CD47 regulates the phagocytic clearance and replication of the Plasmodium yoelii malaria parasite

Several Plasmodium species exhibit a strong age-based preference for the red blood cells (RBC) they infect, which in turn is a major determinant of disease severity and pathogenesis. The molecular basis underlying this age constraint on the use of RBC and its influence on parasite burden is poorly understood. CD47 is a marker of self on most cells, including RBC, which, in conjunction with signal regulatory protein alpha (expressed on macrophages), prevents the clearance of cells by the immune system. In this report, we have investigated the role of CD47 on the growth and survival of nonlethal Plasmodium yoelii 17XNL (PyNL) malaria in C57BL/6 mice. By using a quantitative biotin-labeling procedure and a GFP-expressing parasite, we demonstrate that PyNL parasites preferentially infect high levels of CD47 (CD47^hi)-expressing young RBC. Importantly, C57BL/6 CD47^−/− mice were highly resistant to PyNL infection and developed a 9.3-fold lower peak parasitemia than their wild-type (WT) counterparts. The enhanced resistance to malaria observed in CD47^−/− mice was associated with a higher percentage of splenic F4/80⁺ cells, and these cells had a higher percentage of phagocytized parasitized RBC than infected WT mice during the acute phase of infection, when parasitemia was rapidly rising. Furthermore, injection of CD47-neutralizing antibody caused a significant reduction in parasite burden in WT C57BL/6 mice. Together, these results strongly suggest that CD47^hi young RBC may provide a shield to the malaria parasite from clearance by the phagocytic cells, which may be an immune escape mechanism used by Plasmodium parasites that preferentially infect young RBC.Malaria, caused by Plasmodium parasites, remains a major cause of mortality and morbidity in the developing world. Among the four principal human Plasmodium species, Plasmodium falciparum is the most virulent, being responsible for more than 90% of malaria-associated deaths. Likewise, Plasmodium species that infect rodents and nonhuman primates also differ widely in their fulminant nature and in the mortality they cause (1–3). How different Plasmodium species have evolved to exhibit this wide array of virulence and disease severity remains one of the major unsolved questions in malaria biology and pathogenesis.One important factor that is associated with Plasmodium parasite burden and disease severity is the age constraint of the host red blood cells (RBC) they infect. The age-based preference for restricted invasion of RBC by the Plasmodium parasite is characterized as young RBC (reticulocyte), aged RBC (mature), or both young and aged RBC. Plasmodium species that preferentially infect and grow inside young RBC generally cause a low-grade, self-resolving infection that is rarely fatal (e.g., Plasmodium vivax and Plasmodium ovale), whereas those that infect both young and aged RBC cause more fulminant infection that can be fatal in the absence of immunity (e.g., P. falciparum) (1, 4–6). Thus, along with host genetic background and immune response, restriction for age-specific RBC invasion is a major determinant of the severity and outcome of malaria infection.Malaria parasites have evolved to use redundant receptors and pathways to invade the RBC. For example, sialic acid (7) and Duffy antigen (8) are the major RBC receptors for invasion of P. falciparum and P. vivax, respectively, although other receptors and invasion pathways are known to exist (9, 10). Although a redundancy in RBC receptor use would ensure successful invasion by mitigating the effects of polymorphism and immune targeting, the reasons behind the RBC age-based preference for invasion are not fully clear and remain a subject of debate.Survival of normal cells through the course of their life cycle is essential to maintain homeostasis, and aberrant cells (e.g., senescent or foreign antigen-expressing cells) are eliminated through a sophisticated programmed cell removal system that relies on the recognition of self and nonself determinants (11). CD47, a cell surface molecule in the Ig superfamily, is ubiquitously expressed on many cell types, including RBC, and is a marker of self to avoid early clearance by phagocytic cells through ligation of signal regulatory protein alpha (12). In contrast, altered expression or conformational changes in CD47 may lead to a molecular switch that triggers a phagocytic signal to remove aged or damaged cells (11). Recent studies have shown that the level of CD47 expression is higher in progenitor cells and declines as they undergo maturation and are subsequently aged (13). This age-dependent difference in CD47 expression shields young cells but allows clearance of aging and damaged cells from the system.CD47 is overexpressed in cancer cells (11, 14, 15), and the CD47–signal regulatory protein alpha interaction is considered a major pathway of immune evasion by tumor cells (15). Administration of anti-CD47 antibodies enabled the phagocytosis of tumor cells in vitro, reduced their growth, and prevented the metastasis of human patient tumor cells (14). In this article, using the murine Plasmodium yoelii nonlethal model, we provide quantitative evidence for age of RBC as the basis for the survival and growth of malaria parasites and provide supporting data that suggest that P. yoelii nonlethal parasites prefer to grow inside younger RBC, which allows them to evade immune clearance by phagocytic cells through a CD47-mediated process, and that CD47 modulates the clearance of malaria infection. To our knowledge, this is the first report that provides a molecular basis for the age-dependent preference for infection of RBC by a Plasmodium parasite and sheds light on its implications for the severity of malaria infection in a host.