PfEMP1 PfEMP1

Immunofluorescence assay with live infected or formalin-fixed PaloAlto VarO erythrocytes. Live Palo Alto VarO mature infected erythrocytes (pigmented trophozoites and schizonts) (A-D) or formalin-fixed cultures (E) were incubated with mouse antisera against PfEMP1-VarO-derived recombinant domains at 1/200 dilution, and bound antibodies visualised using Alexa Fluor 488-labelled goat anti-mouse IgG (Invitrogen) at 1/1,000 dilution and nuclei were labelled with Hoechst 33342 dye at 1/1,000. Typical punctate surface staining of the membrane of the infected erythrocyte (green) was observed with antisera raised to bDBL1, eDBL1, pCIDR, eDBL2, bDBL2, eDBL4 and eHead (representative examples are shown here in (B-D) as indicated. Antisera raised to eDBL0, which failed to react with the erythrocyte surface, reacted with a perinuclear image on formalin-fixed Palo Alto VarO parasites (E). Preimmune sera to formalin-fixed parasites were negative (not shown). The parasite nuclei are stained with DAPI (blue). Slides were viewed with a 100x objective using a Leica CTR5000 fluorescent microscope .

Guillotte M, Juillerat A, Igonet S, Hessel A, Petres S, Crublet E, Le Scanf C, Lewit-Bentley A, Bentley GA, Vigan-Womas I, Mercereau-Puijalon O. Immunogenicity of the Plasmodium falciparum PfEMP1-VarO Adhesin: Induction of Surface-Reactive and Rosette-Disrupting Antibodies to VarO Infected Erythrocytes. PLoS One. 2015 10(7):e0134292.

img