Long-term live imaging reveals cytosolic immune responses of host hepatocytes against Plasmodium infection and parasite escape mechanisms

• Published in: Autophagy 2015, Vol.11 (9), p.1561-1579
• Main Authors: Prado, Monica, Eickel, Nina, De Niz, Mariana, Heitmann, Anna, Agop-Nersesian, Carolina, Wacker, Rahel, Schmuckli-Maurer, Jacqueline, Caldelari, Reto, Janse, Chris J, Khan, Shahid M, May, Jürgen, Meyer, Christian G, Heussler, Volker T
• Format: Article
• Language: English
• Subjects: Adaptor Proteins, Signal Transducing - metabolism; Animals; Autophagy; Basic Research Paper; Biomarkers - metabolism; Cytosol - immunology; cytosolic immune response; Galectins - metabolism; Heat-Shock Proteins - metabolism; Hep G2 Cells; Hepatocytes - immunology; Hepatocytes - parasitology; Hepatocytes - ultrastructure; Humans; Imaging, Three-Dimensional; Immune Evasion; Immunity; Intracellular Membranes - metabolism; Intracellular Membranes - ultrastructure; LC3; Life Cycle Stages; Liver - parasitology; long-term live imaging; Lysosomes - metabolism; Lysosomes - ultrastructure; malaria; Malaria - immunology; Malaria - parasitology; Mice; Microtubule-Associated Proteins - metabolism; Parasites - growth & development; Parasites - immunology; Parasites - pathogenicity; Parasites - ultrastructure; Plasmodium berghei - growth & development; Plasmodium berghei - pathogenicity; Plasmodium berghei - ultrastructure; Plasmodium liver-stage; selective autophagy; Sequestosome-1 Protein; Sporozoites - physiology; Sporozoites - ultrastructure; Survival Analysis; Time Factors; ubiquitin; Ubiquitin - metabolism; Ubiquitination; Vacuoles - metabolism; Vacuoles - ultrastructure; Virulence
• ISSN: 1554-8627; 1554-8635
• DOI: 10.1080/15548627.2015.1067361

Plasmodium parasites are transmitted by Anopheles mosquitoes to the mammalian host and actively infect hepatocytes after passive transport in the bloodstream to the liver. In their target host hepatocyte, parasites reside within a parasitophorous vacuole (PV). In the present study it was shown that the parasitophorous vacuole membrane (PVM) can be targeted by autophagy marker proteins LC3, ubiquitin, and SQSTM1/p62 as well as by lysosomes in a process resembling selective autophagy. The dynamics of autophagy marker proteins in individual Plasmodium berghei-infected hepatocytes were followed by live imaging throughout the entire development of the parasite in the liver. Although the host cell very efficiently recognized the invading parasite in its vacuole, the majority of parasites survived this initial attack. Successful parasite development correlated with the gradual loss of all analyzed autophagy marker proteins and associated lysosomes from the PVM. However, other autophagic events like nonselective canonical autophagy in the host cell continued. This was indicated as LC3, although not labeling the PVM anymore, still localized to autophagosomes in the infected host cell. It appears that growing parasites even benefit from this form of nonselective host cell autophagy as an additional source of nutrients, as in host cells deficient for autophagy, parasite growth was retarded and could partly be rescued by the supply of additional amino acid in the medium. Importantly, mouse infections with P. berghei sporozoites confirmed LC3 dynamics, the positive effect of autophagy activation on parasite growth, and negative effects upon autophagy inhibition.