Plasmodium P36 determines host cell receptor usage during sporozoite invasion
- Sorbonne Universités, UPMC Univ Paris 06, INSERM, CNRS, Centre d’Immunologie et des Maladies Infectieuses, U1135, ERL8255, France
- Mahidol University, Thailand
- University of Oxford, United Kingdom
- INSERM, U1110, Institut de Recherche sur les Maladies Virales et Hépatiques, France
- Université de Strasbourg, France
- Sorbonne Universités, UPMC Univ Paris 06, INSERM, Institute of Cardiometabolism and Nutrition, UMR_S 1166, France
- INSERM, U935, France
- Université Paris Sud, Institut André Lwoff, France
- Assistance Publique Hôpitaux de Paris, Centre Hospitalo-Universitaire Pitié-Salpêtrière, France
- Institut Hospitalo-Universitaire, Pôle Hépato-digestif, Hopitaux Universitaires de Strasbourg, France
Figures
Figure 1
Anti-SR-BI antibodies inhibit P. vivax but not P. falciparum sporozoite infection.
(A) Primary human hepatocyte cultures were incubated with P. vivax sporozoites in the presence of anti-CD81 and/or anti-SR-BI mAbs at 20 μg/ml, and the number of EEF-infected cells was determined 5 …
Figure 2 with 3 supplements
Infection of human HepG2 cells by P. berghei sporozoites depends on SR-BI.
(A) HepG2 cells were incubated with P. berghei sporozoites for 3 hr in the absence (Control) or presence of increasing concentrations of rabbit polyclonal SR-BI antisera. Infected cultures were …
Figure 2—figure supplement 1
CD81 and SR-BI surface expression in HepG2 and HepG2/CD81 cells.
HepG2 (A) and HepG2/CD81 cells (B) were stained with anti-CD81 (red histograms) or anti-SR-BI (green histograms) antibodies and analyzed by FACS.
Figure 2—figure supplement 2
Anti-SR-BI antibodies neutralize P. berghei infection of HepG2 cells.
Effect of anti-SR-BI rat polyclonal antibodies (pAb) and mouse mAb on P. berghei EEF numbers in HepG2 cells.
Figure 2—figure supplement 3
Effect of SR-BI silencing on sporozoite cell traversal and invasion.
HepG2 cells transfected with siRNA oligonucleotides targeting SR-BI (siSR-BI) or a control siRNA (siCont) were incubated for 3 hr with PbGFP sporozoites and rhodamine-labeled dextran, and the …
Figure 3 with 1 supplement
CD81 and SR-BI define alternative entry routes for P. berghei sporozoites.
(A) HepG2/CD81 cells were incubated with P. berghei sporozoites in the presence or absence of anti-human CD81 and/or SR-BI mAbs, and the number of EEFs-infected cells was determined by fluorescence …
Figure 3—figure supplement 1
Effect of anti-CD81 and anti-SR-BI antibodies on P. berghei sporozoite cell traversal and intracellular development.
(A) HepG2/CD81 cells were incubated for 3 hr with PbGFP sporozoites and rhodamine-labeled dextran, in the presence or absence of anti-CD81 and/or SR-BI antibodies. The percentage of traversed …
Figure 4
Infection of HepG2/CD81 cells by P. yoelii sporozoites depends on CD81 but not SR-BI.
(A) HepG2/CD81 cells were incubated with P. yoelii sporozoites in the presence of anti-CD81 mAb (20 μg/ml) or anti-SR-BI polyclonal rabbit serum (diluted 1/100), and the number of EEF-infected cells …
Figure 5 with 4 supplements
The 6-cys proteins P52 and P36 are required for productive host cell invasion.
(A–B) HepG2 (A) or Hepa1-6 cells (B) were incubated with PbGFP (blue lines) or PbΔp52/p36 sporozoites (red lines) for 15–120 min, in the presence (dotted lines) or absence (solid lines) of …
Figure 5—figure supplement 1
Targeted gene deletion of p52 and p36 in P. berghei.
(A) Replacement strategy to generate PbΔp52p36 parasites. The wild-type (WT) genomic locus of P. berghei p52/p36 was targeted with a GOMO-GFP replacement plasmid containing a 5’ and a 3’ homologous …
Figure 5—figure supplement 2
Targeted gene deletion of p52 and p36 in P. yoelii.
(A) Replacement strategy to generate PyΔp52p36 parasites. The wild-type (WT) genomic locus of P. yoelii p52/p36 in the PyGFP parental line was targeted with a GOMO replacement plasmid containing a …
Figure 5—figure supplement 3
Targeted gene deletion of p36 in P. berghei.
(A) Replacement strategy to generate PbΔp36 parasites. The wild-type (WT) genomic locus of P. berghei p36 was targeted with a GOMO-GFP replacement plasmid containing a 5’ and a 3’ homologous …
Figure 5—figure supplement 4
P36 is required for P. berghei sporozoite entry via both SR-BI- and CD81-dependent routes.
HepG2 and HepG2/CD81 cells were incubated with PbGFP or PbΔp36 sporozoites in the presence or absence of anti-CD81 and anti-SR-BI antibodies, and the percentage of infected (GFP-positive) cells was …
Figure 6 with 1 supplement
P36 mediates CD81-independent entry in P. berghei sporozoites.
(A) HepG2 (blue histograms) or HepG2/CD81 cells (red histograms) were incubated with sporozoites from PbΔp52/p36 parasites genetically complemented with P. berghei and/or P. yoelii P52 and P36, and …
Figure 6—figure supplement 1
Genetic complementation with p52 and p36 from P. falciparum or P. vivax does not restore sporozoite infectivity in PbΔp52p36 parasites.
(A) HepG2 and HepG2/CD81 cells were incubated with PbGFP, PbΔp52p36 and PbΔp52p36 complemented with p52 and p36 from P. falciparum or P. vivax. The percentage of infected (GFP-positive) cells 24 hr …
Figure 7
Transgenic P. yoelii sporozoites expressing PbP36 can infect CD81-null cells via SR-BI.
(A) HepG2 (blue) and HepG2/CD81 cells (red) were incubated with genetically complemented PyΔp52/p36 sporozoites, and fixed 24 hr post-infection. The number of UIS4-positive vacuoles was then …
Figure 8 with 1 supplement
Model of host cell entry pathways for Plasmodium sporozoites.
(A) Host cell membrane proteins CD81 and SR-BI define two independent entry routes for Plasmodium sporozoites. P. falciparum and P. yoelii sporozoites require CD81 for infection, whereas P. vivax …
Figure 8—figure supplement 1
P36 protein sequence analysis.
(A) Alignment of P. berghei and P. yoelii P36 protein sequences. Identical, similar and different amino acids are shaded in black, grey and red, respectively. The tandem 6-cys domains D1 and D2 are …
