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Phenotypic and functional characterization of earthworm coelomocyte subsets: Linking light scatter-based cell typing and imaging of the sorted populations.

Engelmann, Péter and Hayashi, Yuya and Bodó, Kornélia and Ernszt, Dávid and Némethné Somogyi, Ildikó and Steib, Anita and Orbán, József and Pollák, Edit and Nyitrai, Miklós and Németh, Péter and Molnár, László (2016) Phenotypic and functional characterization of earthworm coelomocyte subsets: Linking light scatter-based cell typing and imaging of the sorted populations. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY, 65. pp. 41-52. ISSN 0145-305X

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Abstract

Flow cytometry is a common approach to study invertebrate immune cells including earthworm coelomocytes. However, the link between light-scatter- and microscopy-based phenotyping remains obscured. Here we show, by means of light scatter-based cell sorting, both subpopulations (amoebocytes and eleocytes) can be physically isolated with good sort efficiency and purity confirmed by downstream morphological and cytochemical applications. Immunocytochemical analysis using anti-EFCC monoclonal antibodies combined with phalloidin staining has revealed antigenically distinct, sorted subsets. Screening of lectin binding capacity indicated wheat germ agglutinin (WGA) as the strongest reactor to amoebocytes. This is further evidenced by WGA inhibition assays that suggest high abundance of N-acetyl-D-glucosamine in amoebocytes. Post-sort phagocytosis assays confirmed the functional differences between amoebocytes and eleocytes, with the former being in favor of bacterial engulfment. This study has proved successful in linking flow cytometry and microscopy analysis and provides further experimental evidence of phenotypic and functional heterogeneity in earthworm coelomocyte subsets.

Item Type: Article
Subjects: Q Science / természettudomány > QR Microbiology / mikrobiológia > QR180 Immunology / immunológia
SWORD Depositor: MTMT SWORD
Depositing User: MTMT SWORD
Date Deposited: 30 Sep 2016 08:06
Last Modified: 30 Sep 2016 08:06
URI: http://real.mtak.hu/id/eprint/40561

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