title: Cytotoxic T cell reactivity and HLA-B35 binding of the variant Plasmodium falciparum circumsporozoite protein CD8+ CTL epitope in naturally exposed Kenyan adults.
abstract: In this study, we have investigated the extent of natural polymorphism in the CD8+ cytotoxic T lymphocyte (CTL) determinant (amino acids 368-390) of circumsporozoite (CS) protein of Plasmodium falciparum field isolates from a holoendemic region of Kenya, and determined how this variation affects the CTL reactivities in clinically immune adults and binding specificities to human histocompatibility leukocyte antigen (HLA)-B35. Among the eight variant sequences that were found in this region, four were new and not seen in parasites from other geographical regions. When synthetic peptides corresponding to the eight variants were used to test the presence of CTL response in different donors, a different spectrum of CTL reactivity to these variants was noticed. While CTL from some donors recognized the P1 sequence (the most prevalent type of sequence) but not P8 (another major variant), other donors showed a reverse pattern of reactivity. Although none of the donors was able to recognize all the variants, CTL responses to all the eight variant sequences were found in this population. An octamer peptide with P1 sequence KPKDELDY in this polymorphic determinant was known to bind HLA-B35. When we tested the effect of natural variation in this octamer sequence on HLA-B35 binding, it became evident that SP13 with D --> N substitution retained its binding specificity to HLA-B35. On the other hand, the SP12 octamer sequence which had two substitutions did not bind HLA-B35. The most interesting finding was the observation that a D --> G substitution at position 374 rescued the binding ability of SP14, which otherwise could not bind to this HLA molecule due to E --> Q amino acid substitution at position 372. To our knowledge, this is the first demonstration showing that a natural polymorphism can rescue the binding specificity to an HLA-class I molecule that was lost due to another natural amino acid substitution. Altogether, these results demonstrate that natural polymorphism in the CS protein affects both the CTL reactivity and the ability to bind to HLA-B35.
authors.affiliation: Division of Parasitic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30341-3724, USA. Kumar@FORM-NET.COM
authors.forename: V
authors.initials: V
authors.lastname: Udhayakumar
authors.forename: J M
authors.initials: JM
authors.lastname: Ongecha
authors.forename: Y P
authors.initials: YP
authors.lastname: Shi
authors.forename: M
authors.initials: M
authors.lastname: Aidoo
authors.forename: A S
authors.initials: AS
authors.lastname: Orago
authors.forename: A J
authors.initials: AJ
authors.lastname: Oloo
authors.forename: W A
authors.initials: WA
authors.lastname: Hawley
authors.forename: B L
authors.initials: BL
authors.lastname: Nahlen
authors.forename: S L
authors.initials: SL
authors.lastname: Hoffman
authors.forename: W R
authors.initials: WR
authors.lastname: Weiss
authors.forename: A A
authors.initials: AA
authors.lastname: Lal
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D016054:DNA, Protozoan / Q000235:genetics; D000939:Epitopes / Q000235:genetics / Q000378:metabolism; D014644:Genetic Variation; D015797:HLA-B35 Antigen / Q000378:metabolism*; D006801:Humans; D007630:Kenya; D016778:Malaria, Falciparum / Q000276:immunology* / Q000469:parasitology; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology*; D011485:Protein Binding; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
chemical_list: D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D000939:Epitopes; D015797:HLA-B35 Antigen; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan