4-6 weeks old male Wistar rats weighing 80-120 g at study initiation, were obtained/purchased from the Animal House of Ahvaz Jondishapour University of Medical Sciences (AJUMS), Ahvaz, Iran. The rats were housed in an animal room maintained at 23 ± 3 °C and a relative humidity of 30-70 % with the altering light/dark cycle of 12h during the experiment and for at least, one week prior to sensitization period (for acclimatization purposes). The animals were housed in stainless-steel wire cages in groups of 5 and had free access to PN-free standard laboratory rodent chow and water.
All protocols involving the examined animals, were orchestrated in conformity with the instructions for Laboratory Animal Experiments in AJUMS Animal Research & Care Center.
The reagents used in this research, were purchased from the sources enclosed in parentheses:
Alum [Al(OH)3] (Alhydrogel 2.0%, Serva Chemical Co., U.S.A), Cholera toxin (C-3012, Sigma Chemical Co., St. Louis, Mo, U.S.A), DMSO (Merck Chemical Co., Germany), Evan's blue dye (Merck Chemical Co., Germany), Rat histamine kit (LDN Chemical Co., Germany), K3-EDTA (Sigma Chemical Co., St. Louis, Mo, U.S.A), PBS (Merck Chemical Co., Germany), Rat Total IgE kit (ICL Chemical Co, U.S.A).
Unshelled/incrusted crude PNs were supplied from Safee-Abad Tree-plantar Research Station in the town of Dezfool, Khuzestan Province, Iran.
In the present study, PN proteins were used as allergens which were extracted from fresh crude PNs as follows, briefly:
PN bodies were pulverized by a mill and the resulted paste was defatted by n-Hexane (1:3 v/v, 3 times). Subsequent to separation process, residues were dried out and deodorized via gentle he...
... middle of paper ...
...t response was defined as a wheal reaction showing up as a blue area/circle measuring greater than 5 mm in diameter when read at 20 min post id PN-challenge.
Detection of Vascular Leakage
Five min before intra-peritoneal PN challenge-dose adminstration, 7 rats from each group received 200 μL of Evan's blue dye (5 mg/mL PBS) by tail vein injection. Footpads (& paws) of mice were examined for signs of vascular leakage (visible blue color) 40-45 min after ip administering of 200 μg of the filter-sterilized CPE.
Data were analyzed using the SigmaStat statistical software package (SPSS, Chicago, Ill, Version 15). Statistical differences between two groups were analyzed by 1-way ANOVA. A probability value of less than 0.05 was considered statistically, significant.
Adjuvant, Anaphylactic Parameters, Peanut Allergy, Wistar-strain Model
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