Parasitology Research

, Volume 102, Issue 5, pp 997–1000 | Cite as

Cytopathogenesis of Naegleria fowleri Thai strains for cultured human neuroblastoma cells

  • Supathra Tiewcharoen
  • Nat Malainual
  • Virach Junnu
  • Pruksawan Chetanachan
  • Jundee Rabablert
Original Paper


The aim of this study is to evaluate cellular interaction between free-living amoebae Naegleria fowleri strains and mammalian target cells in vitro. Two Thai strains of N. fowleri; Khon Kaen strain from the environment and Siriraj strain from the patient’s cerebrospinal fluid and the Center of Disease Control VO 3081 strain from Atlanta (US) were studied. Human neuroblastoma (SK-N-MC) and African Green monkey Kidney (Vero) cells were used as target cells. Each cell line was inoculated with each strain of N. fowleri at a ratio of 1:1 and observed for 7 days. The uninoculated target cells and each strain of N. fowleri were used as control. The numbers of the challenged and unchallenged cells as well as the free-living amoebae were counted three times by trypan blue exclusion method. The inoculation began when the amoebae attached to the cell membrane and ingested the target cells. In this study, extensive cytopathogenesis with many floating inoculated cells and abundant number of amoebae were observed. The destruction pattern of both inoculated SK-N-MC and Vero target cells were similar. Interestingly, SK-N-MC was more susceptible to N. fowleri strains than the Vero cell. In addition, N. fowleri Siriraj strain showed the highest destruction pattern for each target cell. Our findings suggest that the SK-N-MC should be used as a base model for studying the neuropathogenesis in primary amoebic meningoencephalitis patients.


Cytopathogenesis Naegleria fowleri Thai strains Human neuroblastoma cells 



This work was partially funded by the Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand in 2005–2006.


  1. Biedler JL, Helson L, Spengler BA (1973) Morphology and growth, tumorigenicity, and cytogenetics of human neuroblastoma cells in continuous culture. Cancer Res 3:2643–2652Google Scholar
  2. De Jonckheere JF (2002) A century of research on the amoeboflagellate genus Naegleria. Acta Protozool 41:309–342Google Scholar
  3. Fulford DE, Marciano-Cabral F (1986) Cytopathic activity of Naegleria fowleri cell-free extract. J Protozool 33:498–450PubMedGoogle Scholar
  4. Jeong SR, Lee SC, Song KJ, Park S, Kim K, Kwon MH, Im Ki, Shin HJ (2005) Expression on the nfa1 gene cloned from pathogenic Naegleria fowleri in nonpathogenic N. gluberi enhances cytotoxicity against CHO target cells in vitro. Infect Immun 73:4098–4105PubMedCrossRefGoogle Scholar
  5. John DT, John RA (1994a) Use of Vero-cell culture to assess cytopathogenicity of Naegleria species. Proc Okla Acad 74:17–20Google Scholar
  6. John DT, John RA (1994b) Enhancement of virulence of Naegleria fowleri by growth in Vero-cell culture. J Parasitol 80:149–150PubMedCrossRefGoogle Scholar
  7. Marciano-Cabral F, John DT (1983) Cytopathogenicity of Naegleria fowleri for rat neuroblastoma cell cultures: scanning electron microscopy study. Infect Immun 40:1214–1217PubMedGoogle Scholar
  8. Nelson EC, Jones MM (1970) Culture isolation of agents of primary amebic meningoencephalitis. J Parasitol 56:248CrossRefGoogle Scholar
  9. Song KJ, Song KH, Na BK, Kim JH,Kwon D, Park S, Pak JH, Im KII, Shin HJ (2007) Molecular cloning and characterization of a cytosolic heat shock protein 70 from Naegleria fowleri. Parasitol Res 100:1083–1089 DOI  1007/s00436–006–0404–8 Google Scholar
  10. Tiewcharoen S, Komalamisara N, Junnu V (2004) Naegleria fowleri in Thailand 2003. Southeast Asian J Trop Med Public Health 35:24–31Google Scholar
  11. Yum H-C, Park S-J, Kong H-H, Chung D-I (2002) Isolation of genes induced in Naegleria fowleri during mouse brain passage. Eur J Protisol 38:105–111CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Supathra Tiewcharoen
    • 1
  • Nat Malainual
    • 1
  • Virach Junnu
    • 1
  • Pruksawan Chetanachan
    • 3
  • Jundee Rabablert
    • 2
  1. 1.Department of Parasitology, Faculty of MedicineSiriraj Hospital, Mahidol UniversityBangkokThailand
  2. 2.Department of Biology, Faculty of ScienceSilpakorn UniversityNakorn PhathomThailand
  3. 3.Department of Medical ScienceNational Institute of HealthNonthaburiThailand

Personalised recommendations