Carboxymethyl cellulose (CMC) sodium salt, DEXA-water soluble, OVA, trypan blue, aluminum hydroxide gel, and Al(OH)3 were obtained from Sigma-Aldrich (St. Louis, MO, USA). Isoflurane was obtained from Hana Pharm. Co. (Hwaseong, Korea). IL-4, IL-5, and an OVA-specific IgE (OVA-sIgE) ELISA kit were obtained from MyBioSource (San Diego, CA, USA). TRIzol reagent was obtained from Invitrogen (Carlsbad, CA, USA).
Aluminum hydroxide gel
Manufactured by Merck Group
Sourced in United States
Aluminum hydroxide gel is a laboratory product used as an adsorbent material. It is a fine, white, gelatinous substance composed of aluminum hydroxide. The primary function of aluminum hydroxide gel is to adsorb and remove impurities or unwanted substances from solutions.
Automatically generated - may contain errors
Lab products found in correlation
Ovalbumin (ova), by Merck Group (7 mentions)
Powerlab 800, by ADInstruments (2 mentions)
Trypan blue, by Merck Group (1 mentions)
Carboxymethyl cellulose cmc sodium salt, by Merck Group (1 mentions)
Al oh 3, by Merck Group (1 mentions)
Interleukin 4 (il 4), by MyBioSource (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Anti il 17a apc, by BioLegend (1 mentions)
Bicinchoninic acid (bca), by Beyotime (1 mentions)
Anti il 4 pe, by BD (1 mentions)
Anti gr1, by BioLegend (1 mentions)
Max 1320, by Buxco Electronics (1 mentions)
Pan cytokeratin, by Santa Cruz Biotechnology (1 mentions)
Fixable viability stain 510 antibody, by BD (1 mentions)
Mbd2 antibody, by Abcam (1 mentions)
Anti ecp, by Biorbyt (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Application suite v4, by Leica (1 mentions)
Enzyme immunoassay, by Cayman Chemical (1 mentions)
24 protocols using aluminum hydroxide gel
1
OVA-Induced Allergic Airway Model
2
Asthma Mouse Model Reagents
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GREER laboratories supplied HDM; OVA, LPS, and aluminum hydroxide gel from Sigma; mice spirometer (MAX 1320) from Buxco®, USA; Beijing Zhongshidichuang Science and Technology Development Co. Ltd. supplied animal asthma inducing instrument (YLS-8A); centrifuge (5810 R) from Eppendorf®, Germany; image analyzer (Leica Application Suite V4) from Leica, Germany; and Olympus, Japan (Leica®) supplied optical microscope (DMI3000B); 150-mesh cell sieve (Biosharp, BS-100-XBS, China); bronchial epithelial growth medium (Procell, CM-M007, China); DAPI and cytokeratin specific monoclonal antibody (pan-Cytokeratin, Santa Cruz, sc-8018, USA); leukocyte activation cocktail (550583, BD Biosciences, USA); cell viability marker (Fixable Viability Stain 510 antibody, BD Pharmingen); PE-anti-IL-4 (BD Pharmingen); APC-anti-IL-17A (Biolegend); Lipofectamine 3000 (Invitrogen, USA); bicinchoninic acid (BCA), Beyotime, Shanghai, China; DHT (B8214) and 17β-estradiol (C4348) APExBIO, USA; MBD2 antibody (Abcam, Cambridge, USA); eosinophil antibody (anti-ECP, Biorbyt, Cambridge, UK); neutrophil antibody (anti-Gr-1, Biolegend, San Diego, USA); GATA3 (10417-1-AP) and β actin (60008-1-Ig) Proteintech, USA; RORγt (EPR20006) abcam, USA.
3
Tolerance Induction to Bt Extract
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Four groups of A strain mice (n=7–8) were used: 1) naïve controls; 2) sensitized with BtE; 3) transplanted with one dose of TolDCs and sensitized with BtE; and 4) transplanted with 2 doses of TolDCs, sensitized with BtE. To induce tolerance to BtE, mice were injected intraperitoneally with 106 TolDCs, in 100 µL of saline 10 and/or 5 days before the first sensitizing exposure to BtE. Mice were sensitized with intraperitoneal injections of BtE containing 100 µg of protein adsorbed to 1.6 mg of aluminum hydroxide gel (Sigma-Aldrich), and a booster was injected 14 days later. Challenge was induced intranasally with 4 applications of a solution containing 10 µg of BtE in 25µL of saline starting 7 days after booster at 1-day intervals. The naïve control group received only saline injections. Mice were euthanized 24 hours after the last challenge.
4
Formalin-Inactivated Respiratory Syncytial Virus Preparation
5
OVA-Induced Mouse Airway Allergy Model
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Mice were immunized and challenged with OVA (Sigma‐Aldrich, St Louis, MO) according to our previously described method18 , 25 (Fig. 1A ). Briefly, mice were intraperitoneally injected (50 µg/mouse) with OVA in phosphate‐buffered saline (PBS) mixed with an equal volume of 4 mg/mL aluminum hydroxide gel (Sigma‐Aldrich) as an adjuvant in a total volume of 1 mL at the indicated time‐points on days 1, 7, and 14, respectively. This was followed by intranasal administration of 3% OVA solution in PBS into the bilateral nasal cavity, either in the morning (ZT4) or evening (ZT16), daily for 12 consecutive days from days 21 to 32. PBS alone was administered as a control (Fig. 1B ).
6
Bronchial Reactivity and LTC4 Levels in Allergic Mice
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BALB/c mice were sensitized to OVA, and bronchial reactivity and pulmonary LTC4 levels were measured62 (link). On days 0 and 7, α-T-13′-COOH (4a ; 10 mg/kg, i.p.) or vehicle (4% DMSO in saline; 0.5 ml) were administered 30 min before injection of OVA (100 µg; Sigma Aldrich) which was adsorbed to aluminum hydroxide gel (3.3 mg; Sigma Aldrich). Mice were sacrificed on day 21, and bronchi were collected. Bronchial rings of 1–2 mm length were cut and placed in organ baths mounted to isometric force transducers (Type 7006, AD Instruments, Ugo Basile, Comerio, Italy) and connected to a Powerlab 800 (AD Instruments). Rings were initially stretched until a resting tension of 0.5 g was reached and allowed to equilibrate for at least 30 min. In each experiment, bronchial rings were challenged with carbachol (10−6 M) until the response was reproducible. Bronchial reactivity was then assessed from a cumulative concentration–response curve to carbachol (1 × 10−8 − 3 × 10−5 M). To determine pulmonary LTC4 levels, lungs were isolated and homogenized in PBS pH 7.4 (100 mg tissue/ml) prior to detection of LTC4 by enzyme immunoassay (Cayman Chemicals).
7
Ovalbumin-Induced Airway Inflammation Model
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The mice were divided into 2 groups: ovalbumin intratracheally (i.t.)-challenged mice (OVA group) and the control group. The OVA group was actively sensitized with 3 intraperitoneal (i.p.) injections of 100-µg of ovalbumin (OVA, grade V, Sigma-Aldrich, St. Louis, MO, USA) emulsified in 1.3 mg of aluminum hydroxide gel (Sigma-Aldrich) in a total volume of 0.4 mL on days 1, 7, and 14. Fourteen days after the completion of the sensitization, an i.t. challenge was performed 3 times as shown in Fig. 1 . For each challenge, the mice were anesthetized with pentobarbital sodium (90 mg/kg administered i.p.) and their limbs and cheeks were tightly attached to an operation table in a supine position; a 5-mm midline cervical incision was made to expose the trachea; the table was then positioned at a 70℃-80℃ angle, and 200 µg of OVA solved in 25-µL of warm (37℃) sterile normal saline was administered. The cervical incision was stitched with a 5.0 silk suture, and the mice were held in an erect position for 40-60 minutes before being returned to their cages. The animals recovered rapidly after surgery. This procedure was performed 3 times in a biological safety cabinet (Labculture A2; Esco Micro Pte Ltd., Singapore). The control group underwent the same procedure but received saline solution instead of OVA.
8
Oral Tolerance Induction in Mice
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BALB/c mice, 6 wk of age, were purchased from Orient Inc. (Raon Bio, Korea). Induction of tolerance followed the protocol of Kim et al. (2013) (link) with minor modification. Briefly, either native or irradiated OVA (25 mg in PBS) was administrated to mice, intragastrically, in 0.5 ml using a ball-tipping feeding needle. On days 5 and 12 after feeding, mice were sensitized with the mixture of 20 μg of OVA and 2.6 mg of aluminum hydroxide gel (Sigma, USA) to induce a Th2 response.
9
Ovalbumin-Induced Airway Reactivity in Mice
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BALB/c mice received 27a (p.o.) or vehicle (0.5% carboxymethylcellulose
in 10%
Tween 20; 0.5 mL) on days 0 and 7, 1 h (p.o) before being sensitized
to ovalbumin (100 μg adsorbed to 3.3 mg of aluminum hydroxide
gel, s.c., Sigma-Aldrich). Mice were sacrificed on days 1, 3, 8, 10,
or 21 to collect lung and plasma. Bronchi were cut in rings of 1 to
2 mm length, placed in organ baths, and fixed to an isometric force
transducer 7006 connected to a Powerlab 800 (AD Instruments, Ugo Basile,
Comerio, Italy). After stretching the rings to a resting tension of
0.5g and equilibration for at least 30 min, the rings
were challenged with carbachol (1 μM) until a reproducible response
was observed. To assess bronchial reactivity, the cumulative response
to carbachol (0.001 to 3.16 μM) was measured. Bronchial relaxation
was determined from the cumulative response of precontracted bronchial
tissues to salbutamol. Compound27a , LTB4,
and LTB4 isomers were analyzed in plasma and lung homogenates
by UPLC-MS/MS. Lung tissue (100 mg/mL) was homogenized in PBS pH 7.4
at 4 °C for 1–2 min using an Omni tissue homogenizer (Omni,
Kennesaw, GA).
in 10%
Tween 20; 0.5 mL) on days 0 and 7, 1 h (p.o) before being sensitized
to ovalbumin (100 μg adsorbed to 3.3 mg of aluminum hydroxide
gel, s.c., Sigma-Aldrich). Mice were sacrificed on days 1, 3, 8, 10,
or 21 to collect lung and plasma. Bronchi were cut in rings of 1 to
2 mm length, placed in organ baths, and fixed to an isometric force
transducer 7006 connected to a Powerlab 800 (AD Instruments, Ugo Basile,
Comerio, Italy). After stretching the rings to a resting tension of
0.5g and equilibration for at least 30 min, the rings
were challenged with carbachol (1 μM) until a reproducible response
was observed. To assess bronchial reactivity, the cumulative response
to carbachol (0.001 to 3.16 μM) was measured. Bronchial relaxation
was determined from the cumulative response of precontracted bronchial
tissues to salbutamol. Compound
and LTB4 isomers were analyzed in plasma and lung homogenates
by UPLC-MS/MS. Lung tissue (100 mg/mL) was homogenized in PBS pH 7.4
at 4 °C for 1–2 min using an Omni tissue homogenizer (Omni,
Kennesaw, GA).
10
Immunization with Aluminum-Adsorbed recLytB
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Three six-week-old female BALB/c mice (Charles River, Italy) were immunized subcutaneously with 50 μg of recLytB adsorbed to aluminum hydroxide. To obtain the aluminum-adsorbed antigen, recLytB (500 μg) was resuspended in 1 ml of 0.15 M NaCl and added to a solution of 2 mg ml-1 Aluminum hydroxide gel (Sigma-Aldrich, St Louis, MO, USA) in NaCl 0.15 M. The suspension was incubated in agitation at room temperature for 1 hour, and then centrifuged at 3,000 × g, at room temperature, for 5 min. Supernatant was collected and analyzed for unabsorbed protein with Bradford reagent. The pellet containing the adsorbed protein to aluminum gel (Al-LytB) was resuspendend in NaCl 0.15 M, and prepared for subcutaneous inoculation. An Al-LytB dose of 50 μg/mouse was delivered at day 0 and day 21. Blood samples were collected from temporal plexus at day 0 and day 35, and at day 37 by cardiac puncture. Total anti-LytB IgG serum was analyzed by enzyme-linked immunosorbent assay (ELISA) as previously described [20 (link)]. Sera were stored at -35°C.
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