Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry

Single-cell mass cytometry (SCMC) combines features of traditional flow cytometry (i.e., fluorescence-activated cell sorting) with mass spectrometry, making it possible to measure several parameters at the single-cell level for a complex analysis of biological regulatory mechanisms. In this study, w...

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Autores principales: József Á. Balog, Viktor Honti, Éva Kurucz, Beáta Kari, László G. Puskás, István Andó, Gábor J. Szebeni
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Publicado: Elsevier 2021
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spelling oai:doaj.org-article:2f35895df97a478785d51810ab4becff2021-11-16T04:09:24ZImmunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry1672-022910.1016/j.gpb.2020.06.022https://doaj.org/article/2f35895df97a478785d51810ab4becff2021-04-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S1672022921000553https://doaj.org/toc/1672-0229Single-cell mass cytometry (SCMC) combines features of traditional flow cytometry (i.e., fluorescence-activated cell sorting) with mass spectrometry, making it possible to measure several parameters at the single-cell level for a complex analysis of biological regulatory mechanisms. In this study, we optimized SCMC to analyze hemocytes of the Drosophila innate immune system. We used metal-conjugated antibodies (against cell surface antigens H2, H3, H18, L1, L4, and P1, and intracellular antigens 3A5 and L2) and anti-IgM (against cell surface antigen L6) to detect the levels of antigens, while anti-GFP was used to detect crystal cells in the immune-induced samples. We investigated the antigen expression profile of single cells and hemocyte populations in naive states, in immune-induced states, in tumorous mutants bearing a driver mutation in the Drosophila homologue of Janus kinase (hopTum) and carrying a deficiency of the tumor suppressor gene lethal(3)malignant blood neoplasm-1  [l(3)mbn1], as well as in stem cell maintenance-defective hdcΔ84 mutant larvae. Multidimensional analysis enabled the discrimination of the functionally different major hemocyte subsets for lamellocytes, plasmatocytes, and crystal cells, and delineated the unique immunophenotype of Drosophila mutants. We have identified subpopulations of L2+/P1+ and L2+/L4+/P1+ transitional phenotype cells in the tumorous strains l(3)mbn1 and hopTum, respectively, and a subpopulation of L4+/P1+ cells upon immune induction. Our results demonstrated for the first time that SCMC, combined with multidimensional bioinformatic analysis, represents a versatile and powerful tool to deeply analyze the regulation of cell-mediated immunity of Drosophila.József Á. BalogViktor HontiÉva KuruczBeáta KariLászló G. PuskásIstván AndóGábor J. SzebeniElsevierarticleMass cytometryInnate immunityDrosophilaSingle-cell analysisHemocyteBiology (General)QH301-705.5ENGenomics, Proteomics & Bioinformatics, Vol 19, Iss 2, Pp 243-252 (2021)
institution DOAJ
collection DOAJ
language EN
topic Mass cytometry
Innate immunity
Drosophila
Single-cell analysis
Hemocyte
Biology (General)
QH301-705.5
spellingShingle Mass cytometry
Innate immunity
Drosophila
Single-cell analysis
Hemocyte
Biology (General)
QH301-705.5
József Á. Balog
Viktor Honti
Éva Kurucz
Beáta Kari
László G. Puskás
István Andó
Gábor J. Szebeni
Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry
description Single-cell mass cytometry (SCMC) combines features of traditional flow cytometry (i.e., fluorescence-activated cell sorting) with mass spectrometry, making it possible to measure several parameters at the single-cell level for a complex analysis of biological regulatory mechanisms. In this study, we optimized SCMC to analyze hemocytes of the Drosophila innate immune system. We used metal-conjugated antibodies (against cell surface antigens H2, H3, H18, L1, L4, and P1, and intracellular antigens 3A5 and L2) and anti-IgM (against cell surface antigen L6) to detect the levels of antigens, while anti-GFP was used to detect crystal cells in the immune-induced samples. We investigated the antigen expression profile of single cells and hemocyte populations in naive states, in immune-induced states, in tumorous mutants bearing a driver mutation in the Drosophila homologue of Janus kinase (hopTum) and carrying a deficiency of the tumor suppressor gene lethal(3)malignant blood neoplasm-1  [l(3)mbn1], as well as in stem cell maintenance-defective hdcΔ84 mutant larvae. Multidimensional analysis enabled the discrimination of the functionally different major hemocyte subsets for lamellocytes, plasmatocytes, and crystal cells, and delineated the unique immunophenotype of Drosophila mutants. We have identified subpopulations of L2+/P1+ and L2+/L4+/P1+ transitional phenotype cells in the tumorous strains l(3)mbn1 and hopTum, respectively, and a subpopulation of L4+/P1+ cells upon immune induction. Our results demonstrated for the first time that SCMC, combined with multidimensional bioinformatic analysis, represents a versatile and powerful tool to deeply analyze the regulation of cell-mediated immunity of Drosophila.
format article
author József Á. Balog
Viktor Honti
Éva Kurucz
Beáta Kari
László G. Puskás
István Andó
Gábor J. Szebeni
author_facet József Á. Balog
Viktor Honti
Éva Kurucz
Beáta Kari
László G. Puskás
István Andó
Gábor J. Szebeni
author_sort József Á. Balog
title Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry
title_short Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry
title_full Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry
title_fullStr Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry
title_full_unstemmed Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry
title_sort immunoprofiling of drosophila hemocytes by single-cell mass cytometry
publisher Elsevier
publishDate 2021
url https://doaj.org/article/2f35895df97a478785d51810ab4becff
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