Malaria parasites: Oral and Nasal inoculation of mice with iRBC’s & development of P. falciparum mutants with the CRISPR/cas9 system

Electronic versions

Dogfennau

  • Catia Antunes

Abstract

Malaria is a parasitic disease caused by parasites of the Plasmodium genus and in 2015 around 214,000,000 new cases of malaria infection occurred resulting in 438,000 deaths worldwide. Currently, there is no available vaccine against malaria and despite efforts, mainly channelled into subunit vaccines, immunogenicity is often low. In this work, whole live blood stage Plasmodium was used in oral and nasal inoculation of mice to test for infectivity and immunogenicity. Oral inoculation resulted in infection of some mice while none of those inoculated nasally became infected. Both oral and nasal inoculation, despite not providing protection, allowed for extended survival of inoculated individuals. Given that survival was increased by inoculating animals solely with live parasites, the results detailed in this work show that this approach holds great applicational potential, as coupling oral and nasal inoculation with live parasites and a powerful adjuvant may be the key to unlock full protection. In connection with inoculation of live parasites, this work also aimed at producing a Plasmodium falciparum mutant expressing Salmonella’s FimH+ glycoprotein as to allow its detection by M cells in the gut. This was to be achieved by a modified version of the CRISPR/Cas9 system which would bear a double guide RNA instead of a single. However, this was not possible as the resulting CRISPR plasmid contained an extremely high AT-content becoming toxic to E. coli. In addition to these two main goals, a few molecular techniques crucial for malaria research were improved and a detection method of avian malaria was developed.

Details

Iaith wreiddiolSaesneg
Sefydliad dyfarnu
Goruchwylydd / Goruchwylwyr / Cynghorydd
Dyddiad dyfarnuIon 2017