Journal: Antibodies

Article Title: Adoptive Cell Therapy in Mice Sensitized to a Grass Pollen Allergen

doi: 10.3390/antib13020048

Figure Lengend Snippet: Experimental Groups (ATG, anti-thymocyte globulin; BMC, bone marrow cells; immune, immunization).

Article Snippet: Additionally, mice received costimulation blockade consisting of anti-CD40L (MR1; 1 mg, day 0; BioXcell, Lebanon, NH, USA) and CTLA4Ig (0.5 mg, day 2; abatacept, Bristol-Myers Squibb Pharmaceuticals, Princeton, NJ, USA).

Techniques: Irradiation

Journal: Antibodies

Article Title: Adoptive Cell Therapy in Mice Sensitized to a Grass Pollen Allergen

doi: 10.3390/antib13020048

Figure Lengend Snippet: Experimental Timeline BALB/c mice were sensitized to Phl p 5 and Bet v 1 in weeks −15 and −12 before and 4 and 7 weeks after cell therapy. On day 0, 25 × 10 6 Phl p 5 + BM cells were injected i.v. One experimental group received a second dose of 25 × 10 6 BM cells on day 5. Mice were pre-treated with a single dose of ATG (0.45 mg, day −2) and 2Gy irradiation on day −1. Additionally, they received costimulation blockade (1 mg MR1, 0.5 mg CTLA4Ig) and a short course of Rapamycin (0.1 mg, day −1, d0, d2). 17 weeks after treatment, mice received a Phl p 5 + skin allograft and 22 weeks later mice were sacrificed for in vitro analysis of Phl p 5-specific T cell proliferation. Remaining skin grafts were histologically analysed. (ATG, anti-thymocyte globulin; BMC, bone marrow cells; sc, subcutaneous).

Article Snippet: Additionally, mice received costimulation blockade consisting of anti-CD40L (MR1; 1 mg, day 0; BioXcell, Lebanon, NH, USA) and CTLA4Ig (0.5 mg, day 2; abatacept, Bristol-Myers Squibb Pharmaceuticals, Princeton, NJ, USA).

Techniques: Injection, Irradiation, In Vitro

Journal: Nature Communications

Article Title: Implantable niche with local immunosuppression for islet allotransplantation achieves type 1 diabetes reversal in rats

doi: 10.1038/s41467-022-35629-z

Figure Lengend Snippet: a Optical image of NICHE and annotated rendering of NICHE and scanning electron microscopy (SEM) images of the two-layer mesh and nanoporous membrane (representative scan images of 5 independent samples). The longitudinal section of NICHE is representative for clarity. Representative Masson’s trichrome staining of fibrotic capsule around b resin and c PA devices implanted in rats for 6 weeks. d Quantification of fibrotic capsule thickness around medical grade titanium (Ti; n = 5), resin ( n = 5), and PA ( n = 4) devices implanted subQ for 6 weeks in rats, mean ± SD, one-way analysis of variance (ANOVA) *** p < 0.001. e Implant r e activity scores of Ti ( n = 5), resi n ( n = 5), and PA ( n = 4) devices implanted subQ for 6 weeks in rats, mean ± SD, one-way ANOVA, n.s. p = 0.1117. Optical images showing cross sections of f resin and g PA NICHE implanted subQ for 6 weeks. Dashed lines indicate cell reservoir. h Resin and PA roughness quantified using atomic force microscopy ( n = 9 devices per material), mean ± SD, unpaired two-tailed student’s t-test ** p < 0.01. i Contact angle analysis of resin and PA devices ( n = 3 devices per material), mean ± SD, unpaired two-tailed student’s t-test *** p < 0.001. j Resin and PA NICHE cell reservoir tissue sections stained with blood vessel marker B. simplicifolia lectin; vessels in red pointed by black arrow heads. k Vascular density quantification of resin ( n = 1) and PA ( n = 4) cell reservoir tissue from independent NICHE, mean ± SD. l Daily percent in vitro release of CTLA4Ig and IgG from NICHE drug reservoir ( n = 4 devices per drug), mean ± SD. Static glucose stimulated insulin release (GSIR) of rat islets after 5 days of incubation with m CTLA4Ig, n ALS, and o CTLA4Ig + ALS ( n = 4 per condition), mean ± SD, o n e-way ANOVA, * p < 0.05, ** p < 0.001, n.s. p > 0.05 of each condition versus vehicle (0 mg/mL). Source data are provided as a Source Data file.

Article Snippet: The immunosuppressants used for rat studies were CTLA4Ig (Abatacept, Bristol-Myers Squibb) and rabbit anti-lymphocyte serum (ALS; Accurate chemical); and for NHP studies anti-thymocyte globulin (ATG; Sanofi), CTLA4Ig (Belatacept, Bristol-Myers Squibb), and anti-CD154 [5C8H1] (NIH Nonhuman Primate Reagent Resource, RRID: AB_2716324).

Techniques: Electron Microscopy, Membrane, Staining, Activity Assay, Microscopy, Two Tailed Test, Marker, In Vitro, Incubation

Journal: Nature Communications

Article Title: Implantable niche with local immunosuppression for islet allotransplantation achieves type 1 diabetes reversal in rats

doi: 10.1038/s41467-022-35629-z

Figure Lengend Snippet: a In vitro tube formation activity of human umbilical vein endothelial cells (HUVEC) in the presence of different drug concentrations (mg/mL). Supplemented media was used as positive inducer (+), 30 µM suramin as positive inhibitor (-), and non-supplemented media as control (Veh). b Total length quantification and c number of segments of tube formation ( n = 4 biological replicates per condition), mean ± SD, one-way ANOVA with Tukey’s multiple comparisons test (*** p < 0.001). d qPCR analysis of vascular related genes in HUVEC after treatment with different concentrations of ATG and CTLA4Ig (mg/mL). Gapdh gene was used as internal control and fold-change in gene expression is relative to control untreated cells. ( n = 4 biological replicates), mean ± SD, one-way ANOVA with Tukey’s multiple comparisons test (*** p < 0.001; n.s. p > 0.05). e Quantification of VEGF levels in cell reservoir and surrounding microenvironment for control and IS release samples ( n = 4 biological replicates), mea n ± SD, unpaired two-tailed student’s t -test n.s. p > 0.05. f Fluorescence intensity measurements of IHC analysis in g ( n = 4 biological replicates), mean ± SD, unpaired two-tailed student’s t -test n.s. p > 0.05. g Representative sections of NICHE control and IS release slides stained with functional blood vessel markers CD31 (red), VE-cadherin (green), and eNOS (gold). Source data are provided as a Source Data file.

Article Snippet: The immunosuppressants used for rat studies were CTLA4Ig (Abatacept, Bristol-Myers Squibb) and rabbit anti-lymphocyte serum (ALS; Accurate chemical); and for NHP studies anti-thymocyte globulin (ATG; Sanofi), CTLA4Ig (Belatacept, Bristol-Myers Squibb), and anti-CD154 [5C8H1] (NIH Nonhuman Primate Reagent Resource, RRID: AB_2716324).

Techniques: In Vitro, Activity Assay, Control, Gene Expression, Two Tailed Test, Fluorescence, Staining, Functional Assay

Journal: Nature Communications

Article Title: Implantable niche with local immunosuppression for islet allotransplantation achieves type 1 diabetes reversal in rats

doi: 10.1038/s41467-022-35629-z

Figure Lengend Snippet: Plasma levels of a ALS and b CTLA4Ig of NICHE ( n = 12 to day 63, n = 8 to day 92, n = 5 to day 112, n = 3 to day 192) and IP ( n = 8 to day 63, n = 5 to day 112) rats, mean ± SD, unpaired two-tailed student’s t -test between NICHE and IP overall average concentrations, (*** p < 0.001). Black triangles indicate drug reservoir reloading. The red triangle indicates systemic ALS bolus. LOQ limit of quantitation. Quantification of immunosuppressants in c peripheral organs and d transplant microenvironment at endpoint of NICHE and IP rats ( n = 8), mea n ± SD, unpaired two-tailed student’s t -test between NICHE and IP rats for each organ (* p < 0.05; ** p < 0.01; *** p < 0.001; n.s. p > 0.05; n.q. = not quantifiable). FC = fibrotic capsule. e Quantification of lymphocytes in blood expressed as fold change to day 0 of NICHE ( n = 8 to day 56, n = 5 to day 115), IP ( n = 5), and rats receiving vehicle ( n = 4). f Quantification of Tregs in blood on day 84 of NICHE, IP, and healthy rats ( n = 5), mean ± SD, one-way ANOVA with Tukey’s multiple comparisons test (*** p < 0.001; n.s. p = 0.1643). Source data are provided as a Source Data file.

Article Snippet: The immunosuppressants used for rat studies were CTLA4Ig (Abatacept, Bristol-Myers Squibb) and rabbit anti-lymphocyte serum (ALS; Accurate chemical); and for NHP studies anti-thymocyte globulin (ATG; Sanofi), CTLA4Ig (Belatacept, Bristol-Myers Squibb), and anti-CD154 [5C8H1] (NIH Nonhuman Primate Reagent Resource, RRID: AB_2716324).

Techniques: Clinical Proteomics, Two Tailed Test, Quantitation Assay

Journal: JCI Insight

Article Title: T cell–derived tumor necrosis factor induces cytotoxicity by activating RIPK1-dependent target cell death

doi: 10.1172/jci.insight.148643

Figure Lengend Snippet: ( A – D ) Kaplan-Meier survival curves for WT or genetically distinct B6 heart grafts subjected to 8 hours (8h) of CIS and transplanted into CTLA4Ig-treated BALB/c recipients. ( A ) WT B6 hearts into recipients treated with anti-TNF mAb ( n = 5) or isotype IgG ( n = 3) control on the day of surgery. ( B ) WT B6 hearts into recipients treated with necrostatin-1 ( n = 8) or vehicle control ( n = 4) on the day of surgery. ( C ) Ripk1 D138N ( n = 6), Mlkl –/– ( n = 6), Ripk3 –/– ( n = 5), or WT ( n = 8) B6 allografts. * P < 0.05 versus WT control. ( D ) WT (same data as shown in C ) or Sharpin cpdm B6 allografts transplanted into WT BALB/c recipients ± anti-TNF blocking mAb (αTNF, day 0, then every 5 days for 2 weeks, n = 5) or equivalent dosing of IgG isotype control (isotype n = 4, P < 0.05 versus IgG control). ( E ) Representative TUNEL staining (arrowheads) and quantification of day-7 posttransplant WT and RIPK1 D138N B6 hearts in WT BALB/c recipients ( n = 4–5). Each dot represents a biological replicate. P < 0.05 versus WT control, t test. ( F ) Representative TUNEL staining and quantification of WT B6 allografts transplanted as described above into BALB/c hosts treated with anti-TNF mAb or isotype control ( n = 5–6, t test). ( G ) Representative TUNEL staining and quantification of Sharpin cpdm B6 allografts transplanted as described above into BALB/c hosts treated with anti-TNF mAb or isotype control ( n = 4–5). * P < 0.05. Scale bars: 100 μm. Survival statistics for A – D calculated by Mantel Cox log rank test. Quantitative statistics for E – G calculated by t test.

Article Snippet: Recipient mice received 250 μg CTLA4Ig (Orencia, Bristol Myers Squibb) i.p. on the day of surgery when indicated.

Techniques: Control, Blocking Assay, TUNEL Assay, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Alloantibodies Prevent the Induction of Transplantation Tolerance by Enhancing Alloreactive T cell Priming

doi: 10.4049/jimmunol.1001172

Figure Lengend Snippet: CTLA4Ig overrides the effects of Anti-Kd mAbs and facilitates graft acceptance in anti-CD154/DST-treated recipients. Acceptance of BALB/c skin grafts by B6 recipients treated with anti-CD154/DST + CTLA4Ig (N =4) (closed square) anti-CD154/DST + CTLA4Ig with anti-Kd mAbs (N = 7) (closed downward triangle) but rejection by B6 recipients treated with anti-CD154/DST and anti-Kd mAbs (N = 15) (open upward triangle). B6 recipients treated with anti-CD154/DST (N = 11) (closed circle) also accepted BALB/c skin grafts.

Article Snippet: Human CTLA4Ig (huCTLA4Ig; 0.5mg /mouse) (Bristol-Myers Squibb; New York City, NY) was administered daily, by intra-peritoneal (i.p.) injection, for 7 days post-skin transplantation.

Techniques: