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Assessment of Balamuthia mandrillaris-specific serum antibody concentrations by flow cytometry

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Abstract

A flow cytometry (fluorescence-activated cell sorter)-based assay was adapted to detect and quantify antibodies to Balamuthia mandrillaris, a causative agent of fatal amoebic encephalitis (BAE), and to Acanthamoeba species. With sera from BAE patients for positive and a group of inconspicuous volunteers for negative reference, most of the 237 sera from random blood donors, patients with atypical encephalitis, atypical pneumonitis, visceral amoebiasis and toxoplasmosis and from subjects working with primates and other mammals were rated negative, 19% elevated and of these 2% high. In comparison, 23 of 25 West Africans living in rural areas revealed elevated, of these 15 high, and one very high B. mandrillaris-binding antibody titers, the latter well in the range of BAE patients. To date, none of the tested individuals have developed symptoms indicative of BAE. Criss-cross analysis with rabbit hyper immune sera against B. mandrillaris, Acanthamoeba comandoni (group 1), Acanthamoeba castellanii (group 2) and Acanthamoeba lenticulata (group 3) confirmed that cross-reactivity between B. mandrillaris and Acanthamoeba sp. is negligible while accentuating antigenic differences between the three morphological groups of Acanthamoeba.

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Abbreviations

BAE:

Balamuthia amoebic encephalitis

B.m.-Ab:

B. mandrillaris-binding antibody

CNS:

central nervous system

CSF:

cerebrospinal fluid

FB:

FACS buffer

FACS:

fluorescence-activated cell sorter

Ig:

immunoglobulin

IIF:

indirect immunofluorescence

MCSM:

modified Chang’s special medium

PBS:

phosphate-buffered saline

SD:

standard deviation

Sn:

supernatant

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Acknowledgements

We are very grateful to Jörg Blessmann, Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany; Rosemarie Elger, Health Services, Free University of Berlin; Klaus Eulenberg, Zoological Garden Leipzig; Thomas Illmer and Gert Höffken, Carl Gustav Carus Clinic, Technical University of Dresden; Fabian H. Leendertz, Emerging Zoonotic Diseases Group, Robert Koch Institute, Berlin; Oliver Liesenfeld, Institute for Medical Microbiology and Immunology of Infections, Free University of Berlin; Andreas Roth, Institute for Microbiology and Immunology, Medical Center Zehlendorf-Heckeshorn, Berlin; Christine Scheuch, Haema Blood Bank, Berlin; Eva Schielke, Neurological Clinic and Polyclinic, Humboldt University, Berlin; Frederick L. Schuster, California Department of Health Services, Richmond, USA; Govinda S. Visvesvara, Centers for Disease Control and Prevention, Atlanta, USA; and to all individual donors for providing us with serum samples. We thank Ulrike Laube for expert technical assistance.

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  1. Cellular Immunology Unit P22, Robert Koch Institute, Nordufer 20, 13353, Berlin, Germany

    Albrecht F. Kiderlen, Elke Radam & Phiroze S. Tata

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  1. Albrecht F. Kiderlen
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  2. Elke Radam
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  3. Phiroze S. Tata
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Correspondence to Albrecht F. Kiderlen.

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Kiderlen, A.F., Radam, E. & Tata, P.S. Assessment of Balamuthia mandrillaris-specific serum antibody concentrations by flow cytometry. Parasitol Res 104, 663–670 (2009). https://doi.org/10.1007/s00436-008-1243-6

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  • DOI: https://doi.org/10.1007/s00436-008-1243-6

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