Disease Pathophysiology

• Carrageenan: Polysaccharide derived from red algae

• Acute inflammation; edema, hyperalgesia, erythema

• Pro-inflammatory agents: bradykinin, histamine, tachykinins, complement and reactive oxygen, and nitrogen species

Disease Induction

• 0.5mg/kg of carrageenan in 0.1ml saline is injected into the right hind footpad using a 27Ga needle; 0.1ml saline is injected into the left footpad

• Alternatively, CFA (50µl) or methylated-BSA (100µg in 50µl) may be used

PD/Endpoint Analysis

• Footpad thickness measured with calipers

• Blood serum analysis for inflammatory markers

• IHC of footpad

Disease Pathophysiology

• Inflammatory bowel diseases (IBD) mainly comprised of Ulcerative Colitis and Crohn's Disease are complex and multifactorial disease with unknown etiology.

• To study human IBD mechanistically, number of murine models of colitis has been developed. Among various chemical induced colitis models, DSSinduced colitis model is widely used because of its simplicity and many similarities with human ulcerative colitis.

Disease Induction

• Colitis is induced by providing 2.5% DSS (36-50kDa) in the bottled drinking water of C57Bl/6 mice

• Control group is provided unaltered drinking water (8 total mice)

• Body weights and clinical signs are collected daily, as this model is known to induce body weight loss and dehydration

PD Endpoint Analysis

• Stool consistency is assessed daily, with occult blood testing:

  • • Normal stool consistency with negative hemoccult: 0

    • Soft stools with positive hemoccult: 1

    • Very soft stools with traces of blood: 2

    • Watery stools with visible rectal bleeding: 3

• H & E staining of fixed colon tissue

Disease Pathophysiology

• Acute inflammatory response characterized by erythema, scaling, and epidermal thickening

• Disease is dependent upon IL-23 and IL-17R signaling

Disease Induction

• Mice (C57Bl/6, Balb/c) receive 3.125mg Imiquimod through topical application of 5% Imiquimod cream (3M Pharmaceuticals), QD x 6-7 to the depilated back and right ear

PD/Endpoint Analysis

• Scoring for erythema, scaling and ear skin thickness

• IHC analysis of keratinocyte proliferation and differentiation

• Immune-cell, cytokine profiling

Disease Pathophysiology

• Small molecule irritant that induces immune response when bound to a carrier protein

• Typical AD-like lesions with edema, scabs, epithelial-cell hypertrophy, marked mast-cell and lymphocyte infiltration of dermis, and increased IL-4, IL-6, IL-10, IL-1β, IL-17A, IFN-γ and TNF-α expression.

Disease Induction

• DNFB (2,4-Dinitrofluourbenzene) (Sigma Aldrich) administration to Balb/c mice: 5 sensitizations at weekly intervals, AD lesions within 4-5 days after second dose

• The ear or dorsal skin of BALB/c mice was painted with 100 µL of 0.15% DNFB in vehicle [acetone/olive oil (3:1)]

PD/Endpoint Analysis

• Serum Ig levels, scratching behavior