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. 2019 Nov 19;5(11):e02864.
doi: 10.1016/j.heliyon.2019.e02864. eCollection 2019 Nov.

An experimental model of asthma in rats using ovalbumin and lipopolysaccharide allergens

Vandana R Thakur  1 , Vikas Khuman  1 , Jayesh V Beladiya  1 , Kiranj K Chaudagar  1 , Anita A Mehta  1
Affiliations

An experimental model of asthma in rats using ovalbumin and lipopolysaccharide allergens

Vandana R Thakur et al. Heliyon. .

Abstract

Asthma is chronic and multi-factorial inflammatory disease hence single allergen induced asthma in an animal is not identical to clinical asthma. Therefore, we developed a novel experimental model of asthma in rats using ovalbumin (OVA) and lipopolysaccharide (LPS) allergens. Rats were divided into four groups; normal (NC), OVA, LPS, and OVA-LPS treated. Rats were sensitized with OVA (100 μg/kg, adsorbed in 100 mg/mL aluminum hydroxide, i.p.), LPS (10 μg/kg, i.p.) and both (OVA-LPS) on 7th, 14th, 21st days and was followed by challenge with OVA (1%w/v), LPS (1%w/v), OVA (0.5%w/v) and LPS (0.5%w/v) for 30 min thrice/week for three weeks in the OVA, LPS and OVA-LPS groups, respectively. On 41 day, lung function parameters (respiration rate, tidal volume, and airflow rate), total and differential leukocytes count in the blood as well as BALf and inflammatory cytokines (IL-4, IL-5, and IL-13) in serum were measured. Histology of lungs was performed. The results suggested that the tidal volume and airflow rate were significantly decreased while respiration rate, total and differential leukocytes count in blood as well as BALf and serum cytokines level were significantly increased in the OVA-LPS as compared to NC, OVA, and LPS. In conclusion, the combination of OVA and LPS induced phenotypes of severe asthma with eosinophilic, neutrophilic and lymphocytic inflammation.

Keywords: Allergology; Asthma; Eosinophilia; Immune response; Immunology; Inflammation; Lipopolysaccharide; Neutrophilia; Ovalbumin; Pathophysiology.

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Figures

Fig. 1
Fig. 1
Schedule for sensitization and challenging protocol for the asthma induction in rats.
Fig. 2
Fig. 2
Effects of allergens sensitization and challenge on (a) respiration rate, (b) tidal volume and (c) airflow rate in normal and asthmatic rats. Normal Control (NC), Ovalbumin Control (OC), Lipopolysaccharide Control (LC), Ova-LPS Control (OLC). **p < 0.01 and ***p < 0.001 vs. NC. #p < 0.05, and ##p < 0.01 vs. OVA. &p < 0.05 and &&p < 0.01 vs. LPS.
Fig. 3
Fig. 3
Effects after 24 h of last allergen challenge on blood (a) total leukocytes count, (b) neutrophils, (c) eosinophils, (d) lymphocytes and (e) monocytes in normal and asthmatic rats. Normal Control (NC), Ovalbumin Control (OC), Lipopolysaccharide Control (LC), Ova-LPS Control (OLC). *p < 0.05, **p < 0.01 and ***p < 0.001vs. NC. #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. OVA. &p < 0.05, &&p < 0.01 and &&&p < 0.001 vs. LPS.
Fig. 4
Fig. 4
Effects after 24 h of last allergen challenge on bronchoalveolar lavage (BALf) (a) total leukocytes count, (b) neutrophils, (c) eosinophils, (d) lymphocytes and (e) monocytes in normal and asthmatic rats. Normal Control (NC), Ovalbumin Control (OC), Lipopolysaccharide Control (LC), Ova-LPS Control (OLC). *p < 0.05, **p < 0.01 and ***p < 0.001vs. NC. #p < 0.05, ##p < 0.01 and ###p < 0.001 vs. OVA. &p < 0.05 and &&&p < 0.001 vs. LPS.
Fig. 5
Fig. 5
Effects after 24 h of last allergen challenge on serum (a) IL-4, (b) IL-5 and (c) IL-13 in normal and asthmatic rats. Normal Control (NC), Ovalbumin Control (OC), Lipopolysaccharide Control (LC), Ova-LPS Control (OLC). ***p < 0.001 and **p < 0.01 vs. NC. #p < 0.05 and ###p < 0.001 vs. OVA. &p < 0.05 and &&&p < 0.001 vs. LPS.
Fig. 6
Fig. 6
Microscopic structure of H&E stained lung (transverse section) in normal and asthmatic rats. (100X original magnification) (a) Normal Control (NC), (b) Ovalbumin Control (OC), (c) Lipopolysaccharide Control (LC), (d) Ova-LPS Control (OLC).
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