Additional file 1 of IL-4Rα blockade reduces influenza-associated morbidity in a murine model of allergic asthma
Bibliographic record
Abstract
Additional file 1: Table E1. Primers used for qPCR assays. Figure E1. Lung Tissue Sectioning. The left lung was cut in a transverse manner to create a superior and an inferior segment. The inferior segment was embedded into paraffin, while the superior segment was further cut in a sagittal manner and embedded. Figure E2. House dust mite (HDM) sensitization delays viral clearance. Phosphate-buffered saline (PBS)-exposed or HDM-sensitized mice were infected with pandemic H1N1 (pH1N1) and sacrificed on days 0, 2, 4, 6, and 8 post-infection. (A) IFN-β protein expression was determined using ELISA on the supernatant of bronchoalveolar lavage fluid (BALF). (B) qPCR was performed on RNA from homogenized lung tissue to determine the number of viral RNA copies, and results were compared using a two-tailed Student’s t-test. Data (n=5 per group per day) were compared using a two-way ANOVA with Sidak’s multiple comparison’s test and are expressed as mean ± SEM. *P<0.05 after correction for multiple comparisons; ns: not significant. Figure E3. House dust mite (HDM) sensitization dampens the induction of interferon stimulated genes (ISGs). Phosphate-buffered saline (PBS)-exposed or HDM-sensitized mice were infected with pandemic H1N1 (pH1N1) and sacrificed on days 0, 2, 4, 6, and 8 post-infection. qPCR was performed on RNA from homogenized lung tissue to measure the gene expression of the interferon stimulated genes (A) MX1, (B) IFITM3, (C) OAS1, (D) ISG15, (E) Viperin, and (F) RIG-I. Results were normalized to expression on day 0 and were compared using a Mann-Whitney U test. Data (n=5 per group per day) are expressed as mean ± SEM. *P<0.05 after correction for multiple comparisons; **p<0.01 after correction for multiple comparisons; ns: not significant. Figure E4. House dust mite (HDM)-sensitized mice present elevated IL-33 and lower IFN-γ levels following pandemic H1N1 (pH1N1) infection. Phosphate-buffered saline (PBS)-exposed or HDM-sensitized mice were infected with pandemic H1N1 (pH1N1) and sacrificed through a time-course study. (A) IL-33 and (B) IFN-γ protein levels were measured using ELISA and multiplex assay, respectively (n=5 per group per day). Data were compared using two-way ANOVA with Sidak’s multiple comparison’s test and are expressed as mean ± SEM. *P<0.05 after correction for multiple comparisons; ** P<0.01 after correction for multiple comparisons; ns: not significant. Figure E5. Bronchoalveolar lavage fluid (BALF) cell differential analysis. BALF was obtained on day 8 post-infection from pandemic H1N1 (pH1N1)-infected mice. The percentage of (A-C) neutrophils, (D-F) lymphocytes, and (G-I) macrophages was determined in a total of 200 counted cells. (J-L) Trypan blue was used on the re-suspended BALF pellet and the absolute number of inflammatory cells in BALF was determined using a hemocytometer. Data were compared using a two-tailed Student’s t-test and are expressed as mean ± SEM. Blue/pink: intranasal phosphate-buffered saline (PBS) (n=10) / intranasal house dust mite extract (HDM) (n=9); green/orange: intranasal HDM followed by prophylactic strategy of IL-4Rα blockade (n=12) / IgG (n=12); black/red: intranasal HDM followed by therapeutic strategy of IL-4Rα blockade (n=12) / IgG (n=10). ** P<0.01; ***P<0.001; ns: not significant. Figure E6. Systemic blockade of Interleukin-4 receptor alpha (IL-4Rα) reduces IL-6 and IP-10 levels in bronchoalveolar lavage fluid (BALF). BALF was obtained on day 8 post-infection from pandemic H1N1 (pH1N1)-infected mice. (A-C) IL-6 and (D-F) IP-10 protein levels were measured in BALF by multiplex assay. Data were compared using the Mann-Whitney U test and are expressed as mean ± SEM. Blue/pink: intranasal phosphate-buffered saline (PBS) (n=10) / intranasal house dust mite extract (HDM) (n=9); green/orange: intranasal HDM followed by prophylactic strategy of IL-4Rα blockade (n=13) / IgG (n=12); black/red: intranasal HDM followed by therapeutic strategy of IL-4Rα blockade (n=12) / IgG (n=12). * P<0.05; ** P<0.01; ns: not significant. Figure E7. HDM-sensitization Induces Immune Cell Infiltration to the Airways. (A) PBS-exposed or HDM-sensitized mice were sacrificed on day 8 following pH1N1 infection or CAF exposure (n=10 except for HDM + pH1N1 n=9). Paraffin-embedded lung tissue obtained from these mice was sectioned and stained with Periodic Acid Schiff (PAS). Fractional lymphoid area was quantified and is expressed as a percentage of lymphoid tissue area per total cross sectional area. Data are expressed as mean ± SEM by one-way ANOVA. (B) PBS-exposed or HDM-sensitized nice were sacrificed through a time-course study (n=5 per group per day). The lungs were homogenized with PBS, and the supernatant was collected for protein analysis using the BCA assay). Data are expressed as mean ± SEM by a two-way ANOVA with Sidak’s multiple comparisons test. ** P<0.01; *** P<0.001.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.003 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.874 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".