title: Plasmodium falciparum: the adherence of erythrocytes infected with human malaria can be mimicked using pfalhesin-coated microspheres.
abstract: Infection of human erythrocytes with the malaria parasite, Plasmodium falciparum, results in the exposure of amino acid residues 542-555 of the anion-exchange protein, band 3, in a conformation that enables the cell to adhere to C32 amelanotic melanoma cells. Attempts to isolate this adhesive form from infected cells by immunoaffinity were unsuccessful, and so other approaches were utilized. Chinese hamster ovary (CHO) cells transfected with cDNA encoding the first 578 amino acid residues of human band 3 protein transiently expressed the protein efficiently. A murine monoclonal antibody (MAb) that specifically recognizes the adhesin exposed on the surface of erythrocytes bearing mature stages of P. falciparum immunostained some transfected cells, confirming that the first 578 amino residues are sufficient for the adhesive conformation. As a more efficient alternative to transgenic expression of the adhesin, microspheres with covalently bound peptides fashioned on band 3 sequences previously found to be adherent (residues 546-553 and 820-829 and called pfalhesin) were produced. The pfalhesin-coated microspheres specifically bound to C32 amelanotic melanoma cells, whereas microspheres coupled with a scrambled version of residues 546-553 had little binding capacity for melanoma cells. These results demonstrate that the previously identified band 3-related peptides that inhibit cytoadherence interact directly with target cells and suggest that microspheres with covalently coupled peptides might constitute novel 'artificial' P. falciparum-infected erythrocytes for use in in vitro and in vivo studies.
authors.affiliation: Department of Biology, University of California, Riverside 92521, USA.
authors.forename: N
authors.initials: N
authors.lastname: Guthrie
authors.forename: D M
authors.initials: DM
authors.lastname: Bird
authors.forename: I
authors.initials: I
authors.lastname: Crandall
authors.forename: I W
authors.initials: IW
authors.lastname: Sherman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D016466:CHO Cells / Q000469:parasitology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D006224:Cricetinae; D010450:Endopeptidases; D004912:Erythrocytes / Q000166:cytology / Q000469:parasitology*; D006801:Humans; D016778:Malaria, Falciparum / Q000097:blood; D008545:Melanoma; D008863:Microspheres; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D010963:Plasmodium falciparum / Q000166:cytology*; D011487:Protein Conformation; D014407:Tumor Cells, Cultured / Q000378:metabolism
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; C086125:pfalhesin; D010450:Endopeptidases