title: Antibodies from malaria-exposed pregnant women recognize trypsin resistant epitopes on the surface of Plasmodium falciparum-infected erythrocytes selected for adhesion to chondroitin sulphate A.
abstract: BACKGROUND
  The ability of Plasmodium falciparum-infected erythrocytes to adhere to the microvasculature endothelium is thought to play a causal role in malaria pathogenesis. Cytoadhesion to endothelial receptors is generally found to be highly sensitive to trypsinization of the infected erythrocyte surface. However, several studies have found that parasite adhesion to placental receptors can be markedly less sensitive to trypsin. This study investigates whether chondroitin sulphate A (CSA) binding parasites express trypsin-resistant variant surface antigens (VSA) that bind female-specific antibodies induced as a result of pregnancy associated malaria (PAM).
  
  
  METHODS
  Fluorescence activated cell sorting (FACS) was used to measure the levels of adult Scottish and Ghanaian male, and Ghanaian pregnant female plasma immunoglobulin G (IgG) that bind to the surface of infected erythrocytes. P. falciparum clone FCR3 cultures were used to assay surface IgG binding before and after selection of the parasite for adhesion to CSA. The effect of proteolytic digestion of parasite erythrocyte surface antigens on surface IgG binding and adhesion to CSA and hyaluronic acid (HA) was also studied.
  
  
  RESULTS
  P. falciparum infected erythrocytes selected for adhesion to CSA were found to express trypsin-resistant VSA that are the target of naturally acquired antibodies from pregnant women living in a malaria endemic region of Ghana. However in vitro adhesion to CSA and HA was relatively trypsin sensitive. An improved labelling technique for the detection of VSA expressed by CSA binding isolates has also been described.
  
  
  CONCLUSIONS
  The VSA expressed by CSA binding P. falciparum isolates are currently considered potential targets for a vaccine against PAM. This study identifies discordance between the trypsin sensitivity of CSA binding and surface recognition of CSA selected parasites by serum IgG from malaria exposed pregnant women. Thus, the complete molecular definition of an antigenic P. falciparum erythrocyte surface protein that can be used as a malaria in pregnancy vaccine has not yet been achieved.
authors.affiliation: Institute of Cell, Animal and Population Biology, University of Edinburgh, King's Buildings, West Mains Road, Edinburgh, EH9 3JT, Scotland, UK. L.sharling@sms.ed.ac.uk
authors.forename: Lisa
authors.initials: L
authors.lastname: Sharling
authors.forename: Anders
authors.initials: A
authors.lastname: Enevold
authors.forename: Kordai M P
authors.initials: KM
authors.lastname: Sowa
authors.forename: Trine
authors.initials: T
authors.lastname: Staalsoe
authors.forename: David E
authors.initials: DE
authors.lastname: Arnot
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood / Q000276:immunology; D000953:Antigens, Protozoan / Q000097:blood / Q000187:drug effects / Q000276:immunology; D000954:Antigens, Surface / Q000097:blood / Q000187:drug effects / Q000276:immunology; D002417:Cattle; D002448:Cell Adhesion; D002809:Chondroitin Sulfates / Q000378:metabolism*; D000939:Epitopes / Q000097:blood / Q000187:drug effects / Q000276:immunology; D004912:Erythrocytes / Q000276:immunology / Q000469:parasitology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D016778:Malaria, Falciparum / Q000097:blood / Q000276:immunology*; D008297:Male; D010963:Plasmodium falciparum / Q000276:immunology* / Q000502:physiology; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000097:blood / Q000276:immunology*; D015800:Protozoan Proteins / Q000097:blood / Q000187:drug effects / Q000276:immunology*; D014357:Trypsin / Q000494:pharmacology
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D000939:Epitopes; D007074:Immunoglobulin G; D015800:Protozoan Proteins; C085731:erythrocyte membrane protein 1, Plasmodium falciparum; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates; D014357:Trypsin