Alternaria alternata extract

Manufactured by Stallergenes Greer

Sourced in United States

Alternaria alternata extract is a laboratory product used for research and testing purposes. It is a standardized extract derived from the fungus Alternaria alternata. The core function of this extract is to provide a consistent and reliable source of Alternaria alternata allergens for various applications, such as diagnostic testing and research studies.

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Lab products found in correlation

Rmil 33, by BioLegend (3 mentions) Mouse ril 33, by R&D Systems (2 mentions) Crude peanut extract, by Stallergenes Greer (2 mentions) Pd 1 blocking antibody, by BioXCell (1 mentions) 2 deoxyglucose 2 dg, by Merck Group (1 mentions) Ova protein, by Merck Group (1 mentions) Recombinant il 13, by BioLegend (1 mentions) Recombinant il 1α, by R&D Systems (1 mentions) Limulus amebocyte lysate assay, by Lonza (1 mentions) Ril 33, by Thermo Fisher Scientific (1 mentions) Neuraminidase, by Merck Group (1 mentions)

12 protocols using alternaria alternata extract

Airway Hyperresponsiveness Measurement in Mice

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The mice were intranasally challenged via rmIL-33 (BioLegend, San Diego, Calif) or Alternaria alternata extracts (Greer Laboratories, Lenoir, North Carolina), as shown in the experimental schemes. For CD200R engagement, mouse CD200-Fc (3355-CD-050; R&D Systems), human CD200-Fc (2724-CD-050; R&D Systems) and the corresponding IgG1 isotypes (110-HG-100; R&D Systems) were used. Lung function was measured by direct measurement of lung resistance and dynamic compliance in anesthetized tracheostomized mice, in which mice were mechanically ventilated via the FinePointe RC system (Buxco Research Systems, Wilmington, NC), and sequentially challenged with aerosolized increasing doses of methacholine^{38 (link)}. Maximum lung resistance and minimum compliance values were recorded during a 3-min period after each methacholine challenge. AHR data were analyzed by repeated measurements of a general linear model.

Shafiei-Jahani P., Helou D.G., Hurrell B.P., Howard E., Quach C., Painter J.D., Galle-Treger L., Li M., Loh Y.H, & Akbari O. (2021). CD200–CD200R immune checkpoint engagement regulates ILC2 effector function and ameliorates lung inflammation in asthma. Nature Communications, 12, 2526.

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Assessing Airway Hyperresponsiveness in Mice

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Mice were intranasally (i.n.) challenged for 3 days with mouse rm-IL-33 (0.5 μg per mouse in 50 μL, BioLegend) to induce AHR or for 5 consecutive days with 100 μg of Alternaria alternata extracts (Greer Laboratories). Control mice were challenged with PBS and lung function was evaluated on the day following the last challenge. In some experiments, AHR was induced in Rag2^−/−Il2rg^−/− mice after an intravenous adoptive transfer of total pulmonary ILC2s (50 × 10³) sorted from IL-33-challenged WT and LAIR-1 KO mice. Lung function was assessed by direct measurement of lung resistance and dynamic compliance (cDyn) in restrained, tracheostomized, and mechanically ventilated mice using the FinePointe RC system (Buxco Research Systems) under general anesthesia. Mice were sequentially challenged with aerosolized PBS (baseline), followed by increasing doses of methacholine ranging from 5 to 40 mg/mL. Maximum lung resistance and minimum compliance values were recorded during a 3-min period after each methacholine challenge. AHR data were analyzed by repeated measurements of a general linear model.

Helou D.G., Shafiei-Jahani P., Hurrell B.P., Painter J.D., Quach C., Howard E, & Akbari O. (2021). LAIR-1 acts as an immune checkpoint on activated ILC2s and regulates the induction of airway hyperreactivity. The Journal of allergy and clinical immunology, 149(1), 223-236.e6.

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Intranasal IL-33 Induces AHR in Mice

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Mice were sensitized intranasally for 3 days with carrier-free recombinant mouse (rm)-IL-33 (0.5 µg per mouse in 50 µL, BioLegend) to induce AHR or with PBS, as the control. In some experiments, mice were challenged for 4 consecutive days with 100 μg of Alternaria alternata extracts (Greer Laboratories). To block glycolysis in vivo, mice were injected intraperitonially on 2 consecutive days with 500 mg kg⁻¹ of 2-Deoxyglucose (2-DG) (Sigma-Aldrich) or with PBS (vehicle). For the neutralization models, Rag2^−/− mice received an intraperitoneal injection of PD-1 blocking antibody (500 μg per mouse, clone 29 F.1A12; BioXcell) or a rat IgG2aκ isotype control 1 day before rm-IL-33 or Alternaria intranasal administration.
On day 4, lung function was evaluated by direct measurement of lung resistance and dynamic compliance (cDyn) in restrained, tracheostomized, and mechanically ventilated mice using the FinePointe RC system (Buxco Research Systems) under general anesthesia. Mice were sequentially challenged with aerosolized PBS (baseline), followed by increasing doses of methacholine ranging from 5 to 40 mg mL⁻¹. Maximum lung resistance and minimum compliance values were recorded during a 3-min period after each methacholine challenge. AHR data were analyzed by repeated measurements of a general linear model.

Helou D.G., Shafiei-Jahani P., Lo R., Howard E., Hurrell B.P., Galle-Treger L., Painter J.D., Lewis G., Soroosh P., Sharpe A.H, & Akbari O. (2020). PD-1 pathway regulates ILC2 metabolism and PD-1 agonist treatment ameliorates airway hyperreactivity. Nature Communications, 11, 3998.

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Intranasal Allergen Challenge in Pla2g5-Null Mice

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C57/BL6 Wt and Pla2g5-null mice^{48 (link), 49 (link)} (9–12 wk-old males) received 25 μg of Alternaria alternata extract (Greer Laboratories, Lenoir, NC) in 20 uL of PBS or PBS alone intranasally (i.n.) on days 0,3,6 and 9 and euthanized 18h later^{1 (link)} or a single dose of 100 μg and were euthanized after 1h or 3h². Alternatively, Wt and Pla2g5-null naïve mice were given mouse rIL-33 (R&D Systems, Minneapolis, MN) i.n. 100 ng/dose on days 0,3,6 and 9 with or without LA (132 nM)^{28 (link)} OA (106 nM) or AA (99 nM), and mice were euthanized 18h after the last dose. All animal experiments were approved by the Animal Care and Use Committee of the Dana-Farber Cancer Institute (Boston, MA).

Yamaguchi M., Samuchiwal S.K., Quehenberger O., Boyce J.A, & Balestrieri B. (2017). Macrophages regulate lung ILC2 activation via Pla2g5-dependent mechanisms. Mucosal immunology, 11(3), 615-626.

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AAV-Mediated Prevention and Treatment of Allergic Inflammation

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For the FA prevention studies, 6-week-old female FT^+/− mice were IV injected with 1 × 10¹¹ vg per mouse of TTR-OVA, EF1α-OVA, or ApoE-hAAT-FIX vectors in a total volume of 200 μL in sterile PBS. Mice were bled from the retro-orbital plexus 4 weeks after AAV injection under anesthesia using heparinized capillary tubes. Then, mice were sensitized based on Walker et al.²⁶ (link) with 20 μg of Alternaria alternata extract (Greer Laboratories, Lenoir, NC, EEUU) followed by 200 μg of OVA protein (Sigma, San Luis, MI, EEUU). This procedure was repeated on days 0, 3, 6, 9, 12, 14, 16, and 18. FT^+/− mice were bled as previously described on day 19 and challenged through IP injection of 1 mg of OVA.
For the FA treatment studies, the same AAV vectors were injected in mice previously sensitized as described above and after 4 weeks, mice were bled and challenged.

Gonzalez-Visiedo M., Li X., Munoz-Melero M., Kulis M.D., Daniell H, & Markusic D.M. (2022). Single-dose AAV vector gene immunotherapy to treat food allergy. Molecular Therapy. Methods & Clinical Development, 26, 309-322.

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Peanut Allergy Protein Extraction

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Peanut flour was purchased from the Golden Peanut Company (Alpharetta, Ga), endotoxin was undetectable (<0.5 EU/mg flour) as previously described (7 (link)). Crude peanut extract (7 (link)) and Alternaria alternata extract (19 (link)) were purchased from Greer Laboratories (Lenoir, NC). Recombinant IL-1α was purchased from R&D Systems (Minneapolis, MN), and recombinant IL-13 was purchased from Biolegend (San Diego, Ca).

Krempski J.W., Kobayashi T., Iijima K., McKenzie A.N, & Kita H. (2020). Group 2 Innate Lymphoid Cells Promote Development of T follicular Helper (Tfh) Cells and Initiate Allergic Sensitization to Peanuts. Journal of immunology (Baltimore, Md. : 1950), 204(12), 3086-3096.

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Peanut Flour Protein Extraction Protocol

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Peanut flour (14.4% protein) was purchased from the Golden Peanut Company (Alpharetta, GA, USA) as a bulk raw material; endotoxin was undetectable (<0.5 EU/mg flour) in the product by Limulus Amebocyte Lysate assay (Lonza, Walkersville, MD, USA). Crude peanut extract (70.2% protein) and Alternaria alternata extract (20.0% protein) were purchased from Greer Laboratories (Lenoir, NC, USA). Corn flour and rice flour were purchased from Bob’s Red Mill Natural Foods (Milwaukie, OR, USA).

Dolence J.J., Kobayashi T., Iijima K., Krempski J., Drake L.Y., Dent A.L, & Kita H. (2017). Airway exposure initiates peanut allergy by involving the IL-1 pathway and T follicular helper cells in mice. The Journal of allergy and clinical immunology, 142(4), 1144-1158.e8.

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Intranasal Allergen and Cytokine Challenge in Mice

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Adult female IL13-eGFP (kind gift from A.N. McKenzie, MRC Laboratory of
Molecular Biology, Cambridge) (28 (link)) were
used for experiments between 7 and 10 weeks of age. Mice were housed in
specific-pathogen-free conditions and given food and water ad libitum. All
procedures were conducted in accordance to the institutional guidelines and
under the approval of our Home Office project licence (granted under the Animals
(Scientific Procedures) Act 1986). Mice were administered 10μg
(Alternaria alternata) extract (Greer Laboratories) in
25μl of PBS intranasally 3 times a week for 1 week or PBS alone as
controls. Alternatively, carrier-free recombinant murine rIL-33 (1μg per
dose in 25μl PBS) (eBioscience) or PBS was administered 3 times a week
for 1 week. Mice were culled 24hrs after the final cytokine or allergen
dose.

Puttur F., Denney L., Gregory L.G., Vuononvirta J., Oliver R., Entwistle L.J., Walker S.A., Headley M.B., McGhee E.J., Pease J.E., Krummel M.F., Carlin L.M, & Lloyd C.M. (2019). Pulmonary environmental cues drive group 2 innate lymphoid cell dynamics in mice and humans. Science immunology, 4(36), eaav7638.

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Ormdl3-EGFP Fusion Protein for Asthma Research

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An Ormdl3-enhanced green fluorescent protein (EGFP) (N-terminal tag) gene fusion was constructed and cloned into plasmid pZac2.1 under control of the ubiquitous cytomegalovirus promoter. Adeno-associated viral vectors (AAVs) were produced by Penn Vector Core (University of Pennsylvania, Pa). On day 5 of life, neonatal mice were intranasally administered 1 × 10¹¹ genome copies of either AAV EGFP (control vector) or AAV Ormdl3-EGFP (AAV Ormdl3) with 50 mU neuraminidase (Sigma-Aldrich, Dorset, United Kingdom [UK]) in PBS. At age 8 weeks, mice received either 10 μg Alternaria alternata extract (Greer Laboratories, Lenoir, NC) or PBS via intranasal instillation, 3 times per week for 5 weeks. Mice were killed 18 hours postfinal Alternaria instillation.

Löser S., Gregory L.G., Zhang Y., Schaefer K., Walker S.A., Buckley J., Denney L., Dean C.H., Cookson W.O., Moffatt M.F, & Lloyd C.M. (2017). Pulmonary ORMDL3 is critical for induction of Alternaria-induced allergic airways disease. The Journal of Allergy and Clinical Immunology, 139(5), 1496-1507.e3.

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Intranasal Alternaria Exposure in Mice

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Ten- to 12-week-old male wild-type (WT) and Ormdl3 KO^Mer mice were bred at Taconic Biosciences (Germantown, New York, NY). WT and Ormdl3 KO^Har mice (Harwell, UK) were bred at Imperial, and neonatal mice were used for overexpression studies. All mice were housed in specific pathogen-free conditions and given food and water ad libitum. Mice were exposed to 20 μg of purified Alternaria alternata extract (Greer Laboratories) (in 25μL PBS) intranasally 3 d/wk for 5 weeks. All procedures were conducted in accordance with the Animals (Scientific Procedures) Act 1986.

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