Leishmania HASP and SHERP Genes Are Required for In Vivo Differentiation, Parasite Transmission and Virulence Attenuation in the Host

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Abstract
Differentiation of extracellular Leishmania promastigotes within their sand fly vector, termed metacyclogenesis, is considered to be essential for parasites to regain mammalian host infectivity. Metacyclogenesis is accompanied by changes in the local parasite environment, including secretion of complex glycoconjugates within the promastigote secretory gel and colonization and degradation of the sand fly stomodeal valve. Deletion of the stage-regulated HASP and SHERP genes on chromosome 23 of Leishmania major is known to stall metacyclogenesis in the sand fly but not in in vitro culture. Here, parasite mutants deficient in specific genes within the HASP/SHERP chromosomal region have been used to investigate their role in metacyclogenesis, parasite transmission and establishment of infection. Metacyclogenesis was stalled in HASP/SHERP mutants in vivo and, although still capable of osmotaxis, these mutants failed to secrete promastigote secretory gel, correlating with a lack of parasite accumulation in the thoracic midgut and failure to colonise the stomodeal valve. These defects prevented parasite transmission to a new mammalian host. Sand fly midgut homogenates modulated parasite behaviour in vitro, suggesting a role for molecular interactions between parasite and vector in Leishmania development within the sand fly. For the first time, stage-regulated expression of the small HASPA proteins in Leishmania (Leishmania) has been demonstrated: HASPA2 is expressed only in extracellular promastigotes and HASPA1 only in intracellular amastigotes. Despite its lack of expression in amastigotes, replacement of HASPA2 into the null locus background delays onset of pathology in BALB/c mice. This HASPA2-dependent effect is reversed by HASPA1 gene addition, suggesting that the HASPAs may have a role in host immunomodulation. Millions of people around the world are at risk of infection with single-celled Leishmania parasites that cause a wide range of infectious diseases of the immune system, the leishmaniases. There is no effective vaccine for these infections while available drugs have toxic side-effects and resistance is an increasing problem. Human infection occurs through the bite of infected blood-feeding sand flies. Leishmania parasites live in the sand fly gut, where they complete a complex series of developmental changes to become infective to mammals. The parasites also modify the insect gut to promote their own transmission. Little is known about the molecular regulation of these processes. Recently, we showed that deletion of a small group of related genes from the parasite prevented completion of its development in the sand fly, suggesting a role for these sequences in transmission to the host. This study clarifies the expression pattern of these genes during parasite development and shows that the observed stalling of development is accompanied by changes in parasite-sand fly gut interactions and a loss of parasite transmission. These target genes also influence disease development in the mammalian host, confirming critical roles for their encoded proteins throughout the parasite life cycle.
Funding Information
  • Wellcome Trust (077503)
  • Biotechnology and Biological Sciences Research Council
  • COST-CZ (LD14076)
  • COST-CZ (LD14076)
  • COST-CZ (LD14076)
  • EurNegVec COSTAction (TD1303)
  • EurNegVec COSTAction (TD1303)
  • EurNegVec COSTAction (TD1303)
  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases
  • National Institute of Allergy and Infectious Diseases

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