Identification of a Naegleria fowleri membrane protein reactive with anti-human CD59 antibody
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High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum
Published on: November 23, 2013
Angela E Fritzinger1, Denise M Toney, Rebecca C MacLean
1Department of Microbiology and Immunology, Virginia Commonwealth University School of Medicine, Sanger Hall, 1101 E. Marshall Street, P.O. Box 980678, Richmond, VA 23298-0678, USA.
Abstract:
Naegleria fowleri, the causative agent of primary amebic meningoencephalitis, is resistant to complement lysis. The presence of a complement regulatory protein on the surface of N. fowleri was investigated. Southern blot and Northern blot analyses demonstrated hybridization of a radiolabeled cDNA probe for CD59 to genomic DNA and RNA, respectively, from pathogenic N. fowleri. An 18-kDa immunoreactive protein was detected on the membrane of N. fowleri by Western immunoblot and immunofluorescence analyses with monoclonal antibodies for human CD59. Complement component C9 immunoprecipitated with the N. fowleri "CD59-like" protein from amebae incubated with normal human serum. In contrast, a gene or protein similar to CD59 was not detected in nonpathogenic, complement-sensitive N. gruberi amebae. Collectively, our studies suggest that a protein reactive with antibodies to human CD59 is present on the surface of N. fowleri amebae and may play a role in resistance to lysis by cytolytic proteins.
Insights
Naegleria fowleri, a pathogen causing primary amebic meningoencephalitis, may evade immune attack using a CD59-like protein. This protein appears to protect the ameba from complement-mediated lysis.
Area of Science:
- Immunology
- Microbiology
- Cell Biology
Background:
- Naegleria fowleri causes primary amebic meningoencephalitis and resists complement lysis.
- Understanding resistance mechanisms is crucial for treating this infection.
Purpose of the Study:
- To investigate the presence and role of complement regulatory proteins on Naegleria fowleri.
- To determine if N. fowleri utilizes proteins similar to human CD59 for immune evasion.
Main Methods:
- Southern and Northern blot analyses using a human CD59 cDNA probe.
- Western immunoblot and immunofluorescence assays with anti-CD59 antibodies.
- Immunoprecipitation of complement component C9.
Main Results:
- A CD59-like protein (18 kDa) was detected on pathogenic N. fowleri but not on nonpathogenic N. gruberi.
- This protein reacted with antibodies against human CD59.
- The N. fowleri CD59-like protein immunoprecipitated complement component C9.
Conclusions:
- Naegleria fowleri expresses a surface protein reactive with anti-CD59 antibodies.
- This CD59-like protein may contribute to N. fowleri's resistance to complement-mediated lysis.
- The findings suggest a novel immune evasion strategy for this pathogen.
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