Immunological Characterizations of a Cloned 13.1-Kilodalton Protein from Pathogenic
            <i>Naegleria fowleri</i>

Cho, Myoung-Soo; Jung, Suk‐Yul; Park, Sun; Kim, Kyongmin; Kim, Hyung-Il; Sohn, Seonghyang; Kim, Han-Jip; Im, Kyongok; Shin, Ho‐Joon

doi:10.1128/cdli.10.5.954-959.2003

Cited by 20 publications

(29 citation statements)

References 23 publications

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“…The nfa1 gene, which is involved with amoebic pseudopodial activity and especially with food cup formation, plays an important role in the pathogenicity of N. fowleri infection (13,14). Moreover, an anti-Nfa1 antibody caused a decrease in the cytotoxicity of N. fowleri against target cells (15).…”

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confidence: 99%

“…Mice immunized with an intraperitoneal inoculation with live or killed trophozoites of N. fowleri showed variable levels of partial protective immunity (25). According to our previous studies, the nfa1 gene may be a proper candidate for DNA vaccination against N. fowleri infection (12)(13)(14)26). Based on these findings, to evaluate the effect of our lentiviral vector systems expressing the nfa1 gene in the in vivo mouse model, vaccinated mice were tested for the development of specific immunity against N. fowleri infection, measured by humoral and cellular immune responses and by survival rates.…”

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confidence: 99%

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Vaccination with Lentiviral Vector Expressing thenfa1Gene Confers a Protective Immune Response to Mice Infected with Naegleria fowleri

Kim

Sohn

Lee

et al. 2013

Clin. Vaccine Immunol.

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confidence: 99%

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confidence: 99%

Vaccination with Lentiviral Vector Expressing thenfa1Gene Confers a Protective Immune Response to Mice Infected with Naegleria fowleri

Kim

Sohn

Lee

et al. 2013

Clin. Vaccine Immunol.

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“…We previously reported that the nfa1 gene cloned from pathogenic N. fowleri encodes a 13.1-kDa antigenic protein (23). This nfa1 gene consists of 360 bp, and the Nfa1 gene product is localized to the pseudopodia of trophozoites, which may be related to the movement and pathogenicity of N. fowleri, as determined by transmission electron microscopy and immunocytochemistry using an anti-Nfa1 antibody obtained by immunizing mice with recombinant Nfa1 (rNfa1) protein derived from the N. fowleri nfa1 gene (2,10,23). In addition, anti-Nfa1 antibody treatment reduced the cytotoxicity of N. fowleri for Chinese hamster ovary (CHO) cells in a dose-dependent manner, supporting the notion that the nfa1 gene may be related to the pathogenicity of Naegleria spp.…”

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confidence: 99%

“…In addition, anti-Nfa1 antibody treatment reduced the cytotoxicity of N. fowleri for Chinese hamster ovary (CHO) cells in a dose-dependent manner, supporting the notion that the nfa1 gene may be related to the pathogenicity of Naegleria spp. (2,10).…”

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Expression of the nfa1 Gene Cloned from Pathogenic Naegleria fowleri in Nonpathogenic N. gruberi Enhances Cytotoxicity against CHO Target Cells In Vitro

et al. 2005

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The pathogenic amoeba Naegleria fowleri has a 360-bp nfa1 gene that encodes the Nfa1 protein (13.1 kDa), which is located in the pseudopodia of the amoeba, and an anti-Nfa1 antibody reduces N. fowleri-induced mammalian-cell cytotoxicity in vitro. In contrast, an anti-Nfa1 antibody cannot detect Nfa1 protein expression in the nonpathogenic amoeba Naegleria gruberi, which also possesses the nfa1 gene. In the present study, the nfa1 gene cloned from pathogenic N. fowleri was transfected into nonpathogenic N. gruberi to determine whether it was related to pathogenicity. The nfa1 gene was initially inserted into a eukaryotic transfection vector, pEGFP-C2, containing a cytomegalovirus promoter and the green fluorescent protein (GFP) gene, and was designed as pEGFP-C2/nfa1UTR (nfa1UTR contains 5 upstream regions, the nfa1 open reading frame, and 3 downstream regions). After transfection, the green fluorescence was observed in the cytoplasm of N. gruberi trophozoites. These transfectants were preserved for more than 9 months after selection. The transfected nfa1 gene was observed by PCR using nfa1-and vector-specific primers in the genomic DNA of N. gruberi transfected with the pEGFP-C2/nfa1UTR vector. In addition, the nfa1 and GFP genes were identified by reverse transcription-PCR in transgenic N. gruberi. The Nfa1 protein expressed in transgenic N. gruberi was identified as a 13.1-kDa band by Western blotting using an anti-Nfa1 antibody. Finally, N. gruberi transfected with the pEGFP-C2/nfa1UTR vector was found to have enhanced cytotoxicity against CHO cells compared with naïve N. gruberi.

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Production of Nfa1-specific monoclonal antibodies that influences the in vitro cytotoxicity of Naegleria fowleri trophozoites on microglial cells

et al. 2007

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Naegleria fowleri, agent of fatal primary amoebic meningoencephalitis, appears to induce cytotoxicity mechanically through its contact with the cell. The nfa1 gene cloned from a cDNA library of pathogenic N. fowleri by immunoscreening consists of 360 bp and expresses a 13.1-kDa recombinant protein (rNfa1) that demonstrated localization in the pseudopodia when examined using immunocytochemistry. To study the mechanisms involved in N. fowleri cytotoxicity, we developed a large volume of rNfa1-specific monoclonal antibody (McAb) against a 17-kDa His-tag fusion rNfa1 protein using a cell fusion technique. We established eight McAb-producing hybridoma cells. The antibodies were all immunoglobulin G2b and reacted strongly with a 17-kDa band representing the rNfa1 fusion protein in Western blotting, demonstrating immunoreactivity to the Nfa1 protein in pseudopodia (especially in the food cups) of N. fowleri trophozoites. A 51 Cr-release assay indicated N. fowleri cytotoxicity by demonstrating that it eliminated 37.8, 60.6, and 98.8% of the target (microglial) cells 6, 12, and 24 h after coincubation, respectively. When an anti-Nfa1 McAb was added to the coculture system, N. fowleri cytotoxicity decreased to 29.8, 44.1, and 66.3%, respectively.

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

Cited by 20 publications

References 23 publications

Vaccination with Lentiviral Vector Expressing thenfa1Gene Confers a Protective Immune Response to Mice Infected with Naegleria fowleri

Vaccination with Lentiviral Vector Expressing thenfa1Gene Confers a Protective Immune Response to Mice Infected with Naegleria fowleri

Expression of the nfa1 Gene Cloned from Pathogenic Naegleria fowleri in Nonpathogenic N. gruberi Enhances Cytotoxicity against CHO Target Cells In Vitro

Production of Nfa1-specific monoclonal antibodies that influences the in vitro cytotoxicity of Naegleria fowleri trophozoites on microglial cells

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