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

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.

journal_name

Dev Comp Immunol

journal_title

Developmental and comparative immunology

authors

Engelmann P,Hayashi Y,Bodó K,Ernszt D,Somogyi I,Steib A,Orbán J,Pollák E,Nyitrai M,Németh P,Molnár L

doi

10.1016/j.dci.2016.06.017

subject

Has Abstract

pub_date

2016-12-01 00:00:00

pages

41-52

eissn

0145-305X

issn

1879-0089

pii

S0145-305X(16)30207-5

journal_volume

65

pub_type

杂志文章

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