Journal: bioRxiv

Article Title: Inhibition of De Novo Autoantibody Production Attenuates Chronic Antibody-Mediated Rejection of Kidney Allografts Despite Maintenance of High Donor-Specific Antibody Titers

doi: 10.1101/2025.06.11.659154

Figure Lengend Snippet: A. Protocol of anti-CD20mAb treatment. Groups of B6.CCR5 -/- /cG -/- mice were transplanted with A/J kidney allografts and treated with 250 ug anti-mouse CD20 mAb i.p on days 5, 8 and 12 post-transplant. B. Graft survival and C. DSA titer were determined at the indicated time points post-transplant. D. the presence of CD19 + B220 + B cells in the peripheral blood, spleen and kidney graft of anti-CD20 mAb or non-treated recipients was assessed with significant decreases in the peripheral blood and kidney allograft but not in the spleen. E. Blood urea nitrogen (BUN) in serum samples from anti-CD20 mAb treated and non-treated B6.CCR5 -/- /cG -/- kidney allograft recipients as well as in wild type C57BL/6 kidney iso-and allo-graft recipients were assessed on day 60 after transplant.

Article Snippet: As indicated, B6.CCR5/cG -/- kidney allograft recipients were treated with 250 μg anti-mouse CD20 monoclonal antibody (Clone: AISB12, Catalog# BE0302, BioXCell, Lebanon, NH) i.p. on days 5, 8 and 12 after transplant or with 100 μg anti-mouse CD19 (Clone: 1D3, Catalog# BE0150, BioXCell) plus 100 μg B220 monoclonal antibody (Clone: RA3.3A1/6.1, Catalog# BE0067, BioXCell) i.p. either on days 14, 19, 24 and 29 (early) or on days 34, 39, 44 and 49 (late) post-transplant.

Techniques:

Journal: bioRxiv

Article Title: Inhibition of De Novo Autoantibody Production Attenuates Chronic Antibody-Mediated Rejection of Kidney Allografts Despite Maintenance of High Donor-Specific Antibody Titers

doi: 10.1101/2025.06.11.659154

Figure Lengend Snippet: A. Protocol of anti-CD19 plus anti-B220 mAb treatment. A/J kidneys were transplanted to B6.CCR5/cG -/- mice that were treated with 200 ug (100 ug each) of anti-CD19 plus B220 mAb i.p. on either days 14, 19, 24 and 29 or days 34, 39, 44 and 49. B. B cell population changes by the anti-CD19mAb plus B220 mAb treatment in peripheral blood. B cell was completely depleted after treatment and 20% was back on day 60. C. Groups of 5 B6.CCR5/cG -/- mice were transplanted with A/J kidney allografts and treated with 200 ug i.p. anti-CD19 plus B220 mAb on either days 14, 19, 24 and 29 or on days 34, 39, 44, and 49 and graft survival was assessed. D. Serum was obtained from the recipients in on days 16, 35 and 60 and the DSA titers determined.

Article Snippet: As indicated, B6.CCR5/cG -/- kidney allograft recipients were treated with 250 μg anti-mouse CD20 monoclonal antibody (Clone: AISB12, Catalog# BE0302, BioXCell, Lebanon, NH) i.p. on days 5, 8 and 12 after transplant or with 100 μg anti-mouse CD19 (Clone: 1D3, Catalog# BE0150, BioXCell) plus 100 μg B220 monoclonal antibody (Clone: RA3.3A1/6.1, Catalog# BE0067, BioXCell) i.p. either on days 14, 19, 24 and 29 (early) or on days 34, 39, 44 and 49 (late) post-transplant.

Techniques:

Journal: bioRxiv

Article Title: Inhibition of De Novo Autoantibody Production Attenuates Chronic Antibody-Mediated Rejection of Kidney Allografts Despite Maintenance of High Donor-Specific Antibody Titers

doi: 10.1101/2025.06.11.659154

Figure Lengend Snippet: A/J kidneys were transplanted to B6.CCR5/cG -/- mice that were treated with 200 ug (100 ug each) of anti-CD19 plus B220 mAb i.p. on either days 14, 19, 24 and 29 or days 34, 39, 44 and 49. Allografts were harvested at day 60 and sections prepared for histological evaluation: A. C4d; and, B. α smooth muscle actin. C. Graft homogenates were prepared on day 60 post-transplanted and tested for quantitation of hydroxyproline as an indication of collagen content per mg of wet graft tissue. *p < 0.05. D. Serum was obtained from recipients at day 60 post-transplant and blood urea nitrogen quantitated. *p < 0.05. E. Serum obtained on day 60 post-transplant from each recipient was assessed for the presence of auto-antibodies to extra-cellular matrix (ECM) proteins using an Auto-Antibody array.

Article Snippet: As indicated, B6.CCR5/cG -/- kidney allograft recipients were treated with 250 μg anti-mouse CD20 monoclonal antibody (Clone: AISB12, Catalog# BE0302, BioXCell, Lebanon, NH) i.p. on days 5, 8 and 12 after transplant or with 100 μg anti-mouse CD19 (Clone: 1D3, Catalog# BE0150, BioXCell) plus 100 μg B220 monoclonal antibody (Clone: RA3.3A1/6.1, Catalog# BE0067, BioXCell) i.p. either on days 14, 19, 24 and 29 (early) or on days 34, 39, 44 and 49 (late) post-transplant.

Techniques: Quantitation Assay, Ab Array

Journal: bioRxiv

Article Title: Inhibition of De Novo Autoantibody Production Attenuates Chronic Antibody-Mediated Rejection of Kidney Allografts Despite Maintenance of High Donor-Specific Antibody Titers

doi: 10.1101/2025.06.11.659154

Figure Lengend Snippet: A. A/J kidneys were transplanted to groups of 3 B6.CCR5/cG -/- recipients that were untreated, treated with 200 ug anti-mouse CD20 on days 5, 8 and 12, or treated with 200 ug (100 ug each) of anti-CD19 plus B220 mAb i.p. on either days 14, 19, 24 and 29 (Early) or days 34, 39, 44 and 49 (Late). Allografts were harvested on day 60 and graft RNA was isolated and analyzed by the NanoString nCounter platform using the Mouse Fibrosis panel, and heatmaps were generated from the top differentially expressed genes for comparison between the allografts from the 4 groups of B6.CCR5/cG -/- recipients. B. Volcano plot and heatmap show the differently expressed genes between kidney allografts harvested from untreated B6.CCR5/cG -/- recipients vs. B6.CCR5/cG -/- recipients treated with the early anti-CD19 plus B220 mAb strategy. The horizontal dotted line on the volcano plot is p value = 0.05. The two vertical dotted lines are log 2 fold-change-1 and 1. Red dots indicate DEGs with increased expression in allografts from untreated B6.CCR5/cG -/- recipients and blue dots indicate DEGs with decreased expression in allografts treated with the Early B cell depletion strategy. C. Volcano plot and heatmap show the differently expressed genes between kidney allografts harvested from untreated B6.CCR5/cG -/- recipients vs. B6.CCR5/cG -/- recipients treated with the Late anti-CD19 plus B220 mAb strategy. The horizontal dotted line on the volcano plot is p value = 0.05. The two vertical dotted lines are log 2 fold-change-1 and 1. Red dots indicate DEGs with increased expression in allografts from untreated B6.CCR5/cG -/- recipients and blue dots indicate DEGs with decreased expression in allografts treated with the Late B cell depletion strategy. D. Volcano plot show the differently expressed genes between kidney allografts harvested from B6.CCR5/cG -/- recipients treated with the Early B cell strategy vs. with the Late B cell depletion strategy. The horizontal dotted line on the volcano plot is p value = 0.05. The two vertical dotted lines are log 2 fold-change-1 and 1. Red dots indicate DEGs with increased expression in allografts from B6.CCR5/cG -/- recipients treated with the Early B cell depletion strategy and blue dots indicate DEGs with decreased expression in allografts treated with the Late B cell depletion strategy.

Article Snippet: As indicated, B6.CCR5/cG -/- kidney allograft recipients were treated with 250 μg anti-mouse CD20 monoclonal antibody (Clone: AISB12, Catalog# BE0302, BioXCell, Lebanon, NH) i.p. on days 5, 8 and 12 after transplant or with 100 μg anti-mouse CD19 (Clone: 1D3, Catalog# BE0150, BioXCell) plus 100 μg B220 monoclonal antibody (Clone: RA3.3A1/6.1, Catalog# BE0067, BioXCell) i.p. either on days 14, 19, 24 and 29 (early) or on days 34, 39, 44 and 49 (late) post-transplant.

Techniques: Isolation, Generated, Comparison, Expressing