Warning: mkdir(): Permission denied in /home/virtual/lib/view_data.php on line 81

Warning: fopen(upload/ip_log/ip_log_2024-03.txt): failed to open stream: No such file or directory in /home/virtual/lib/view_data.php on line 83

Warning: fwrite() expects parameter 1 to be resource, boolean given in /home/virtual/lib/view_data.php on line 84
The production and characterization of anti-Naegleria fowleri monoclonal antibodies
| Home | E-Submission | Sitemap | Contact us |  
top_img
Korean J Parasito Search

CLOSE

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

References
1. Alderete JF, Pearlman E. Pathogenic Trichomonas vaginalis cytotoxicity to cell culture monolayers. Br J Vener Dis 1984;60(2):99–105.
 
2. Bickle QD, Andrews BJ, Taylor MG. Schistosoma mansoni: characterization of two protective monoclonal antibodies. Parasite Immunol 1986;8(1):95–107.
  
3. Burgess DE. Tritrichomonas foetus: preparation of monoclonal antibodies with effector function. Exp Parasitol 1986;62(2):266–274.
  
4. Epstein N, Miller LH, Kaushel DC, Udeinya IJ, Rener J, Howard RJ, Asofsky R, Aikawa M, Hess RL. Monoclonal antibodies against a specific surface determinant on malarial (Plasmodium knowlesi) merozoites block erythrocyte invasion. J Immunol 1981;127(1):212–217.
 
5. Ferrante A, Thong YH. Antibody induced capping and endocytosis of surface antigens in Naegleria fowleri. Int J Parasitol 1979;9(6):599–601.
  
6. Haggerty RM, John DT. Serum agglutination and immunoglobulin levels of mice infected with Naegleria fowleri. J Protozool 1982;29(1):117–122.
 
7. Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975;256(5517):495–497.
  
8. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227(5259):680–685.
  
9. Lallinger GJ, Reiner SL, Cooke DW, Toffaletti DL, Perfect JR, Granger DL, Durack DT. Efficacy of immune therapy in early experimental Naegleria fowleri meningitis. Infect Immun 1987;55(5):1289–1293.
 
10. Lee YW, et al. Yonsei J Med Sci 1986;19(2):358–369.
11. Littlefield JW. Selection Of Hybrids From Matings Of Fibroblasts In Vitro And Their Presumed Recombinants. Science 1964;145:709–710.
  
12. Nash TE, Aggarwal A. Cytotoxicity of monoclonal antibodies to a subset of Giardia isolates. J Immunol 1986;136(7):2628–2632.
 
13. Ringsted J, Jager BV, Suk D, Visvesvara GS. Probable acanthamoeba meningoencephalitis in a Korean child. Am J Clin Pathol 1976;66(4):723–730.
 
14. Seidel JS, Harmatz P, Visvesvara GS, Cohen A, Edwards J, Turner J. Successful treatment of primary amebic meningoencephalitis. N Engl J Med 1982;306(6):346–348.
  
15. Thompson JP, Crandall RB, Crandall CA, Neilson JT. Clearance of microfilariae of Dipetalonema viteae in CBA/N and CBA/H mice. J Parasitol 1979;65(6):966–969.
  
16. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 1979;76(9):4350–4354.
  
17. Visvesvara GS, Healy GR. Comparative antigenic analysis of pathogenic and free-living Naegleria species by the gel diffusion and immunoelectrophoresis techniques. Infect Immun 1975;11(1):95–108.
 
18. Weiss N. Monoclonal antibodies as investigative tools in onchocerciasis. Rev Infect Dis 1985;7(6):826–830.
  
19. Willaert E. Ann Soc Belg Med Trop 1971;51:701–708.
Editorial Office
Department of Molecular Parasitology, Samsung Medical Center, School of Medicine, Sungkyunkwan University,
2066 Seobu-ro, Jangan-gu, Suwon 16419, Gyeonggi-do, Korea.
Tel: +82-31-299-6251   FAX: +82-1-299-6269   E-mail: kjp.editor@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © 2024 by The Korean Society for Parasitology and Tropical Medicine.     Developed in M2PI