alternaria alternata extract greer xpm1d3a2.5  (Greer Laboratories)

Journal: Cancer cell

Article Title: Fungal mycobiome drives IL-33 secretion and type 2 immunity in pancreatic cancer.

doi: 10.1016/j.ccell.2022.01.003

Figure Lengend Snippet: Figure 4. Intratumor fungi facilitate the secretion of IL-33 from PDAC cells (A) Representative IHC images of IL-33 in the spleen, normal pancreas, PanIN (KC mice), and PDAC (KPC mice) of 6-, 12-, and 24-week-old mice. Scale bars, 50 mm. (B) Fluorescence images showing nuclear expression of IL-33 (green) in PDAC cell line. DAPI (blue) was used for nuclear staining. Magnification 403. Scale bars, 75 mm. (C) Subcellular fractionation of Dox-inducible murine PDAC cell line showing IL-33 expression in cytoplasm and nucleus. Lamin A/C and b-tubulin were used as nuclear and cytoplasmic loading control, respectively. (D) The gut and intrapancreatic (n = 3, biologically independent samples) mycobiomes of PDAC tumor-bearing mice were analyzed by 18S internal transcribed space (ITS) sequencing. Heatmap shows relative abundance of the fungal genus (left) and family (right) in gut versus PDAC. (E) Fluorescence in situ hybridization (FISH) showing fungal population in normal pancreas versus PDAC. D223 fungal-specific probe was used to detect the fungal species in the normal pancreas. (F) Schematic showing strategy for fungal extract treatment followed by biochemical assays to determine IL-33 expression in cells and conditioned medium upon treatment with Alternaria alternata. (G) Western blot analysis of IL-33 in PDAC cell line treated with vehicle and fungal extract (A. alternata) for different time points (2, 3, 6, and 24 h) and shIL33 PDAC cell line. b-Actin was used as a loading control. (H) IL-33 was measured in conditioned medium using ELISA in PDAC cell line treated with A. alternata extract for different time points (2, 3, and 6 h) (n = 3). (I) Schematic showing strategy for quantification of IL-5 in flow-sorted ILC2 cells cultured with PDAC cell-conditioned medium treated earlier with the fungal extract. (J) IL-5 measured using ELISA produced from ILC2s upon treatment with fungal-extract-treated cancer-cell-conditioned medium. Data are presented as mean ± SD. p values were calculated using Student’s t test. ns, no significance. Individual p values are indicated in the panels. See also Figure S4.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER APC Rat IgG2a Biolegend Cat# 400512; RRID: AB_2814702 PE/Cyanine5 Rat IgG2a Biolegend Cat# 400510 PerCP/Cyanine5.5 Mouse IgG2b Biolegend Cat# 400338 Brilliant Violet 421 FOXP3 Biolegend Cat# 126419; RRID: AB_2565933 Brilliant Violet 785 CD8a Biolegend Cat# 100749; RRID: AB_2562610 Brilliant Violet 421(TM) anti-human FOXP3 Biolegend Cat# 320124; RRID: AB_2565972 APC/Cyanine7 anti-human CD45 Biolegend Cat# 304014; RRID: AB_314402 APC anti-human CD3 Biolegend Cat# 317318; RRID: AB_1937212 PE/Cyanine7 anti-human CD4 Biolegend Cat# 317414; RRID: AB_571959 Brilliant Violet 785(TM) anti-human CD8 Biolegend Cat# 344740; RRID: AB_2566202 PE anti-mouse/human KLRG1 (MAFA) Biolegend Cat# 138408; RRID: AB_10574313 APC anti-human Lineage Cocktail (CD3, CD14, CD16, CD19, CD20, CD56) Biolegend Cat# 348803 PE/Cyanine5 anti-human CD127 (IL7Ralpha) Biolegend Cat# 351324; RRID: AB_10915554 FITC anti-human CD25 Biolegend Cat# 302604; RRID: AB_314273 Human TruStain FcX Biolegend Cat# 422302; RRID: AB_2818986 Alexa Fluor(R) 488 anti-human FOXP3 Biolegend Cat# 320112; RRID: AB_430883 PerCP/Cyanine5.5 anti-GATA3 Biolegend Cat# 653812; RRID: AB_2563219 Mouse Anti-Human CD4 Becton Dickinson Cat# 560345; RRID: AB_1645572 CD3 Becton Dickinson Cat# 560176; RRID: AB_1645475 CD8 Becton Dickinson Cat# 560347; RRID: AB_1645581 CD127 (IL-7 Receptor a subunit) Becton Dickinson Cat# 557938; RRID: AB_2296056 Human ST2/IL-33R APC R&D Cat# FAB5231A Brilliant Violet 510 anti-human Lineage Cocktail (CD3, CD14, CD16, CD19, CD20, CD56) Biolegend Cat# 348807 PE anti-human CD25 Biolegend Cat# 302606; RRID: AB_314276 Bacterial and virus strains Alternaria alternata ATCC Cat# ATCC 36376 Malassezia globosa ATCC Cat# MYA-4889 Alternaria alternata extract Stallergenes Greer Lab Cat# XPM1D3a.5 Candida albicans extract Stallergenes Greer Lab Cat# XPLM73X1A2 (Continued on next page) e3 Cancer Cell 40, 153–167.e1–e11, February 14, 2022

Techniques: Fluorescence, Expressing, Staining, Fractionation, Control, Sequencing, In Situ Hybridization, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture, Produced

Journal: Nature Communications

Article Title: Group 2 innate lymphoid cells are a non-redundant source of interleukin-5 required for development and function of murine B1 cells

doi: 10.1038/s41467-024-54780-3

Figure Lengend Snippet: a – c Immunglobulin concentrations in the serum of Nmur1 iCre-eGFP Id2 fl/fl and littermate Id2 fl/fl mice were determined by using the LEGENDplex multiplex beads-based assay. a Total immunoglobulin concentration in steady state ( Id2 fl/fl n = 8, Nmur1 iCre-eGFP Id2 fl/fl n = 8). b Mice were infected with N. brasiliensis and serum was harvested at d11 post-infection ( Id2 fl/fl n = 8, Nmur1 iCre-eGFP Id2 fl/fl n = 9). c Inflammation was induced by intranasal administration of Alternaria alternata ( A. alternata ) extract. Sera were harvested at d7 after induction of inflammation (Untreated Id2 fl/fl n = 4, A. alternata Id2 fl/fl n = 12, A. alternata Nmur1 iCre-eGFP Id2 fl/fl n = 12). Each symbol represents data from one mouse, mean +/− SD, data are pooled from two ( a , b ) or three ( c ) independent experiments. Statistical significance was determined by two-tailed unpaired Mann Whitney test ( a – c ), * p < 0.05, ** p < 0.01. Source data, including exact p -values, are provided as a Source data file Fig. 5.

Article Snippet: For allergic asthma induction, 10 μg A. Alternata extract (Greer Laboratories) in PBS were administered intranasally on three consecutive days.

Techniques: Multiplex Assay, Concentration Assay, Infection, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Leukocyte Biology

Article Title: Administration of a bacterial lysate to the airway compartment is sufficient to inhibit allergen-induced lung eosinophilia in germ-free mice

doi: 10.1093/jleuko/qiae047

Figure Lengend Snippet: GF mice associated with healthy gut microbiota from birth (Ex-GF mice) exhibit attenuated AHR in 2 allergic asthma models. GF (black symbols) and Ex-GF (gray symbols) BALB/c mice were sensitized and challenged with OVA (A) or Alternaria alternata extracts (B). Shown are invasive measurements of total airway resistance (Rrs) from 2 independent experiments for each model. Numbers of mice/group are indicated in the figure captions. Symbols and bars denote mean and SEM, respectively. Statistical differences between dose-response curves of total airway resistance in GF and Ex-GF mice were assessed by linear mixed-effects models. ALT = Alternaria .

Article Snippet: In the Alternaria model, 7- to 8-wk-old GF and Ex-GF BALB/c mice were sensitized i.n. with A. alternata extracts (Greer Laboratories; 50 μg of dry weight in 50 μL of saline) at days 0 and 1, challenged i.n. with Alternaria extracts (25 μg of dry weight in 50 μL) at days 21 to 23, and sacrificed at day 26.

Techniques:

Journal: Journal of Leukocyte Biology

Article Title: Administration of a bacterial lysate to the airway compartment is sufficient to inhibit allergen-induced lung eosinophilia in germ-free mice

doi: 10.1093/jleuko/qiae047

Figure Lengend Snippet: Airway administration of a bacterial lysate but not early association with healthy mouse gut microbiota inhibits allergen-induced BAL eosinophilia in GF mice BAL cellularity was measured in GF (black bars) and Ex-GF (gray bars) BALB/c mice sensitized and challenged with OVA (left and center) or Alternaria alternata (right). Additional groups of mice (hatched bars) received allergen + 9 (OVA model) or 14 ( Alternaria [ALT] model) i.n. OM-85 treatments (1 mg/mouse/treatment). Shown are total numbers of BAL eosinophils (Eos)/mouse (mean and SEM) from 2 independent experiments for each model. Numbers of mice/group are indicated in the figure captions. Statistical differences between groups were assessed by an unpaired, 2-tailed t test after evaluating the normality of sample value distributions. Bolded P values remained significant after Bonferroni correction ( P < 0.016).

Article Snippet: In the Alternaria model, 7- to 8-wk-old GF and Ex-GF BALB/c mice were sensitized i.n. with A. alternata extracts (Greer Laboratories; 50 μg of dry weight in 50 μL of saline) at days 0 and 1, challenged i.n. with Alternaria extracts (25 μg of dry weight in 50 μL) at days 21 to 23, and sacrificed at day 26.

Techniques:

Journal: Science immunology

Article Title: The ChAT-acetylcholine pathway promotes group 2 innate lymphoid cell responses and anti-helminth immunity

doi: 10.1126/sciimmunol.abe3218

Figure Lengend Snippet: (A to C) Representative flow cytometry plots (A), population frequencies (B) and mean fluorescence intensities (C) of ChAT+ ILC2s in sort-purified small intestinal ILC2s in vitro cultured for 3 days in IL-2 and IL-7 with or without IL-33. (D to I) Representative flow cytometry plots (D and G), population frequencies (E and H) and numbers (F and I) of ChAT+ ILC2s in the lung (D to F) and mLNs (G to I) of PBS or IL-33-treated ChATBAC-eGFP mice, gated on total ILC2s. (J to L) Representative population frequencies of ChAT+ ILC2s in the mLNs (J), ceca (K) or lung (L) of TSLP or IL-25-treated (J), T. muris-infected (K), or Alternaria-treated (L) ChATBAC-eGFP mice, gated on total ILC2s. Data are representative of two independent experiments. n = 3 mice per group. Data are mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: For Alternaria treatment, 10 μg Alternaria alternata extract (Stallergenes Greer) in 40 μl PBS was intranasally administered for three consecutive days and analyzed one day later.

Techniques: Flow Cytometry, Fluorescence, Purification, In Vitro, Cell Culture, Infection