Characterisation of the pruritus responses and pruritic behaviours in an interleukin 31-induced canine model of pruritus.

BACKGROUND: Intravenous administration of interleukin (IL)-31 in healthy dogs has been used as a model to assess antipruritic drugs. However, there is no known in-depth characterisation of pruritic behaviours, and the repeatability of the IL-31-induced pruritus in the individual dogs is currently unknown. OBJECTIVES: To evaluate the immediate/delayed pruritus responses and the pruritic behaviours observed in the IL-31-induced pruritic model in healthy dogs after repeated IL-31 injections. ANIMALS: Fifteen healthy laboratory beagles. METHODS: All dogs were video-recorded for 270 min after two intravenous recombinant IL-31 injections (1.75 µg/kg) and vehicle (phosphate-buffered saline, control) injections, respectively; interventions were randomised and performed with a 2 week wash-out period. Two blinded investigators reviewed the pruritic behaviours of all video recordings. RESULTS: Both canine IL-31 (IL-31_01, IL-31_02) injections significantly increased pruritic seconds and categorical minutes ('YES'/'NO' behaviour per discrete 1 min interval) in healthy dogs compared with both vehicle groups (Vehicle_01, Vehicle_02). The second intravenous canine IL-31 (IL-31_02) administered 14 days after the first IL-31 injection induced a significant increase in pruritic seconds (p = 0.021) and not pruritic categorical minutes (p = 0.231). An increase in pruritic seconds was observed in both IL-31 groups in the first 30 min post-administration, while there was no significant difference between IL-31 and vehicle groups. CONCLUSIONS AND CLINICAL RELEVANCE: In conclusion, intravenous IL-31 reproducibly induces itch responses in dogs. Future evaluations of the canine IL-31 pruritic model should assess total pruritic behaviours in seconds rather than using a biased 'YES/NO' behaviour per 1 min scoring system.

Effect of glycerin on immediate cutaneous reactions of histamine (positive) and phosphate-buffered saline (negative) controls in intradermal skin testing of healthy dogs: A randomised, blinded study.

BACKGROUND: Glycerinated allergen extracts contain 50% glycerin, an excellent preservative. While glycerin is a recognised irritant in humans, the utility of glycerinated extracts for intradermal testing has not been validated in dogs. HYPOTHESIS/OBJECTIVE: To determine and compare the effects of glycerin on immediate cutaneous reactions to intradermal injections of histamine and saline in healthy dogs. ANIMALS: Eight healthy laboratory beagles. MATERIALS AND METHODS: The study was designed as a randomised, blinded study. Intradermal injections of histamine (positive control) and saline (negative control) in aqueous and glycerinated (50%) forms were performed on the right thorax. Global wheal scores (GWS) at 20 min were evaluated by two independent investigators blinded to the interventions. RESULTS: There were no wheal and flare reactions observed after the intradermal injections of phenolated saline. By contrast, 50% glycerosaline injections induced erythema and induration in all dogs. Global wheal scores were significantly higher in aqueous histamine (Friedman test, p < 0.0001) and 50% glycerinated histamine (Friedman test, p = 0.0084) compared to phenolated saline controls. Interestingly, only aqueous histamine (Friedman test, p = 0.01) had significantly higher GWS than 50% glycerosaline injections, while no significant difference in GWS between 50% glycerinated histamine and 50% glycerosaline groups was observed (Friedman test, p = 0.59). CONCLUSION AND CLINICAL RELEVANCE: This study demonstrates that intradermal injection of 50% glycerosaline induces erythema and induration skin reactions in healthy dogs that can mimic positive reactions to allergenic extracts. Further dilutions of glycerinated positive and negative control solutions need to be optimised for intradermal testing in dogs.

Effect of diphenhydramine and cetirizine on immediate and late-phase cutaneous allergic reactions in healthy dogs: a randomized, double-blinded crossover study.

BACKGROUND: Currently, there is insufficient evidence to confirm oral diphenhydramine (DPH) efficacy to prevent mast cell degranulation and histamine release in dogs. HYPOTHESIS/OBJECTIVE: To determine and compare the effects oral of DPH and cetirizine on the immediate- and late-phase cutaneous allergic reactions in healthy dogs. ANIMALS: Twelve healthy laboratory beagle dogs. METHODS AND MATERIALS: The study was designed as a randomized, double-blinded crossover study in which each dog served as its own control; twice-daily oral DPH (2.2 mg/kg) or cetirizine (2 mg/kg) were given for six days with a two week washout period. Intradermal injections of histamine, compound 48/80 (positive control) and saline (negative control) were performed on the right thorax 10 days before drug administration (baseline), during oral antihistamine administration on Day 6 and 10 days after last medication dosage. Global wheal scores (GWS) at 20 min and late-phase reactions (LPR) at 6 h post-injection were evaluated by an investigator blinded to the drug and the interventions. RESULTS: Treatment with cetirizine significantly reduced histamine and compound 48/80 GWS and LPR compared to baseline; there was no significant difference for DPH. In all dogs, oral DPH and cetirizine reached plasma concentrations considered therapeutic in people. No adverse effect or behavioural changes were observed during the study. CONCLUSION AND CLINICAL SIGNIFICANCE: In conclusion, oral cetirizine was effective in preventing cutaneous allergic reactions without any obvious adverse effects in dogs. Oral DPH failed to show an inhibitory effect despite attaining plasma drug concentrations that are considered effective in people.

Characterization of a chloroquine-induced canine model of pruritus and skin inflammation.

BACKGROUND: Chloroquine (CQ) is a prototypical systemic and intradermal pruritogen for histamine-independent (nonhistaminergic) itch in mice and humans. The predictive validity of this model is poorly documented in dogs. HYPOTHESIS/OBJECTIVE: To determine pruritogenic and inflammatory effects of systemic and i.d. CQ injections in healthy dogs. ANIMALS: Ten healthy purpose-bred laboratory beagles. METHODS AND MATERIALS: All dogs were randomized to receive i.d. (200 and 400 µg/site), intravenous (2 mg/kg) and subcutaneous (3 mg/kg) CQ injections. Dogs were video-recorded for 30 min after i.d. injections and for 300 min after i.v. and s.c. injections. Buffered saline injections served as controls for each route. Global wheal scores were evaluated at 30 min post-i.d. injection by a blinded investigator. RESULTS: All dogs showed wheal and erythema at the CQ i.d. injection sites; global wheal scores of each CQ concentration were significantly increased compared to placebo (P ≤ 0.05). Blinded evaluation revealed no significant increase in generalized pruritic behaviour (pruritic seconds) after i.v. or s.c. administration of CQ. Intradermal injections induced mild localized acute pruritic behaviours at the site of injections at 200 µg (P = 0.06) and 400 µg (P = 0.27) CQ in dogs. CONCLUSION AND CLINICAL SIGNIFICANCE: To the best of the authors' knowledge, this is the first report which shows that i.d. CQ injections may induce acute inflammation in healthy dogs. By contrast to the systemic CQ-induced pruritus reported previously in healthy mice and dogs, no significant pruritic behaviours were observed after CQ injection, regardless of the route of administration.

Dose-dependent pruritogenic and inflammatory effects of intradermal injections of histamine, compound 48/80 and anti-canine IgE in healthy dogs.

BACKGROUND: Scratching behaviours associated with intradermal (i.d.) injection of pruritogens such as histamine and compound 48/80 into the skin of mice and humans is the commonly used model to advance itch research and drug development. The predictive validity of this model is poorly documented in dogs. OBJECTIVES: To evaluate the dose-dependent effects of pruritogenic substances, each with a different mechanism of action, in healthy dogs. ANIMALS: Ten healthy laboratory beagles. METHODS AND MATERIALS: All dogs were video-recorded for 30 min post-injection (mpi) of i.d. goat anti-canine IgE (4 and 25 µg/site), histamine and compound 48/80 (50, 100, 200, 400 µg/site); two buffered saline injections served as controls. Two blinded investigators reviewed the pruritic behaviours of all video recordings. Global wheal scores were evaluated at 30 min by a blinded investigator. RESULTS: All dogs showed wheal and erythema at the pruritogen injection site; global wheal scores at 30 min of each substance significantly increased at all concentrations compared to control (P ≤ 0.05). A blinded evaluation revealed that all pruritogens induced mild acute pruritic behaviours at the site of injection. There was no injection site pain seen in any dog. Compared to controls, injections of pruritogens did not significantly affect the pruritic seconds or occurrence of pruritic episodes for any of the substances. CONCLUSIONS AND CLINICAL SIGNIFICANCE: These preliminary results suggest that i.d. injections of the studied pruritogens can induce cutaneous wheal and flare response in healthy dogs; but inconsistencies occur in the induction of itch, even with the different concentrations of pruritogens.

The anti-inflammatory effect of topical tofacitinib on immediate and late-phase cutaneous allergic reactions in dogs: a placebo-controlled pilot study.

BACKGROUND: Topical Janus kinase (JAK) inhibition is a promising therapeutic target for several inflammatory skin diseases of humans. OBJECTIVES: To evaluate the anti-inflammatory effect of tofacitinib, a JAK 1/3 inhibitor, on immediate and late-phase skin reactions in dogs. ANIMALS: Five healthy laboratory beagle dogs. METHODS: Topical tofacitinib (total daily dosage: 0.5 mg/cm2 ) or its gel vehicle were applied on either the left or right lateral thorax of each dog for eight days. Three days before application and after eight days of topical treatment, intradermal injections of histamine and anticanine-IgE antibodies were performed on both sides; they were evaluated by an investigator blinded to the interventions. RESULTS: The tofacitinib gel was well-tolerated; one dog developed mild erythema at Day 5 that resolved by the next application. Treatment with tofacitinib reduced histamine and anticanine-IgE global wheal scores (one-way ANOVA, P ≤ 0.005 for both) compared to baseline; there was no significant difference for the vehicle placebo (histamine; P = 0.163; IgE, P = 0.223). Late-phase reactions (LPRs) were markedly, but not significantly reduced after tofacitinib treatment (P = 0.071). A blinded histological evaluation of 6 h-anti-IgE-associated LPRs revealed a significant reduction in the total leucocyte superficial dermal cellularity (P = 0.022), as well as eosinophil (P = 0.022) and mast cell (P = 0.022) counts at tofacitinib-treated sides compared with pretreatment values. Post-treatment complete blood counts and serum chemistry profiles did not show relevant tofacitinib-induced changes. CONCLUSIONS: Our observations suggest that topical tofacitinib exerts an inhibitory effect on activated canine skin-emigrating immune cells; this drug should be investigated further as a topical immunosuppressive drug in dogs.