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During the clinically silent liver stage of a  Plasmodium  infection the parasite replicates from a single sporozoite into thousands of merozoites. Infection of humans and rodents with large numbers of sporozoites that arrest their development within the liver can cause sterile protection from subsequent infections. Disruption of genes essential for liver stage development of rodent malaria parasites has yielded a number of attenuated parasite strains. A key question to this end is how increased attenuation relates to vaccine efficacy. Here, we generated rodent malaria parasite lines that arrest during liver stage development and probed the impact of multiple gene deletions on attenuation and protective efficacy. In contrast to  P. berghei  strain ANKA  LISP2 (–) or  uis3 (–) single knockout parasites, which occasionally caused breakthrough infections, the double mutant lacking both genes was completely attenuated even when high numbers of sporozoites were administered. However, different vaccination protocols showed that  LISP2 (–) parasites protected better than  uis3 (–) and double mutants. Hence, deletion of several genes can yield increased safety but might come at the cost of protective efficacy.

Protective efficacy and safety of liver stage attenuated malaria parasites