title: Analysis of intra-hepatic peptide-specific cell recruitment in mice immunised with Plasmodium falciparum antigens.
abstract: The liver stage of Plasmodium spp. now appears as a relevant target of immune effectors triggered by the so-called "anti-sporozoite" vaccine. Since the monitoring of immune responses at the systemic level may not faithfully reflect the local protective mechanisms, the aim of the present work was to set up a model to study the local intra-hepatic cellular responses and to compare these with the peripheral immune responses. This was achieved by intra-portal delivery of epitopic peptides, i.e. peptides containing B and T cell epitopes, which were coated onto the surface of polystyrene microbeads. The peptide-coated beads presumably mimic the hepatic schizont, and when distinct peptides are administered separately, this method of delivery allows us to decipher the immune responses resulting in mice immunised with recombinant proteins spanning several such epitopes. Using the P. falciparum liver stage antigen-3 (LSA3) molecule, which can induce protection against a sporozoite challenge, our results show that 25-microm microbeads could easily access the liver parenchyma by intra-portal injection and were distributed evenly in the liver. Also, LSA3-derived synthetic peptides coated onto microbeads initiated specific cell recruitment within 6 h. Depending on the LSA3 peptide used, the infiltrates induced differed in size, with the strongest cell recruitment obtained using nonrepeat II peptide (NR2)-coated microbeads with a mean leukocyte number of 79 per granuloma. Immunohistological studies of liver sections revealed that, irrespective of the delivered peptide, cells infiltrating the liver towards microbeads were mainly CD3(+) T lymphocytes, both CD4(+) (70 to 80%) and CD8(+) (20 to 30%) subtypes, macrophages and dendritic cells. Cells infiltrating the granuloma had features of activated cells, with evidence of VLA-4 cell-surface expression, and production of IFN-gamma and IL-4. Analysis of the peripheral B and T-cell responses in the same animals revealed that, whereas the local responses were directed mainly towards NR2 and repeat peptides (RE), the peripheral T-cell response to these peptides was weak and infrequent, although antibody production was high.
authors.affiliation: Unite de Parasitologie Médicale, Institut Pasteur, 25 & 28 Rue du Docteur Roux, 75724 cedex 15, Paris, France.
authors.forename: Armelle
authors.initials: A
authors.lastname: Hebert
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Sauzet
authors.forename: Mai
authors.initials: M
authors.lastname: Lebastard
authors.forename: Marie-Noëlle
authors.initials: MN
authors.lastname: Ungeheuer
authors.forename: Patrick
authors.initials: P
authors.lastname: Ave
authors.forename: Michel
authors.initials: M
authors.lastname: Huerre
authors.forename: Pierre
authors.initials: P
authors.lastname: Druilhe
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000008:administration & dosage / Q000235:genetics; D006099:Granuloma / Q000276:immunology / Q000473:pathology; D007114:Immunization; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000473:pathology; D008099:Liver / Q000276:immunology* / Q000469:parasitology / Q000473:pathology; D016778:Malaria, Falciparum / Q000276:immunology / Q000469:parasitology / Q000473:pathology; D051379:Mice; D008809:Mice, Inbred C3H; D008863:Microspheres; D008969:Molecular Sequence Data; D010316:Particle Size; D010446:Peptide Fragments / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000473:pathology
chemical_list: D000953:Antigens, Protozoan; D010446:Peptide Fragments; C067302:liver stage-specific antigen, Plasmodium