Journal: bioRxiv

Article Title: Biofilm-Derived Curli and Z-DNA Shape Anti-DNA Antibody Responses During Salmonella Infections

doi: 10.64898/2026.03.02.708989

Figure Lengend Snippet: (A-B) 129X1/SvJ mice were intragastrically infected with STm IR715 or an isogenic non-invasive mutant STm ( invAspiB ). The serum was collected 13 weeks post-infection, diluted 1:600, and assayed by ELISA for (A) anti-Z-DNA and (B) anti-B-DNA antibodies. (C-E) 129X1/SvJ mice were fed a lithogenic (high-cholesterol) diet for 8 weeks, then switched to a normal diet for 1 week before i.p. infection with STm. (C) Serum from 3 weeks post-infection was diluted 1:250 and analyzed by ELISA for anti-dsDNA autoantibodies. (D-E) Cecal tissue was stained with antibodies against Salmonella (green) and Z-DNA (blue), EBBA Biolight 680 bacterial amyloid dye for curli (red), and DAPI for host nuclei (cyan). Representative images were taken at 63X magnification on the confocal microscope and processed via ImageJ.

Article Snippet: The following commercial antibodies were used: 5μg/mL rabbit IgG isotype control (AC042), 5μg/mL mouse IgG2b isotype control (02-6300), 5μg/mL mouse anti-Z-DNA/anti-Z-RNA antibody [Z22] (Absolute Antibodies Ab00783-3.0) (Z-DNA), 5μg/mL rabbit anti-dsDNA antibody [DSD/4054R] (Novus Biologicals NBP3-07302-100ug) (B-DNA).

Techniques: Infection, Mutagenesis, Enzyme-linked Immunosorbent Assay, Staining, Microscopy

Journal: Journal of Translational Autoimmunity

Article Title: Vascular injury derived apoptotic exosome-like vesicles trigger autoimmunity

doi: 10.1016/j.jtauto.2024.100250

Figure Lengend Snippet: Fig. 3. ApoExos trigger the production of autoantibodies. (a) ELISA was performed on the sera from ApoExos- or vehicle-injected WT mice (n = 5), and a significant increase of circulating total IgG was observed (p=0.0481). (b) Microarray experiments of multiple autoantigens showed an SLE-associated autoantibody profile in mice injected with ApoExos, but not in those injected with vehicle. Autoantibodies detected were IgG. Each row of the heatmap represents one animal for a total of 12 ApoExos-injected WT mice and 10 vehicle-injected WT mice. The results were compared by ANOVA and minimal q value of represented data was 0,05. Blue and yellow are low and high level respectively. ELISA of sera from ApoExos- or vehicle-injected mice confirmed the increased levels of SLE-associated autoantibodies (i.e., (c) anti-LG3 (p=0.0019), (d) antinuclear antibodies (ANA) (p=0.0009), (e) anti-dsDNA (p=0.0212), (f) anti-Sm/nRNP (p=0.0124), (g) anti-Sm (p=0.0008), (h) anti- SSA (Ro) (p=0.0195), (i) anti-SSB (La) (p=0.0002), (j) anti-cardiolipin (CL) (p=0.0001) and (k) anti-β2GPI (p<0.0001)), while autoantibodies related to autoim munity in transplantation were not affected (i.e., (l) anti-AT1R, (m) anti-vimentin and (n) anti-fibronectin). Statistical significance was assessed by Student’s t-test. ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; ns not significant. Results are expressed as indicated: AU, Arbitrary Units; OD, Optical Density. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Antinuclear antibodies (ANA) levels were assessed using ANA mouse bioassay kits (USBiologicals, Swampscott, MA, USA); total IgG levels were assessed with mouse IgG Total Ready-SET-Go kits (Affimetrix, Vienna, Austria); anti-double strand DNA (anti-dsDNA) levels were assessed with anti-dsDNA mouse ELISA kits (BioVendor, Neuss, Deutschland); and the levels of angiotensin 1 receptor antibodies (antiAT1R) were assessed with BioAssayTM ELISA Kits, mouse (USBiological, Salem, MA, USA), all in accordance with the manufacturers’ instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Microarray, Immunopeptidomics, Transplantation Assay

Journal: JCI Insight

Article Title: Type II alveolar epithelial cell–specific loss of RhoA exacerbates allergic airway inflammation through SLC26A4

doi: 10.1172/jci.insight.148147

Figure Lengend Snippet: ( A ) Schematic representation of HBECs expressing either a constitutively active RHOA (RHOA-L63) or dominant-negative (RHOA-N19) for SLC26A4 regulation by RhoA and subsequently TGF-β1 secretion. ( B ) qPCR analysis of relative SLC26A4 expression in HBECs expressing either RHOA-L63 or RHOA-N19 ( n = 4). ( C ) Level of TGF-β1 in supernatants of RHOA-L63– or RHOA-N19–transfected HBECs treated with CRE ( n = 4). ( D ) Representative immunofluorescence images of Slc26a4 (red) expression in lung tissue AT2 cells (Sftpc, green) of CRE-induced mouse model of asthma with RhoA fl/fl or Sftpc-cre;RhoA fl/fl mice. ( E ) Quantification of relative Slc26a4 expression in E ( n = 6). Data represent mean ± SEM. Group comparisons were made using 2-way ANOVA ( B , C , and E ). * P < 0.05, ** P < 0.01, and *** P < 0.001. Total original magnification, ×20.

Article Snippet: Human SLA26A4 in serum was detected by using Human SLC26A4 ELISA kit (orb562738, Biorbyt).

Techniques: Expressing, Dominant Negative Mutation, Transfection, Immunofluorescence

Journal: JCI Insight

Article Title: Type II alveolar epithelial cell–specific loss of RhoA exacerbates allergic airway inflammation through SLC26A4

doi: 10.1172/jci.insight.148147

Figure Lengend Snippet: ( A ) Representative immunofluorescence images of SLC26A4 (red) expression in lung tissue AT2 cells (SFTPC, green) of asthmatics and healthy controls. ( B ) Quantitative analysis of SLC26A4 expression in A ( n = 5). ( C ) Serum levels of SLC26A4 in healthy subjects ( n = 18) and patients with mild ( n = 31) and severe ( n = 24) asthma. ( D ) Correlation between serum levels of SLC26A4 (pendrin) and FEV 1 % among asthmatics ( n = 55). ( E and F ) Correlation between serum levels of SLC26A4 (pendrin) and FEV 1 % among the mild ( E , n = 31) and severe ( F , n = 23) asthmatics, respectively. ( G ) Representative images of SLC26A4 expression in HBECs after CRE treatment. ( H ) Quantification of SLC26A4 + staining in G ( n = 5). Data represent mean ± SEM. Group comparisons were made using 2-tailed Student’s t test ( B and H ) and 2-way ANOVA ( C ). * P < 0.05, ** P < 0.01, and *** P < 0.001. Scale bars: 25 μm.

Article Snippet: Human SLA26A4 in serum was detected by using Human SLC26A4 ELISA kit (orb562738, Biorbyt).

Techniques: Immunofluorescence, Expressing, Staining

Journal: JCI Insight

Article Title: Type II alveolar epithelial cell–specific loss of RhoA exacerbates allergic airway inflammation through SLC26A4

doi: 10.1172/jci.insight.148147

Figure Lengend Snippet: ( A ) Epithelial cell–derived cytokines in the BALF of NPPB treated mice ( n = 8–10). ( B ) Epithelial cell–derived cytokines in supernatants of CRE-treated HBECs in the presence or absence of NPPB treatment ( n = 4–6). ( C ) Epithelial cell–derived cytokines in supernatants of CRE-treated HBECs with or without SLC26A4 knockdown ( n = 4). Data represent mean ± SEM of 2 independent experiments. Group comparisons were made using 2-way ANOVA ( A – C ). * P <0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Human SLA26A4 in serum was detected by using Human SLC26A4 ELISA kit (orb562738, Biorbyt).

Techniques: Derivative Assay, Knockdown