Immunoprofiling of Drosophila Hemocytes by Single-cell Mass Cytometry  

作　　者：Jozsef A.Balog Viktor Honti Eva Kurucz Beata Kari Laszlo G.Puskas Istvan Ando Gabor J.Szebeni 

机构地区：[1]Laboratory of Functional Genomics,Institute of Genetics,Biological Research Centre,Szeged H-6726,Hungary [2]University of Szeged,Ph.D.School in Biology,Szeged H-6726,Hungary [3]Immunology Unit,Institute of Genetics,Biological Research Centre,Szeged H-6726,Hungary [4]Department of Physiology,Anatomy and Neuroscience,Faculty of Science and Informatics,University of Szeged,Szeged H-6726,Hungary

出　　处：《Genomics, Proteomics & Bioinformatics》2021年第2期243-252,共10页基因组蛋白质组与生物信息学报（英文版）

基　　金：the National Research,Development and Innovation Office,Hungary(Grant Nos.GINOP-2.3.2-15-2016-00001,GINOP-2.3.2-15-2016-00030 to LGP,GINOP-2.3.2-15-2016-00035 to E´K,NKFI NN118207 and NKFI K120142 to IA,NKFI 120140 to EK,and OTKA K-131484 to VH);Gabor J.Szebeni was supported by the New National Excellence Program of the Ministry for Innovation and Technology,Hungary(Grant No.UNKP-19-4-SZTE-36)and by the Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences(Grant No.BO/00139/17/8)。

摘　　要：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 hdcD84 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.

关 键 词：Mass cytometry Innate immunity DROSOPHILA Single-cell analysis HEMOCYTE 

分 类 号：Q939.91[生物学—微生物学]