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

Péter Engelmann; Yuya Hayashi; Kornélia Bodo; David Ernszt; Ildiko Somogyi; Anita Steib; Jozsef Orban; Edit  Pollak; Miklos Nyitrai; Péter Németh; Laszlo Molnar

doi:10.1016/j.dci.2016.06.017

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

Péter Engelmann, Yuya Hayashi, Kornélia Bodo, David Ernszt, Ildiko Somogyi, Anita Steib, Jozsef Orban, Edit Pollak, Miklos Nyitrai, Péter Németh, Laszlo Molnar

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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.

Original language	English
Journal	Developmental & Comparative Immunology
Volume	65
Pages (from-to)	41-52
Number of pages	12
ISSN	0145-305X
DOIs	https://doi.org/10.1016/j.dci.2016.06.017
Publication status	Published - 24 Jun 2016

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Engelmann, P., Hayashi, Y., Bodo, K., Ernszt, D., Somogyi, I., Steib, A., Orban, J., Pollak, E., Nyitrai, M., Németh, P., & Molnar, L. (2016). Phenotypic and functional characterization of earthworm coelomocyte subsets: Linking light scatter-based cell typing and imaging of the sorted populations. Developmental & Comparative Immunology, 65, 41-52. https://doi.org/10.1016/j.dci.2016.06.017

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title = "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.",

author = "P{\'e}ter Engelmann and Yuya Hayashi and Korn{\'e}lia Bodo and David Ernszt and Ildiko Somogyi and Anita Steib and Jozsef Orban and Edit Pollak and Miklos Nyitrai and P{\'e}ter N{\'e}meth and Laszlo Molnar",

year = "2016",

month = jun,

day = "24",

doi = "10.1016/j.dci.2016.06.017",

language = "English",

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journal = "Developmental & Comparative Immunology",

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Engelmann, P, Hayashi, Y, Bodo, K, Ernszt, D, Somogyi, I, Steib, A, Orban, J, Pollak, E, Nyitrai, M, Németh, P & Molnar, L 2016, 'Phenotypic and functional characterization of earthworm coelomocyte subsets: Linking light scatter-based cell typing and imaging of the sorted populations', Developmental & Comparative Immunology, vol. 65, pp. 41-52. https://doi.org/10.1016/j.dci.2016.06.017

Phenotypic and functional characterization of earthworm coelomocyte subsets: Linking light scatter-based cell typing and imaging of the sorted populations. / Engelmann, Péter; Hayashi, Yuya; Bodo, Kornélia et al.
In: Developmental & Comparative Immunology, Vol. 65, 24.06.2016, p. 41-52.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Phenotypic and functional characterization of earthworm coelomocyte subsets

T2 - Linking light scatter-based cell typing and imaging of the sorted populations

AU - Engelmann, Péter

AU - Hayashi, Yuya

AU - Bodo, Kornélia

AU - Ernszt, David

AU - Somogyi, Ildiko

AU - Steib, Anita

AU - Orban, Jozsef

AU - Pollak, Edit

AU - Nyitrai, Miklos

AU - Németh, Péter

AU - Molnar, Laszlo

PY - 2016/6/24

Y1 - 2016/6/24

N2 - 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.

AB - 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.

U2 - 10.1016/j.dci.2016.06.017

DO - 10.1016/j.dci.2016.06.017

M3 - Journal article

C2 - 27349970

SN - 0145-305X

VL - 65

SP - 41

EP - 52

JO - Developmental & Comparative Immunology

JF - Developmental & Comparative Immunology

ER -

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

Abstract

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