Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: (A) Galectin-family protein groups, including an LGALS13-annotated / CLC-Galectin-10-like porcine orthologous protein group. (B) Immune lineage-associated proteins (T-cell, B-cell, myeloid, cytotoxic, and pan-leukocyte markers). (C) Selected signaling protein groups (NF-κB and JAK/STAT axes). DIA proteomics was performed on effluent cell pellets from one representative biological perfusion in technical triplicate and should be interpreted as exploratory cell-associated protein abundance. Values are log2 fold-changes relative to the Pre-1 baseline; heatmap scale capped at ±3 for display. UD = undetected.

Article Snippet: Human Galectin-10 induced marked apoptosis of CD3/CD28-stimulated Jurkat cells, prioritizing this axis for future mechanistic testing without proving causality in the perfused spleen.

Techniques: Quantitative Proteomics

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: Jurkat T cells were stimulated with anti-CD3/CD28 (5 µg/mL) for 1 h before addition of recombinant human Galectin-10 or Galectin-13 (20 µg/mL) for 24 h. Cells were stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. Stacked bars decompose the total Annexin V+ population into early-apoptotic (Annexin V+ / PI−, gold) and late-apoptotic / dying (Annexin V+ / PI+, red) fractions. Total Annexin V+ values are annotated above each bar; error bars represent SEM on the total. Total Annexin V+ values were compared across conditions by one-way ANOVA with Dunnett’s multiple-comparison test against the stimulated control. Individual replicate values are shown as overlaid points (n = 3 per condition). Human Galectin-10 produced ∼84% total Annexin V+ positivity, the great majority of which had progressed to the late-apoptotic / dying state by 24 h compared with stimulated alone (p<0.001). Galectin-13 produced a more modest increase. This assay was selected because the porcine LGALS13-annotated proteomic signal is interpreted as a CLC/Galectin-10-like orthologous axis; it supports prioritization of that axis but does not prove porcine protein identity or causality in the perfused spleen.

Article Snippet: Human Galectin-10 induced marked apoptosis of CD3/CD28-stimulated Jurkat cells, prioritizing this axis for future mechanistic testing without proving causality in the perfused spleen.

Techniques: Recombinant, Staining, Flow Cytometry, Comparison, Control, Produced

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: (A) Galectin-family protein groups, including an LGALS13-annotated / CLC-Galectin-10-like porcine orthologous protein group. (B) Immune lineage-associated proteins (T-cell, B-cell, myeloid, cytotoxic, and pan-leukocyte markers). (C) Selected signaling protein groups (NF-κB and JAK/STAT axes). DIA proteomics was performed on effluent cell pellets from one representative biological perfusion in technical triplicate and should be interpreted as exploratory cell-associated protein abundance. Values are log2 fold-changes relative to the Pre-1 baseline; heatmap scale capped at ±3 for display. UD = undetected.

Article Snippet: – Recombinant human Galectin-10 produced a marked increase in Annexin V positivity and Annexin V/PI double-positivity.

Techniques: Quantitative Proteomics

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: Jurkat T cells were stimulated with anti-CD3/CD28 (5 µg/mL) for 1 h before addition of recombinant human Galectin-10 or Galectin-13 (20 µg/mL) for 24 h. Cells were stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. Stacked bars decompose the total Annexin V+ population into early-apoptotic (Annexin V+ / PI−, gold) and late-apoptotic / dying (Annexin V+ / PI+, red) fractions. Total Annexin V+ values are annotated above each bar; error bars represent SEM on the total. Total Annexin V+ values were compared across conditions by one-way ANOVA with Dunnett’s multiple-comparison test against the stimulated control. Individual replicate values are shown as overlaid points (n = 3 per condition). Human Galectin-10 produced ∼84% total Annexin V+ positivity, the great majority of which had progressed to the late-apoptotic / dying state by 24 h compared with stimulated alone (p<0.001). Galectin-13 produced a more modest increase. This assay was selected because the porcine LGALS13-annotated proteomic signal is interpreted as a CLC/Galectin-10-like orthologous axis; it supports prioritization of that axis but does not prove porcine protein identity or causality in the perfused spleen.

Article Snippet: – Recombinant human Galectin-10 produced a marked increase in Annexin V positivity and Annexin V/PI double-positivity.

Techniques: Recombinant, Staining, Flow Cytometry, Comparison, Control, Produced

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: (A) Galectin-family protein groups, including an LGALS13-annotated / CLC-Galectin-10-like porcine orthologous protein group. (B) Immune lineage-associated proteins (T-cell, B-cell, myeloid, cytotoxic, and pan-leukocyte markers). (C) Selected signaling protein groups (NF-κB and JAK/STAT axes). DIA proteomics was performed on effluent cell pellets from one representative biological perfusion in technical triplicate and should be interpreted as exploratory cell-associated protein abundance. Values are log2 fold-changes relative to the Pre-1 baseline; heatmap scale capped at ±3 for display. UD = undetected.

Article Snippet: The human Galectin-10 Jurkat assay was performed because the porcine DIA hit was annotated as LGALS13 but interpreted as an orthologous CLC/Galectin-10-like signal with a plausible human immune counterpart.

Techniques: Quantitative Proteomics

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: Jurkat T cells were stimulated with anti-CD3/CD28 (5 µg/mL) for 1 h before addition of recombinant human Galectin-10 or Galectin-13 (20 µg/mL) for 24 h. Cells were stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. Stacked bars decompose the total Annexin V+ population into early-apoptotic (Annexin V+ / PI−, gold) and late-apoptotic / dying (Annexin V+ / PI+, red) fractions. Total Annexin V+ values are annotated above each bar; error bars represent SEM on the total. Total Annexin V+ values were compared across conditions by one-way ANOVA with Dunnett’s multiple-comparison test against the stimulated control. Individual replicate values are shown as overlaid points (n = 3 per condition). Human Galectin-10 produced ∼84% total Annexin V+ positivity, the great majority of which had progressed to the late-apoptotic / dying state by 24 h compared with stimulated alone (p<0.001). Galectin-13 produced a more modest increase. This assay was selected because the porcine LGALS13-annotated proteomic signal is interpreted as a CLC/Galectin-10-like orthologous axis; it supports prioritization of that axis but does not prove porcine protein identity or causality in the perfused spleen.

Article Snippet: The human Galectin-10 Jurkat assay was performed because the porcine DIA hit was annotated as LGALS13 but interpreted as an orthologous CLC/Galectin-10-like signal with a plausible human immune counterpart.

Techniques: Recombinant, Staining, Flow Cytometry, Comparison, Control, Produced

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: (A) Galectin-family protein groups, including an LGALS13-annotated / CLC-Galectin-10-like porcine orthologous protein group. (B) Immune lineage-associated proteins (T-cell, B-cell, myeloid, cytotoxic, and pan-leukocyte markers). (C) Selected signaling protein groups (NF-κB and JAK/STAT axes). DIA proteomics was performed on effluent cell pellets from one representative biological perfusion in technical triplicate and should be interpreted as exploratory cell-associated protein abundance. Values are log2 fold-changes relative to the Pre-1 baseline; heatmap scale capped at ±3 for display. UD = undetected.

Article Snippet: Annexin V+ / PI+ double-positive cells increased from 3.7 ± 0.3% in resting cells and 12.1 ± 0.5% in stimulated controls to 80.7 ± 7.0% with human Galectin-10 ( ).

Techniques: Quantitative Proteomics

Journal: bioRxiv

Article Title: Acellular normothermic spleen perfusion resolves transcriptional and non-transcriptional mechanisms of steroid immunosuppression

doi: 10.64898/2026.05.16.725632

Figure Lengend Snippet: Jurkat T cells were stimulated with anti-CD3/CD28 (5 µg/mL) for 1 h before addition of recombinant human Galectin-10 or Galectin-13 (20 µg/mL) for 24 h. Cells were stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. Stacked bars decompose the total Annexin V+ population into early-apoptotic (Annexin V+ / PI−, gold) and late-apoptotic / dying (Annexin V+ / PI+, red) fractions. Total Annexin V+ values are annotated above each bar; error bars represent SEM on the total. Total Annexin V+ values were compared across conditions by one-way ANOVA with Dunnett’s multiple-comparison test against the stimulated control. Individual replicate values are shown as overlaid points (n = 3 per condition). Human Galectin-10 produced ∼84% total Annexin V+ positivity, the great majority of which had progressed to the late-apoptotic / dying state by 24 h compared with stimulated alone (p<0.001). Galectin-13 produced a more modest increase. This assay was selected because the porcine LGALS13-annotated proteomic signal is interpreted as a CLC/Galectin-10-like orthologous axis; it supports prioritization of that axis but does not prove porcine protein identity or causality in the perfused spleen.

Article Snippet: Annexin V+ / PI+ double-positive cells increased from 3.7 ± 0.3% in resting cells and 12.1 ± 0.5% in stimulated controls to 80.7 ± 7.0% with human Galectin-10 ( ).

Techniques: Recombinant, Staining, Flow Cytometry, Comparison, Control, Produced