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Accessibility and distribution of intraerythrocytic antigens ofPlasmodium-infected erythrocytes following mild glutaraldehyde fixation and detergent extraction

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Abstract

Malarial antigens on the surface of infected erythrocytes have been described by many investigators. However, few of these antigens have been unambiguously demonstrated to be exposed on the surface of erythrocytes. This study demonstrates that mild glutaraldehyde fixation results in the cytoplasmic face of the host membrane becoming accessible to antibody under conditions that normally do not expose the cytoplasmic face of uninfected erythrocytes. These results indicate that caution should be used in interpreting data on the membrane disposition of malarial antigens. Detergent extraction of the glutaraldehyde-fixed erythrocytes results in an increased permeabilization such that malarial antigens on the parasite surface and within the cytoplasm of the infected erythrocyte are accessible to antibody. The accessibility of these antigens was demonstrated by both immunofluorescence and two-color flow cytometry. The antigens within the host cytoplasm were not diffuse but associated with patchy aggregates. Analysis of the antigens associated with the cytoplasmic aggregates by immunoelectron microscopy indicated that they were not associated with membrane-bound compartments. The fixation and permeabilization protocol described herein will have useful applications for the characterization and analysis of malarial antigens.

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Abbreviations

Ag:

antigen

FITC:

fluorescein isothiocyanate

HBSS:

Hanks' balanced salt solution

mAb:

monoclonal antibody

MSP1:

merozoite surface protein 1

PBS:

phosphate-buffered saline

PfHRPII:

Plasmodium falciparum histidine-rich protein II

RESA:

ring-stage erythrocyte surface antigen

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Authors and Affiliations

  1. Department of Tropical Medicine, Tulane University School of Public Health and Tropical Medicine, 70112, New Orleans, LA, USA

    Mark F. Wiser & Lynn V. V. Faur

  2. Interdisciplinary Program in Molecular and Cell Biology, Tulane University, 70112, New Orleans, LA, USA

    Mark F. Wiser, H. Norbert Lanners, Mary Kelly & Russell B. Wilson

  3. Tulane Regional Primate Research Center, 70433, Covington, LA, USA

    H. Norbert Lanners

  4. Department of Pathology and Laboratory Medicine, Tulane University School of Medicine, 70112, New Orleans, LA, USA

    Russell B. Wilson

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  1. Mark F. Wiser
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  2. Lynn V. V. Faur
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  3. H. Norbert Lanners
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  4. Mary Kelly
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  5. Russell B. Wilson
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Wiser, M.F., Faur, L.V.V., Lanners, H.N. et al. Accessibility and distribution of intraerythrocytic antigens ofPlasmodium-infected erythrocytes following mild glutaraldehyde fixation and detergent extraction. Parasitol Res 79, 579–586 (1993). https://doi.org/10.1007/BF00932242

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  • DOI: https://doi.org/10.1007/BF00932242

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