title: Plasmodium falciparum synthetic LbL microparticle vaccine elicits protective neutralizing antibody and parasite-specific cellular immune responses.
abstract: Epitopes of the circumsporozoite (CS) protein of Plasmodium falciparum, the most pathogenic species of the malaria parasite, have been shown to elicit protective immunity in experimental animals and human volunteers. The mechanisms of immunity include parasite-neutralizing antibodies that can inhibit parasite motility in the skin at the site of infection and in the bloodstream during transit to the hepatocyte host cell and also block interaction with host cell receptors on hepatocytes. In addition, specific CD4+ and CD8+ cellular mechanisms target the intracellular hepatic forms, thus preventing release of erythrocytic stage parasites from the infected hepatocyte and the ensuing blood stage cycle responsible for clinical disease. An innovative method for producing particle vaccines, layer-by-layer (LbL) fabrication of polypeptide films on solid CaCO3 cores, was used to produce synthetic malaria vaccines containing a tri-epitope CS peptide T1BT comprising the antibody epitope of the CS repeat region (B) and two T-cell epitopes, the highly conserved T1 epitope and the universal epitope T. Mice immunized with microparticles loaded with T1BT peptide developed parasite-neutralizing antibodies and malaria-specific T-cell responses including cytotoxic effector T-cells. Protection from liver stage infection following challenge with live sporozoites from infected mosquitoes correlated with neutralizing antibody levels. Although some immunized mice with low or undetectable neutralizing antibodies were also protected, depletion of T-cells prior to challenge resulted in the majority of mice remaining resistant to challenge. In addition, mice immunized with microparticles bearing only T-cell epitopes were not protected, demonstrating that cellular immunity alone was not sufficient for protective immunity. Although the microparticles without adjuvant were immunogenic and protective, a simple modification with the lipopeptide TLR2 agonist Pam3Cys increased the potency and efficacy of the LbL vaccine candidate. This study demonstrates the potential of LbL particles as promising malaria vaccine candidates using the T1BT epitopes from the P. falciparum CS protein.
authors.affiliation: Artificial Cell Technologies, Inc., 5 Science Park, Suite 13, New Haven, CT 06511, United States. jpowell@artificialcelltech.com
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Powell
authors.forename: Jie
authors.initials: J
authors.lastname: Tang
authors.forename: Mary E
authors.initials: ME
authors.lastname: Derome
authors.forename: Robert A
authors.initials: RA
authors.lastname: Mitchell
authors.forename: Andrea
authors.initials: A
authors.lastname: Jacobs
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Deng
authors.forename: Naveen
authors.initials: N
authors.lastname: Palath
authors.forename: Edwin
authors.initials: E
authors.lastname: Cardenas
authors.forename: James G
authors.initials: JG
authors.lastname: Boyd
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Nardin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000913:Antibodies, Protozoan / Q000097:blood / Q000276:immunology; D009033:Culicidae / Q000469:parasitology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005260:Female; D007111:Immunity, Cellular / Q000276:immunology*; D008288:Malaria / Q000276:immunology* / Q000517:prevention & control*; D017780:Malaria Vaccines / Q000138:chemical synthesis* / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010271:Parasites / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D010963:Plasmodium falciparum / Q000276:immunology* / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000276:immunology; D034101:Sporozoites / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000276:immunology*
chemical_list: D057134:Antibodies, Neutralizing; D000913:Antibodies, Protozoan; D018984:Epitopes, T-Lymphocyte; D017780:Malaria Vaccines; D010455:Peptides; D015800:Protozoan Proteins; C516102:T1BT peptide; D014614:Vaccines, Synthetic; C041595:circumsporozoite protein, Protozoan