Allopurinol therapy provides long term clinical improvement, but additional immunotherapy is required for sustained parasite clearance, in L. infantum-infected dogs

Leopoldo F M Nascimento; Dayane Francisca Higino Miranda; Luana D Moura; Flaviane A Pinho; Guilherme Loureiro Werneck; Ricardo Khouri; Steven G Reed; Malcolm S Duthie; Aldina Barral; Manoel Barral-Netto; Maria S P Cruz

doi:10.1016/j.jvacx.2019.100048

Allopurinol therapy provides long term clinical improvement, but additional immunotherapy is required for sustained parasite clearance, in L. infantum-infected dogs

Vaccine X. 2019 Nov 20:4:100048. doi: 10.1016/j.jvacx.2019.100048. eCollection 2020 Apr 9.

Authors

Leopoldo F M Nascimento¹, Dayane Francisca Higino Miranda¹, Luana D Moura¹, Flaviane A Pinho², Guilherme Loureiro Werneck³, Ricardo Khouri^{4

2}, Steven G Reed^{5

6}, Malcolm S Duthie^{5

6}, Aldina Barral^{4

2

7}, Manoel Barral-Netto^{4

2

7}, Maria S P Cruz¹

Affiliations

¹ Universidade Federal do Piauí, Departamento de Morfofisiologia Veterinária, Teresina, PI, Brazil.
² Universidade Federal da Bahia, Faculdade de Medicina, Salvador, BA, Brazil.
³ Universidade Federal do Rio de Janeiro, Instituto de Estudos em Saúde Coletiva, Rio de Janeiro, RJ, Brazil.
⁴ Fundação Oswaldo Cruz- Fiocruz, Instituto Gonçalo Moniz, Salvador, BA, Brazil.
⁵ Infectious Diseases Research Institute, Seattle, WA 98102, USA.
⁶ HDT Biotech Corporation, Seattle, WA 98102, USA.
⁷ Instituto Nacional de Ciência e Tecnologia, Instituto de Investigação em Imunologia, São Paulo, SP, Brazil.

Abstract

There is little evidence that current control strategies for canine leishmaniosis (CanL), the veterinary disease caused by L. infantum infection, are having a positive impact. This is of critical importance because dogs are a primary reservoir for L. infantum and a significant source of parasite transmission to humans. Drugs intended primarily for human use are prohibited for the treatment of CanL because of concerns over the propagation of resistant parasites. Although allopurinol effectively decreases parasite burden in CanL the treatment needs to be maintained for life. We hypothesized that during the allopurinol-induced parasite reduction dogs may become capable of developing a more robust immune response that may permit more effective control of parasites. To test this, we investigated the clinical and parasitological impact of short-term treatment with allopurinol, either alone or in combination with a defined subunit vaccine, on dogs naturally infected with L. infantum. A total of 28 dogs were distributed as follows: untreated; oral allopurinol alone (20 mg/kg, once each day for 90 days); or allopurinol with immunization with the Leish-F2 antigen formulated with the Toll-like receptor (TLR) 4 agonist Second generation Lipid Adjuvant (SLA) in stable emulsion (SE; SLA-SE). Dogs that did not receive treatment had a progressive decline in their clinical condition and an increase in their infection levels, while treatment with allopurinol alone alleviated the clinical symptoms of CanL but did not generate sustained reduction in parasites. Concomitant immunization with Leish-F2 + SLA-SE, however, improved clinical condition while also providing long-term clearance of L. infantum from lymphoid tissues and systemic organs. These results have important implications for both the management of CanL and for limiting L. infantum transmission to humans.

Keywords: CanL, canine leishmaniosis; Canine visceral leishmaniasis; Clinical signs; Drug; GLA, glycopyranosyl lipid; IFN, interferon; IL, interleukin; MPL, monophosphoryl lipid A; Parasite; SE, stable emulsion; SLA, Second generation Lipid Adjuvant; TLR, Toll-like receptor; Th1, T helper 1-like cells; VL, visceral leishmaniasis; Vaccine.

Grants and funding

R01 AI025038/AI/NIAID NIH HHS/United States