Last Updated: 29/01/2025
Developmental research aimed at establishing a practical in vitro culture system for Plasmodium vivax
Objectives
*Original title and text were machine translated from Japanese.
This research project aims to create a P. vivax parasite that can invade and grow in human mature red blood cells by introducing a ligand that recognizes human mature red blood cells into the parasite. It is believed that by using this parasite, a practical culture system can be established for the parasite.
Plasmodium vivax invades and grows in human immature red blood cells, but since it is difficult to routinely obtain large amounts of human immature red blood cells, there is no practical culture system for P. vivax. Further, the plan is to evaluate the ligand expression plasmid produced using P. cynomolgi and to examine the phenotype in P. cynomolgi.
In the first year, an EBA175 expression plasmid was introduced into P. cynomolgi, but no drug-resistant parasites appeared even after three months. Therefore, gene introduction experiments were repeated using multiple protocols, including episomal gene introduction and CRISPR/Cas9 gene editing. However, drug-resistant strains could not be obtained. Since gene transfer into P. knowlesi was performed without any problems, this seems to be a problem specific to P. cynomolgi or the P. cynomolgi strain that was used. In addition, the P. cynomolgi that was used grows slower than the P. knowlesi and Plasmodium falciparum that are maintained in vitro, so it took time to evaluate whether gene transfer was successful, and recombinant P. cynomolgi parasites could not be obtained during this year. On the other hand, attempt was made to establish in vitro culture strains of P. knowlesi from the blood of several human patients infected with P. knowlesi using the culture medium optimized for P. cynomolgi established in the first year, and the team succeeded in establishing human strains.
Jun 2022 — Mar 2025
$44,422
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