in interferon-γ production compared with untransduced T cells after 24 hours of co-culture (F). Target tumor cells in these experiments included Daudi, Jeko-1, SUDHL6, or PDX203. Each dot represents data from one experiment and mean values of data from replicate wells from each experiment are shown. (G–J) Luciferase-labeled PDX203 5D4 tumor cells (0.2×106 /mouse) were injected via tail vein into NSG mice on day −4 and CD79b CAR T cells or untransduced T cells (5×106 /mouse) derived from two different donors were injected via tail vein on day 0. Tumor burden was monitored by bioluminescence imaging (G and I). Average radiance (photons/sec/cm2/steradian) of groups of mice on days 35 and 29 are shown (H and J). (n=5 mice per group, mean±SD is shown). ****p<0.0001; ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05. CAR, chimeric antigen receptor; PDX, patient-derived xenografts.

Journal: Journal for immunotherapy of cancer

Article Title: Chimeric antigen receptor T cells to target CD79b in B-cell lymphomas.

doi: 10.1136/jitc-2023-007515

Figure Lengend Snippet: Figure 6 Efficacy of CD79b CAR T cells against patient-derived lymphoma tumor cells. CAR T cells targeting CD79b were generated from primary healthy donor T cells, labeled with CellTrace Far Red and co-cultured with Daudi Burkitt lymphoma tumor cells or lymphoma tumor cells derived from two PDXs (PDX203 and PDX300) for 96 hours. Untransduced T cells were used as controls. (A) Histograms showing expression of CD19 and CD79b in the two tumor samples. (B) Representative flow cytometric plots illustrating proliferation of CD4+ and CD8+ CARs T cells in response to tumor cells at an effector:target (E:T) ratio of 0.6:1. (C) Summary results showing proliferation from replicate wells (N=2). (D) Summary results of cytotoxicity assay from replicate wells (N=2) showing specific lysis at E:T ratio of 0.6:1. (E and F) Pooled data from 17 experiments testing CD79b CAR T cells generated from five donors is shown for per cent change in absolute number of live tumor cells on day 4 in an in vitro cytotoxicity assay compared with day 0 (E) and fold change in interferon-γ production compared with untransduced T cells after 24 hours of co-culture (F). Target tumor cells in these experiments included Daudi, Jeko-1, SUDHL6, or PDX203. Each dot represents data from one experiment and mean values of data from replicate wells from each experiment are shown. (G–J) Luciferase-labeled PDX203 5D4 tumor cells (0.2×106 /mouse) were injected via tail vein into NSG mice on day −4 and CD79b CAR T cells or untransduced T cells (5×106 /mouse) derived from two different donors were injected via tail vein on day 0. Tumor burden was monitored by bioluminescence imaging (G and I). Average radiance (photons/sec/cm2/steradian) of groups of mice on days 35 and 29 are shown (H and J). (n=5 mice per group, mean±SD is shown). ****p<0.0001; ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05. CAR, chimeric antigen receptor; PDX, patient-derived xenografts.

Techniques: Derivative Assay, Generated, Labeling, Cell Culture, Expressing, Cytotoxicity Assay, Lysis, In Vitro, Co-Culture Assay, Luciferase, Injection, Imaging

Figure 7 CD79b CARs specially recognized human CD79b protein. (A) CAR T cells targeting CD79b were generated from primary healthy donor T cells, stained with fluorochrome labeled recombinant human CD19-Fc, CD79a-Fc, or CD79b-Fc proteins, and assessed by flow cytometry. Transduction efficiency determined by eGFP expression and binding of recombinant proteins is shown. (B–D) CAR constructs targeting CD19 or CD79b were transduced lentivirally into Jurkat-Lucia NFAT reporter cell line and cultured alone or with the indicated lymphoma cell lines at an effector:target (E:T) ratio of 1:1 or with anti-CD3 and anti-CD28 monoclonal antibodies. NFAT activation was determined after 24 hours by measuring luciferase activity. Percentage of Jurkat-Lucia NFAT reporter cells expressing CD19 or CD79b CAR was determined by assessing eGFP transduction marker by flow cytometry (B). Luciferase activity shown in Jurkat-Lucia NFAT reporter cells transduced with 79b CAR construct and cultured alone or with anti-CD3/CD28 antibodies or Daudi Burkitt lymphoma cells (C). Luciferase activity shown in Jurkat-Lucia NFAT reporter cells transduced with the indicated CD19 or CD79b CAR constructs and cultured alone or with isogenic SUDHL6 lymphoma cell lines (parent, CD19KO, CD79bKO, or CD19KOCD79bKO). (E) Cytotoxic activity of CD19 or CD79b CAR T cells generated from healthy donor T cells against isogenic cell lines of SUDHL6 (parent, CD19KO, CD79bKO, or CD19KOCD79bKO). (F) Cytotoxic activity of CD79b CAR T cells against wild type Jeko-1 and PDX203 5D4 (CD19+CD79b+), Jeko-1 CD19KO, Jeko-1 CD79bKO, or NALM6 acute lymphoblastic leukemia (CD19+CD79b–) cell lines at an E:T ratio of 1:1. The absolute number of live tumor cells at the end of culture period from this experiment are shown in online supplemental figure 10. (G and H) Luciferase- labeled Jeko-1 CD19KO or Jeko-1 CD79bKO tumor cells (0.2×106 /mouse) were injected via tail vein into NSG mice on day −4 and CD79b CAR T cells or untransduced T cells (5×106 /mouse) were injected via tail vein on day 0. Tumor burden was monitored by bioluminescence imaging at the indicated time points (G) and average radiance (photons/sec/cm2/steradian) of groups of mice on day 39 is shown (H). Data represent the mean±SD of replicate wells (N=2) where applicable and is representative of at least three separate experiments from two different donors (C–F). ****p<0.0001; ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05. CAR, chimeric antigen receptor; eGFR, enhanced green fluorescent protein; HD, healthy donor; KO, knock-out; RLU, relative luminescence units.

Journal: Journal for immunotherapy of cancer

Article Title: Chimeric antigen receptor T cells to target CD79b in B-cell lymphomas.

doi: 10.1136/jitc-2023-007515

Figure Lengend Snippet: Figure 7 CD79b CARs specially recognized human CD79b protein. (A) CAR T cells targeting CD79b were generated from primary healthy donor T cells, stained with fluorochrome labeled recombinant human CD19-Fc, CD79a-Fc, or CD79b-Fc proteins, and assessed by flow cytometry. Transduction efficiency determined by eGFP expression and binding of recombinant proteins is shown. (B–D) CAR constructs targeting CD19 or CD79b were transduced lentivirally into Jurkat-Lucia NFAT reporter cell line and cultured alone or with the indicated lymphoma cell lines at an effector:target (E:T) ratio of 1:1 or with anti-CD3 and anti-CD28 monoclonal antibodies. NFAT activation was determined after 24 hours by measuring luciferase activity. Percentage of Jurkat-Lucia NFAT reporter cells expressing CD19 or CD79b CAR was determined by assessing eGFP transduction marker by flow cytometry (B). Luciferase activity shown in Jurkat-Lucia NFAT reporter cells transduced with 79b CAR construct and cultured alone or with anti-CD3/CD28 antibodies or Daudi Burkitt lymphoma cells (C). Luciferase activity shown in Jurkat-Lucia NFAT reporter cells transduced with the indicated CD19 or CD79b CAR constructs and cultured alone or with isogenic SUDHL6 lymphoma cell lines (parent, CD19KO, CD79bKO, or CD19KOCD79bKO). (E) Cytotoxic activity of CD19 or CD79b CAR T cells generated from healthy donor T cells against isogenic cell lines of SUDHL6 (parent, CD19KO, CD79bKO, or CD19KOCD79bKO). (F) Cytotoxic activity of CD79b CAR T cells against wild type Jeko-1 and PDX203 5D4 (CD19+CD79b+), Jeko-1 CD19KO, Jeko-1 CD79bKO, or NALM6 acute lymphoblastic leukemia (CD19+CD79b–) cell lines at an E:T ratio of 1:1. The absolute number of live tumor cells at the end of culture period from this experiment are shown in online supplemental figure 10. (G and H) Luciferase- labeled Jeko-1 CD19KO or Jeko-1 CD79bKO tumor cells (0.2×106 /mouse) were injected via tail vein into NSG mice on day −4 and CD79b CAR T cells or untransduced T cells (5×106 /mouse) were injected via tail vein on day 0. Tumor burden was monitored by bioluminescence imaging at the indicated time points (G) and average radiance (photons/sec/cm2/steradian) of groups of mice on day 39 is shown (H). Data represent the mean±SD of replicate wells (N=2) where applicable and is representative of at least three separate experiments from two different donors (C–F). ****p<0.0001; ***p<0.001; **p<0.01; *p<0.05; (ns) p>0.05. CAR, chimeric antigen receptor; eGFR, enhanced green fluorescent protein; HD, healthy donor; KO, knock-out; RLU, relative luminescence units.

Techniques: Generated, Staining, Labeling, Recombinant, Flow Cytometry, Transduction, Expressing, Binding Assay, Construct, Cell Culture, Bioprocessing, Activation Assay, Luciferase, Activity Assay, Marker, Injection, Imaging, Knock-Out

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