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Immunological characterizations of a cloned 13.1-kilodalton protein from pathogenic Naegleria fowleri.

Authors
Cho, MS; Jung, SY; Park, S; Kim, KH; Kim, HI; Sohn, S; Kim, HJ; Im, KI; Shin, HJ
Citation
Clinical and diagnostic laboratory immunology, 10(5):954-959, 2003
Journal Title
Clinical and diagnostic laboratory immunology
ISSN
1071-412X1098-6588
Abstract
We previously cloned an antigenic gene (named nfa1) from a cDNA library of Naegleria fowleri by immunoscreening. The nfa1 gene had a coding nucleotide sequence consisting of 357 bases and produced a recombinant 13.1-kDa protein (Nfa1). In this study, to get more information regarding the recombinant Nfa1 protein (rNfa1), we produced an anti-Nfa1 polyclonal antibody from mice immunized with rNfa1 and used a peroxidase staining method to carry out immunocytochemistry experiments. In addition, we observed the effect of the presence of an anti-Nfa1 antibody on the in vitro cytotoxicity of N. fowleri against Chinese hamster ovary (CHO) cells. Trophozoites of N. fowleri in cultivation reacted strongly with a peroxidase-labeled anti-Nfa1 antibody. In inflammatory and necrotic regions of brain tissue infected with N. fowleri, labeled trophozoites that were stained brown were also observed. When examined using a transmission electron microscope, the Nfa1 protein showed pseudopodium-specific immunolocalization on a trophozoite of N. fowleri. When examined using a light microscope, CHO cells grown in cocultures with N. fowleri trophozoites (group I) for 48 h showed morphologically severe destruction but CHO cells grown in cocultures with N. fowleri trophozoites and an anti-Nfa1 polyclonal antibody (group II) showed less destruction. The results of a lactate dehydrogenase release assay showed that group I CHO cells exhibited 81% cytotoxicity and group II CHO cells exhibited 13.8% cytotoxicity.
MeSH terms
Amebiasis/immunologyAnimalsAntibodies, Protozoan/immunology*Antigens, Protozoan/geneticsAntigens, Protozoan/immunology*Blotting, WesternBrain/microbiologyCHO CellsCloning, MolecularCoculture TechniquesCricetinaeCytotoxicity Tests, ImmunologicCytotoxicity, Immunologic*Electrophoresis, Polyacrylamide GelEnzyme-Linked Immunosorbent AssayFemaleImmunohistochemistryMiceMicroscopy, ElectronNaegleria fowleri/immunology*Naegleria fowleri/ultrastructurePolymerase Chain ReactionProtozoan Proteins/geneticsProtozoan Proteins/immunology*Recombinant Proteins/immunology
PMID
12965933
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
Journal Papers > School of Medicine / Graduate School of Medicine > Gastroenterology
AJOU Authors
박, 선김, 경민김, 형일손, 성향신, 호준
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