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Clinical and Diagnostic Laboratory Immunology, September 2003, p. 954-959, Vol. 10, No. 5
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.5.954-959.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Immunological Characterizations of a Cloned 13.1-Kilodalton Protein from Pathogenic Naegleria fowleri

Myoung-Soo Cho,1 Suk-Yul Jung,1 Sun Park,1 Kyongmin Hwang Kim,1 Hyung-Il Kim,1 Seonghyang Sohn,2 Han-Jip Kim,3 Kyung-Il Im,4 and Ho-Joon Shin1*

Department of Microbiology, School of Medicine,1 Laboratory of Cell Biology, Institute for Medical Science,2 Division of Natural Science, Ajou University, Suwon 442-749,3 Department of Parasitology, College of Medicine, Yonsei University, Seoul 121-752, Korea4

Received 5 March 2003/ Returned for modification 5 May 2003/ Accepted 5 June 2003

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.

* Corresponding author. Mailing address: Department of Microbiology, Ajou University School of Medicine, Suwon 442-749, Korea. Phone: 82 31 219-5076. Fax: 82 31 219-5079. E-mail: hjshin{at}ajou.ac.kr.

Clinical and Diagnostic Laboratory Immunology, September 2003, p. 954-959, Vol. 10, No. 5
1071-412X/03/$08.00+0     DOI: 10.1128/CDLI.10.5.954-959.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kang, S.-Y., Song, K.-J., Jeong, S.-R., Kim, J.-H., Park, S., Kim, K., Kwon, M.-H., Shin, H.-J. (2005). Role of the Nfa1 Protein in Pathogenic Naegleria fowleri Cocultured with CHO Target Cells. CVI 12: 873-876 [Abstract] [Full Text]  
  • Jeong, S.-R., Lee, S.-C., Song, K.-J., Park, S., Kim, K., Kwon, M.-H., Im, K.-i., Shin, H.-J. (2005). Expression of the nfa1 Gene Cloned from Pathogenic Naegleria fowleri in Nonpathogenic N. gruberi Enhances Cytotoxicity against CHO Target Cells In Vitro. Infect. Immun. 73: 4098-4105 [Abstract] [Full Text]  


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