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

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.

Highlights

To evaluate if chemo- and or chemoimmunotherapeutic intervention could interrupt L. infantum infection and canine leishmaniosis (CanL) development, we enrolled dogs that were confirmed to be infected with L. infantum
We observed that allopurinol, either alone or in conjunction with defined subunit vaccine LeishF2 + second generation lipid adjuvant (SLA)-stable emulsion (SE), provided long-term clinical improvement of the infected dogs
When we evaluated the infectious status, our data revealed that treatment with allopurinol alone initially reduced parasite burden in the bone marrow, one year later numbers had rebounded to pre-treatment levels and infection was disseminated throughout the body

Summary

Introduction

⇑ Corresponding author.Visceral leishmaniasis (VL), the clinical consequence of infection with Leishmnaia infantum, is a serious public health problem in several countries. Dogs are incriminated as the main domestic reservoirs of L. infantum and are responsible for sustained transmission to humans [1]. Several measures have been recommended to combat CanL with a goal of limiting parasite transmission and disease in humans. In Brazil, due to the perceived risk of the potential emergence of drug resistant parasites, treatment of CanL is regulated by Interministerial Order No 1426 that prohibits its treatment with drugs intended primarily for human use [2]. Vaccines can be used more broadly than drugs, and could be used to protect both animals and humans. The vaccines used for Leishmania, have, demonstrated varying efficacies when evaluated by prophylactic or therapeutic approaches [3]

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Results

Conclusion