{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T05:50:36Z","timestamp":1776750636355,"version":"3.51.2"},"posted":{"date-parts":[[2023,9,3]]},"group-title":"Immunology","reference-count":59,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,9,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Uncovering the complexity of systems in non-model organisms is critical for understanding arthropod immunology. Prior efforts have mostly focused on Dipteran insects, which only account for a subset of existing arthropod species in nature. Here, we describe immune cells or hemocytes from the clinically relevant tick\n                  <jats:italic>Ixodes scapularis<\/jats:italic>\n                  using bulk and single cell RNA sequencing combined with depletion via clodronate liposomes, RNA interference, Clustered Regularly Interspaced Short Palindromic Repeats activation (CRISPRa) and RNA-fluorescence\n                  <jats:italic>in situ<\/jats:italic>\n                  hybridization (FISH). We observe molecular alterations in hemocytes upon tick infestation of mammals and infection with either the Lyme disease spirochete\n                  <jats:italic>Borrelia burgdorferi<\/jats:italic>\n                  or the rickettsial agent\n                  <jats:italic>Anaplasma phagocytophilum<\/jats:italic>\n                  . We predict distinct hemocyte lineages and reveal clusters exhibiting defined signatures for immunity, metabolism, and proliferation during hematophagy. Furthermore, we perform a mechanistic characterization of two\n                  <jats:italic>I. scapularis<\/jats:italic>\n                  hemocyte markers:\n                  <jats:italic>hemocytin<\/jats:italic>\n                  and\n                  <jats:italic>astakine<\/jats:italic>\n                  . Depletion of phagocytic hemocytes affects\n                  <jats:italic>hemocytin<\/jats:italic>\n                  and\n                  <jats:italic>astakine<\/jats:italic>\n                  levels, which impacts blood feeding and molting behavior of ticks. Hemocytin specifically affects the c-Jun N-terminal kinase (JNK) signaling pathway, whereas astakine alters hemocyte proliferation in\n                  <jats:italic>I. scapularis<\/jats:italic>\n                  . Altogether, we uncover the heterogeneity and pleiotropic roles of hemocytes in ticks and provide a valuable resource for comparative biology in arthropods.\n                <\/jats:p>","DOI":"10.1101\/2023.08.31.555785","type":"posted-content","created":{"date-parts":[[2023,9,3]],"date-time":"2023-09-03T14:30:11Z","timestamp":1693751411000},"source":"Crossref","is-referenced-by-count":4,"title":["Tick hemocytes have pleiotropic roles in microbial infection and arthropod fitness"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1059-3578","authenticated-orcid":false,"given":"Agustin","family":"Rolandelli","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanna J.","family":"Laukaitis-Yousey","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haikel N.","family":"Bogale","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nisha","family":"Singh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sourabh","family":"Samaddar","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anya J.","family":"O\u2019Neal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Camila R.","family":"Ferraz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew","family":"Butnaru","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0172-8090","authenticated-orcid":false,"given":"Enzo","family":"Mameli","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2536-0267","authenticated-orcid":false,"given":"Baolong","family":"Xia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. Tays","family":"Mendes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"L. Rainer","family":"Butler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liron","family":"Marnin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francy E.","family":"Cabrera Paz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luisa M.","family":"Valencia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vipin S.","family":"Rana","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ciaran","family":"Skerry","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Utpal","family":"Pal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9639-7708","authenticated-orcid":false,"given":"Stephanie E.","family":"Mohr","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7542-472X","authenticated-orcid":false,"given":"Norbert","family":"Perrimon","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Serre","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8740-2667","authenticated-orcid":false,"given":"Joao H.F.","family":"Pedra","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023090706500464000_2023.08.31.555785v1.1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-ento-010715-023924"},{"key":"2023090706500464000_2023.08.31.555785v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2018.09.003"},{"key":"2023090706500464000_2023.08.31.555785v1.3","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1016\/j.pt.2020.07.010","article-title":"Immunometabolism in arthropod vectors: redefining interspecies relationships","volume":"36","year":"2020","journal-title":"Trends in Parasitology"},{"key":"2023090706500464000_2023.08.31.555785v1.4","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msv093"},{"key":"2023090706500464000_2023.08.31.555785v1.5","doi-asserted-by":"crossref","unstructured":"J. Bechsgaard et al., Comparative genomic study of arachnid immune systems indicates loss of \u03b2GRPs and the IMD pathway. J Evol Biol 29, 19 (2015).","DOI":"10.1111\/jeb.12780"},{"key":"2023090706500464000_2023.08.31.555785v1.6","doi-asserted-by":"crossref","unstructured":"D. K. Shaw et al., Infection-derived lipids elicit an immune deficiency circuit in arthropods. Nature Communications 8, 14401 (2017).","DOI":"10.1038\/ncomms14401"},{"key":"2023090706500464000_2023.08.31.555785v1.7","doi-asserted-by":"crossref","first-page":"e2208673120","DOI":"10.1073\/pnas.2208673120","article-title":"Croquemort elicits activation of the immune deficiency pathway in ticks","volume":"120","year":"2023","journal-title":"Proc Natl Acad Sci USA"},{"key":"2023090706500464000_2023.08.31.555785v1.8","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1808905116"},{"key":"2023090706500464000_2023.08.31.555785v1.9","doi-asserted-by":"crossref","unstructured":"V. S. Rana et al., Dome1-JAK-STAT signaling between parasite and host integrates vector immunity and development. Science 379, eabl3837 (2023).","DOI":"10.1126\/science.abl3837"},{"key":"2023090706500464000_2023.08.31.555785v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2016.06.001"},{"key":"2023090706500464000_2023.08.31.555785v1.11","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.21.9465"},{"key":"2023090706500464000_2023.08.31.555785v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.dci.2013.05.014"},{"key":"2023090706500464000_2023.08.31.555785v1.13","doi-asserted-by":"publisher","DOI":"10.1101\/gad.12.20.3252"},{"key":"2023090706500464000_2023.08.31.555785v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.idc.2015.02.007"},{"key":"2023090706500464000_2023.08.31.555785v1.15","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2017.12.006"},{"key":"2023090706500464000_2023.08.31.555785v1.16","doi-asserted-by":"crossref","first-page":"587","DOI":"10.1038\/s41579-020-0400-5","article-title":"Interactions between Borrelia burgdorferi and ticks","volume":"18","year":"2020","journal-title":"Nature Reviews Microbiology"},{"key":"2023090706500464000_2023.08.31.555785v1.17","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1146\/annurev-ento-020117-043027","article-title":"Phylogeography of ticks (Acari: Ixodida)","volume":"64","year":"2019","journal-title":"Annual Review of Entomology"},{"key":"2023090706500464000_2023.08.31.555785v1.18","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1038\/s41588-022-01275-w","article-title":"A high-quality Ixodes scapularis genome advances tick science","volume":"55","year":"2023","journal-title":"Nat Genet"},{"key":"2023090706500464000_2023.08.31.555785v1.19","doi-asserted-by":"publisher","DOI":"10.1016\/j.dci.2015.12.006"},{"key":"2023090706500464000_2023.08.31.555785v1.20","doi-asserted-by":"publisher","DOI":"10.2307\/3277874"},{"key":"2023090706500464000_2023.08.31.555785v1.21","doi-asserted-by":"publisher","DOI":"10.1093\/jmedent\/7.3.277"},{"key":"2023090706500464000_2023.08.31.555785v1.22","doi-asserted-by":"crossref","unstructured":"J. M. Starck et al., Morphological responses to feeding in ticks (Ixodes ricinus). Zoological Lett 4, 20 (2018).","DOI":"10.1186\/s40851-018-0104-0"},{"key":"2023090706500464000_2023.08.31.555785v1.23","doi-asserted-by":"crossref","unstructured":"I. Soderhall , E. Fasterius , C. Ekblom , K. Soderhall , Characterization of hemocytes and hematopoietic cells of a freshwater crayfish based on single-cell transcriptome analysis. iScience 25, 104850 (2022).","DOI":"10.1016\/j.isci.2022.104850"},{"key":"2023090706500464000_2023.08.31.555785v1.24","doi-asserted-by":"publisher","DOI":"10.1016\/j.ydbio.2003.06.001"},{"key":"2023090706500464000_2023.08.31.555785v1.25","doi-asserted-by":"publisher","DOI":"10.1016\/j.dci.2007.03.012"},{"key":"2023090706500464000_2023.08.31.555785v1.26","doi-asserted-by":"crossref","unstructured":"P. Sukonthamarn , Z. Nanakorn , W. Junprung , P. Supungul , A. Tassanakajon , Role of hemocytin from Litopenaeus vannamei in immune response against microsporidian, Enterocytozoon hepatopenaei . Fish Shellfish Immunol 136, 108710 (2023).","DOI":"10.1016\/j.fsi.2023.108710"},{"key":"2023090706500464000_2023.08.31.555785v1.27","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.174.10.6153"},{"key":"2023090706500464000_2023.08.31.555785v1.28","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.dci.2015.10.025","article-title":"A cytokine-like factor astakine accelerates the hemocyte production in Pacific oyster Crassostrea gigas","volume":"55","year":"2016","journal-title":"Developmental and Comparative Immunology"},{"key":"2023090706500464000_2023.08.31.555785v1.29","doi-asserted-by":"crossref","unstructured":"J. L. Benton et al., Adult neurogenesis in crayfish: Identity and regulation of neural progenitors produced by the immune system. iScience 25, 103993 (2022).","DOI":"10.1016\/j.isci.2022.103993"},{"key":"2023090706500464000_2023.08.31.555785v1.30","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/j.dci.2017.11.005","article-title":"Astakines in arthropods-phylogeny and gene structure","volume":"81","year":"2018","journal-title":"Developmental and Comparative Immunology"},{"key":"2023090706500464000_2023.08.31.555785v1.31","doi-asserted-by":"crossref","unstructured":"A. Adegoke , J. M. C. Ribeiro , S. Brown , R. C. Smith , S. Karim , Rickettsia parkeri hijacks tick hemocytes to manipulate cellular and humoral transcriptional responses. Frontiers in Immunology 14, 1094326 (2023).","DOI":"10.3389\/fimmu.2023.1094326"},{"key":"2023090706500464000_2023.08.31.555785v1.32","doi-asserted-by":"crossref","unstructured":"A. Sharma et al., Cas9-mediated gene editing in the black-legged tick, Ixodes scapularis, by embryo injection and ReMOT Control. iScience 25, 103781 (2022).","DOI":"10.1016\/j.isci.2022.103781"},{"key":"2023090706500464000_2023.08.31.555785v1.33","doi-asserted-by":"publisher","DOI":"10.1016\/j.dci.2007.11.002"},{"key":"2023090706500464000_2023.08.31.555785v1.34","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-010-0364-0"},{"key":"2023090706500464000_2023.08.31.555785v1.35","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2013.06.044"},{"key":"2023090706500464000_2023.08.31.555785v1.36","doi-asserted-by":"crossref","unstructured":"J. Ramesh Kumar , J. P. Smith , H. Kwon , R. C. Smith , Use of clodronate liposomes to deplete phagocytic immune cells in Drosophila melanogaster and Aedes aegypti . Front Cell Dev Biol 9, 627976 (2021).","DOI":"10.3389\/fcell.2021.627976"},{"key":"2023090706500464000_2023.08.31.555785v1.37","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1900147116"},{"key":"2023090706500464000_2023.08.31.555785v1.38","doi-asserted-by":"crossref","unstructured":"H. Kwon , M. Mohammed , O. Franzen , J. Ankarklev , R. C. Smith , Single-cell analysis of mosquito hemocytes identifies signatures of immune cell subtypes and cell differentiation. eLife 10, (2021).","DOI":"10.7554\/eLife.66192"},{"key":"2023090706500464000_2023.08.31.555785v1.39","doi-asserted-by":"publisher","DOI":"10.1126\/science.abc0322"},{"key":"2023090706500464000_2023.08.31.555785v1.40","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1803062115"},{"key":"2023090706500464000_2023.08.31.555785v1.41","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1007\/s13355-019-00631-3","article-title":"Steroid hormones in Acari, their functions and synthesis","volume":"54","year":"2019","journal-title":"Applied Entomology and Zoology"},{"key":"2023090706500464000_2023.08.31.555785v1.42","doi-asserted-by":"publisher","DOI":"10.1038\/embor.2011.177"},{"key":"2023090706500464000_2023.08.31.555785v1.43","doi-asserted-by":"crossref","unstructured":"A. Mase , J. Augsburger , K. Br\u00fcckner , Macrophages and their organ locations shape each other in development and homeostasis \u2013 a Drosophila perspective. Frontiers in Cell and Developmental Biology 9, 630272 (2021).","DOI":"10.3389\/fcell.2021.630272"},{"key":"2023090706500464000_2023.08.31.555785v1.44","doi-asserted-by":"crossref","unstructured":"D. Stanley , E. Haas , Y. Kim , Beyond cellular immunity: on the biological significance of insect hemocytes. Cells 12, 599 (2023).","DOI":"10.3390\/cells12040599"},{"key":"2023090706500464000_2023.08.31.555785v1.45","doi-asserted-by":"crossref","first-page":"3696","DOI":"10.1038\/s41467-021-23900-8","article-title":"Tick extracellular vesicles enable arthropod feeding and promote distinct outcomes of bacterial infection","volume":"12","year":"2021","journal-title":"Nature Communications"},{"key":"2023090706500464000_2023.08.31.555785v1.46","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.68.3.1125-1133.2000"},{"key":"2023090706500464000_2023.08.31.555785v1.47","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.1.446-455.2001"},{"key":"2023090706500464000_2023.08.31.555785v1.48","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.3317"},{"key":"2023090706500464000_2023.08.31.555785v1.49","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btp616"},{"key":"2023090706500464000_2023.08.31.555785v1.50","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.54818"},{"key":"2023090706500464000_2023.08.31.555785v1.51","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2023438118"},{"key":"2023090706500464000_2023.08.31.555785v1.52","doi-asserted-by":"crossref","unstructured":"A. T. Lun , K. Bach , J. C. Marioni , Pooling across cells to normalize single-cell RNA sequencing data with many zero counts. Genome Biology 17, 75 (2016).","DOI":"10.1186\/s13059-016-0947-7"},{"key":"2023090706500464000_2023.08.31.555785v1.53","first-page":"2122","article-title":"A step-by-step workflow for low-level analysis of single-cell RNA-seq data with Bioconductor","volume":"5","year":"2016","journal-title":"F1000Res"},{"key":"2023090706500464000_2023.08.31.555785v1.54","doi-asserted-by":"crossref","unstructured":"K. Street et al., Slingshot: cell lineage and pseudotime inference for single-cell transcriptomics. BMC Genomics 19, 477 (2018).","DOI":"10.1186\/s12864-018-4772-0"},{"key":"2023090706500464000_2023.08.31.555785v1.55","doi-asserted-by":"crossref","unstructured":"G. Finak et al., MAST: a flexible statistical framework for assessing transcriptional changes and characterizing heterogeneity in single-cell RNA sequencing data. Genome Biology 16, 278 (2015).","DOI":"10.1186\/s13059-015-0844-5"},{"key":"2023090706500464000_2023.08.31.555785v1.56","doi-asserted-by":"crossref","first-page":"D898","DOI":"10.1093\/nar\/gkab929","article-title":"VEuPathDB: the eukaryotic pathogen, vector and host bioinformatics resource center","volume":"50","year":"2022","journal-title":"Nucleic Acids Research"},{"key":"2023090706500464000_2023.08.31.555785v1.57","doi-asserted-by":"crossref","unstructured":"L. S. Gramates et al., FlyBase: a guided tour of highlighted features. Genetics 220, (2022).","DOI":"10.1093\/genetics\/iyac035"},{"key":"2023090706500464000_2023.08.31.555785v1.58","doi-asserted-by":"publisher","DOI":"10.1093\/NAR\/GKAC993"},{"key":"2023090706500464000_2023.08.31.555785v1.59","doi-asserted-by":"publisher","DOI":"10.1093\/NAR\/GKAC1052"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2023.08.31.555785","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T05:09:56Z","timestamp":1768453796000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2023.08.31.555785"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,3]]},"references-count":59,"URL":"https:\/\/doi.org\/10.1101\/2023.08.31.555785","relation":{},"subject":[],"published":{"date-parts":[[2023,9,3]]},"subtype":"preprint"}}