Journal: Journal of Human Immunity

Article Title: Increased prevalence of autoantibodies neutralizing IFNλ2/3 in young individuals with cystic fibrosis

doi: 10.70962/jhi.20250268

Figure Lengend Snippet: At least 10% of young individuals with CF harbor neutralizing anti-IFNλ2/3 IgG autoAbs. (A) IFNλ1/2/3 neutralization by plasmas from individuals with AD ( n = 39), various adults (CT; n = 68), or pwCF (CF; n = 84 samples from n = 51 individuals). Dotted lines indicate thresholds set using standard outlier calculations. Neutralizing plasmas are orange; numbers represent unique individuals; positive individuals are labeled. (B) Anti-IFN-III and anti-IFN-I IgGs in CF plasmas positive (neut; n = 9) or negative (non-neut; n = 6) for IFNλ2/3 neutralization. (C) Spearman’s correlation between IFNλ2/3 neutralization (A) and IgG (B). (D) Anti-IFNλ2/3 IgG subclasses in CF plasmas ( n = 5). (E) Area under the curve from PIV5-GFP replication kinetics in A549 cells pretreated with IFNλ2 (or not) in the presence of pwCF plasmas positive (pos) or negative (neg) for IFNλ2/3 neutralization. Values are triplicate means from two experiments. Error bars indicate standard deviations. For the IFNλ2+pos and IFNλ2+neg groups, data are from two patients (squares/triangles) from two experiments. (F) Fluorescence images of infected cells from E. (G) Random forest regression of clinical variables in pwCF positive ( n = 4) or negative ( n = 16) for anti-IFNλ2/3 autoAbs. Dots represent importance scores per iteration (maximum 5,000); error bars reflect importance spread. Abbreviations: MFI FC, median fluorescence intensity fold-change; IVIG, intravenous immunoglobulin; IBD, inflammatory bowel disease; CFRD, CF-related diabetes; ABPA, allergic bronchopulmonary aspergillosis; SA, Staphylococcus aureus ; PA, Pseudomonas aeruginosa . Statistical analyses were two-way ANOVA with the Šidák correction (B), Spearman correlation (C), and Mann–Whitney test (E) (ns, nonsignificant; *P < 0.05; ****P < 0.0001).

Article Snippet: Protein targets were: IFNλ1 (NBP2-34996; Novusbio), IFNλ2 (8417-IL; R&D Systems), IFNλ3 (5259-IL; R&D Systems), IFNα2 (NBP2-34971, Novusbio), and IFNβ (11420-1; PBL Assay Science).

Techniques: Neutralization, Labeling, Fluorescence, Infection, MANN-WHITNEY

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Muscle stem cells contribute to long‐term tissue repletion following surgical sepsis

doi: 10.1002/jcsm.13214

Figure Lengend Snippet: Satellite cell reduction reduces muscle regeneration capacity in cecal ligation and puncture/daily chronic stress (CLP/DCS) mice. ( A ) Timeline representing male Pax7‐DTA mice treated for five consecutive daily injections of tamoxifen or vehicle, 15 days before surgery. Percentage change from baseline of ( B ) lean mass. ( C ) Immunofluorescence images (top) and quantification of minimum Feret diameter (bottom) of tibialis anterior (TA) cross sections from either vehicle or tamoxifen control and CLP/DCS mice. Green = laminin, Blue = DAPI. Images are ×20 with scale bar = 200 μm. ( D ) Pax7 staining (top) and quantification of Pax7‐positive nuclei per myofibre (bottom) of tibialis anterior (TA) cross‐sections from vehicle or tamoxifen control and CLP/DCS mice. Purple = haematoxylin (nuclei), Red = Pax7. Images are ×40 with scale bar = 50 μm. All images are representative. B = Two‐way ANOVA with Bonferroni multiple comparisons (BMC) where *significant vs. control vehicle and control tamoxifen, ^significant vs. control tamoxifen, and % significant vs. control vehicle, P < 0.05. C = nonlinear fit pairwise comparisons. D = One‐way ANOVA with BMC where significance = * P < 0.05, ** P < 0.01, *** P < 0.001. See Tables S4 and S5 for all P values from all comparisons in ( B ) and ( C ), respectively. N = 3–7. Each n = one mouse.

Article Snippet: Permeabilization was performed using 0.5% Triton X‐100 (Sigma) in PBS for 5 min at room temperature (RT), then washed ×2 in immunofluorescence buffer (IFB: 3% bovine serum albumin [BSA, GoldBio], 0.2% Triton X‐100 (Sigma), 0.2% Tween‐20 [Sigma], in PBS).

Techniques: Ligation, Immunofluorescence, Control, Staining

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Muscle stem cells contribute to long‐term tissue repletion following surgical sepsis

doi: 10.1002/jcsm.13214

Figure Lengend Snippet: Characterization of muscle loss and recovery in CLP/DCS mice. ( A ) Combined cecal ligation and puncture (CLP)/daily chronic stress (DCS) Kaplan–Meier survival curve versus control mice to experiment endpoint. ( B ) Serum levels of mouse C‐reactive protein (CRP) and aspartate aminotransferase (AST) activity. ( C ) Control and CLP/DCS percentage change from baseline in lean mass. Data from mouse MRI on Day 3 or 4 are represented on lean mass graph as Day 3. ( D ) Harvested wet weights, as change in percentage of control, of tibialis anterior (TA, top) and gastrocnemius (GR, bottom) normalized to baseline body weight, respectively, in control and CLP/DCS Days 4, 8 and 28. ( E ) Immunofluorescence images (top) and quantification of minimum Feret diameter (bottom) of tibialis anterior (TA) cross‐sections from control, Days 4, 8 and 28 CLP/DCS mice. Green = laminin, Blue = DAPI. Images are representative and ×20 with scale bar = 200 μm. See Table S1 for all P value comparisons. ( F ) Immunofluorescence images (top) and quantification (bottom) of tibialis anterior (TA) myofibre typing from control, Days 4, 8 and 28 CLP/DCS mice. Red = type IIb, Green = type IIa, Unstained = type IIx, White = laminin. Images are representative and ×20 with scale bar = 100 μm. Data from muscle mass/minimum Feret diameter/myofibre typing were harvested on Day 27 or 28 and are labelled as Day 28. B = log‐rank test. C , E , G = One‐way ANOVA with Bonferroni multiple comparisons (BMC). D = Two‐way ANOVA with BMC. F = nonlinear fit pairwise comparisons. Significance: * P < 0.05, **/ & P < 0.01, ***/ # P < 0.001. ( B ) n = 12–13. ( C–D ) n = 16–45. ( E ) n = 11–14. ( F ) n = 5. ( G ) n = 10–14. Each n = one mouse.

Article Snippet: Permeabilization was performed using 0.5% Triton X‐100 (Sigma) in PBS for 5 min at room temperature (RT), then washed ×2 in immunofluorescence buffer (IFB: 3% bovine serum albumin [BSA, GoldBio], 0.2% Triton X‐100 (Sigma), 0.2% Tween‐20 [Sigma], in PBS).

Techniques: Ligation, Control, Activity Assay, Immunofluorescence

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Muscle stem cells contribute to long‐term tissue repletion following surgical sepsis

doi: 10.1002/jcsm.13214

Figure Lengend Snippet: Muscle recovery deficit in post‐sepsis mice with secondary muscle injury. ( A ) Timeline depicting experimental setup of second insult to muscle in control and cecal ligation and puncture/daily chronic stress (CLP/DCS) mice. ( B ) Immunofluorescence images (top) and minimum Feret diameter quantification (bottom) of tibialis anterior (TA) cross‐sections in control or CLP/DCS mice that received vehicle or BaCl 2 (injured) injections. Green = laminin, Blue = DAPI. Images are ×20 with scale bar = 200 μm. ( C ) Pax7 staining (top) and quantification of Pax7‐postive nuclei per myofibre of TA cross‐sections from vehicle or injured control and CLP/DCS mice. Purple = haematoxylin (nuclei), Red = Pax7. Images are ×40 with scale bar = 50 μm. All images are representative. B = nonlinear fit pairwise comparisons, see Table S6 for all P values from all comparisons. C = One‐way ANOVA with Bonferroni multiple comparisons (BMC). Significance is * P < 0.05, ** P < 0.01, *** P < 0.001. N = 3–6. Each n = one mouse.

Article Snippet: Permeabilization was performed using 0.5% Triton X‐100 (Sigma) in PBS for 5 min at room temperature (RT), then washed ×2 in immunofluorescence buffer (IFB: 3% bovine serum albumin [BSA, GoldBio], 0.2% Triton X‐100 (Sigma), 0.2% Tween‐20 [Sigma], in PBS).

Techniques: Control, Ligation, Immunofluorescence, Staining