The production and characterization of anti-<i>Naegleria fowleri</i> monoclonal antibodies

, Ryu, J S; , Im, K I

doi:1992.30.1.33

Korean J Parasito > Volume 30(1):1992 > Article

Original Article


Korean J Parasitol. 1992 Mar;30(1):33-41. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1992.30.1.33
Copyright © 1992 by The Korean Society for Parasitology


The production and characterization of anti-Naegleria fowleri monoclonal antibodies
J S Ryu and K I Im^*
Department of Parasitology, College of Medicine, Hanyang University 133-791, Seoul, Korea.
^*Department of Parasitology, College of Medicine, Yonsei University, Seoul 120-752, Korea.


Abstract
Naegleria fowleri, a free-living amoeba commonly found in moist soil and fresh water, enters the body via the nasal mucosa and migrates along the olfactory nerve to the brain, where it causes acute amoebic meningoencephalitis. In the present study 7 clones secreting monoclonal antibodies (McAbs) against N. fowleri were produced and the effector function of them was investigated. Their isotypes were IgG1 (Nf 1, Nf 154), IgG3 (Nf 137) and IgA (Nf 1, Nf 2, Nf 256, Nf 279). Five McAbs (McAb Nf 2, Nf 279, Nf 27, Nf 154, Nf 137) were specific for N. fowleri by ELISA and recognized the antigenic determinants located on the trophozoite surface by IFAT and immunoperoxidase stain. These five McAbs had capacity to agglutinate N. fowleri trophozoites and inhibited the growth of the amoeba in culture medium. McAb Nf 2 inhibited proliferation of trophozoites in vitro significantly. Also the cytotoxicity of N. fowleri against CHO cell was reduced in the presence of McAb Nf 2 and McAb Nf 154. From these results McAb Nf 2 was confirmed to weaken the virulence of the amoeba among 7 screened McAbs.

Figures


	Fig. 1 (A) Indirect immunofluorescence findings of N. fowleri trophozoites incubated with McAb Nf 2 showing fluorescence in localized area of cell membrane. (B) McAb Nf 1 showing diffuse fluorescence on the cell membrane.


	Fig. 2 (A) EM of N. fowleri trophozoites incubated with culture supernatant without McAb. Peroxidase activity was demonstrated on mitochondria only. (B) EM of N. fowleri trophozoites incubated with McAb Nf 154. Peroxidase activity was observed over the amoeba surface membrane and mitochondria. N; nucleus, M; mitochondria, V; vacuole


	Fig. 3 Enzyme-linked immunoelectrotransfer blotting of N. fowleri lysate reacted with various monoclonal antibodies. M.W. protein standards(1), Nf 137(2), Nf 279(3) and Nf 154(4)

Tables


	Table 1 Isotype of monoclonal antibodies


	Table 2 Cross-reactivity of anti-N. fowleri monoclonal antibodies with free-living amoeba, measured by ELISA


	Table 3 Agglutination of N. fowleri trophozoites by anti-N. fowleri monoclonal antibodies


	Table 4 Effect of monoclonal antibodies on in vitro proliferation of N. fowleri


	Table 5 Effect of monoclonal antibodies on cytotoxicity against Chinese hamster ovary cells by N. fowleri


	Table 6 Summary of characteristics of 7 anti-N. fowleri monoclonal antibodies

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