Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

Article Title: Changes in intestinal homeostasis and immunity in a cigarette smoke- and LPS-induced murine model for COPD: the lung-gut axis

doi: 10.1152/ajplung.00486.2021

Figure Lengend Snippet: Inflammatory markers in serum. Mice were exposed to air or cigarette smoke for 72 days, except on days 42 , 52 , and 62 . On these days mice were treated with saline or lipopolysaccharide (LPS) via intratracheal instillation. Serum was collected and the concentration of inflammatory mediators was determined. C-reactive protein (CRP; A ), keratinocyte-derived chemokine (KC; B ), and VEGF-A ( C ) levels were detected by using an ELISA and Milliplex Luminex assay kit, respectively. Values are expressed as means ± SE. * P < 0.05; cigarette smoke (CS) group compared with air group. Correlations between KC levels in bronchoalveolar lavage (BAL) fluid and KC levels in serum ( D ) were analyzed by using Spearman correlation. Air + vehicle: n = 12 animals; air + LPS: n = 13 animals; CS + vehicle: n = 12 animals; CS + LPS: n = 13 animals.

Article Snippet: C-reactive protein (CRP) levels in the BAL fluid and serum were measured by ELISA (Mouse CRP ELISA kit, R&D Systems) according to the manufacturer’s instructions and using a microplate reader (Glomax Discover, Promega).

Techniques: Saline, Concentration Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay, Luminex

Journal: NPJ Vaccines

Article Title: Intranasal delivery of mRNA expressing newly identified Acinetobacter baumannii antigens protects against bacterial lung disease

doi: 10.1038/s41541-025-01202-0

Figure Lengend Snippet: Δ TolR::AmpR E. coli possessing each plasmid to express either OXA-23 or PAL were grown in liquid culture with 400 mM IPTG to induce protein expression over 24 h. The rOMVs were separated by SDS-PAGE for Western blot using serum from mice immunised subcutaneously with BAL_276 derived OMVs to show expression of OXA-23 ( A ) and PAL ( B ) proteins on E. coli derived rOMVs. BALB/c mice were vaccinated subcutaneously with 500 FU (~5 μg) E. coli derived rOMVs expressing PAL or OXA-23 individually or in combination in a prime boost approach with 21-day intervals. Mice were challenged intranasally with A. baumannii BAL_276 21 days after the boost and culled 24 h later ( C ). Whole bacterial-cell specific IgM ( D ) and IgG ( E ) titres were measured by ELISA. Weight loss ( F ), IL-6 level in the lungs ( G ) and bacterial loads in nasal lavage ( H ), BAL fluid ( I ), lung homogenate ( J ) and spleen homogenate ( K ) were measured 24 h after challenge. One-way ANOVAs with Tukey tests were performed to determine significant differences between groups where ** shows p < 0.01, * shows p < 0.05; n = 5 per group.

Article Snippet: To measure IL-6 levels in the BAL fluid and lungs, the R&D systems mouse IL-6 DuoSet ELISA kit was used (DY406; LOD 15.6 pg/ml).

Techniques: Plasmid Preparation, Expressing, SDS Page, Western Blot, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: NPJ Vaccines

Article Title: Intranasal delivery of mRNA expressing newly identified Acinetobacter baumannii antigens protects against bacterial lung disease

doi: 10.1038/s41541-025-01202-0

Figure Lengend Snippet: HEK293T cells were transfected with mRNA encoding either OXA-23 ( A ) or PAL ( B ) and left for 24 h. Cell lysates and cell supernatants were separated by SDS-PAGE for Western blot with serum from mice previously immunised BAL_276 derived OMVs. BALB/c mice were vaccinated intramuscularly with 5 μg LNP formulated mRNA encoding PAL or OXA-23 in a prime boost approach with 21-day intervals. Mice were challenged intranasally with A. baumannii BAL_276 21 days after the boost and culled 24 h later. Their whole bacterial cell specific IgG ( C ) and IgM ( C ) titres were measured by ELISA. Weight loss ( E ), IL-6 level in the lungs ( F ) and bacterial loads in BAL fluid ( G ), nasal lavage ( H ), lung homogenate ( I ) and spleen homogenate ( J ) were measured 24 h after challenge. One-way ANOVAs with Tukey tests were performed to determine significant differences between groups where **** shows p < 0.0001, *** shows p < 0.001, ** shows p < 0.01, * shows p < 0.05; n = 5 per group.

Article Snippet: To measure IL-6 levels in the BAL fluid and lungs, the R&D systems mouse IL-6 DuoSet ELISA kit was used (DY406; LOD 15.6 pg/ml).

Techniques: Transfection, SDS Page, Western Blot, Derivative Assay, Enzyme-linked Immunosorbent Assay