Anaplasma phagocytophilum is an emerging pathogen that causes human granulocytic anaplasmosis. Infection with this zoonotic pathogen affects cell function in both vertebrate host and the tick vector, Ixodes scapularis. Global tissue-specific response and apoptosis signaling pathways were characterized in I. scapularis nymphs and adult female midguts and salivary glands infected with A. phagocytophilum using a systems biology approach combining transcriptomics and proteomics. Apoptosis was selected for pathway-focused analysis due to its role in bacterial infection of tick cells. The results showed tissue-specific differences in tick response to infection and revealed differentiated regulation of apoptosis pathways. The impact of bacterial infection was more pronounced in tick nymphs and midguts than in salivary glands, probably reflecting bacterial developmental cycle. All apoptosis pathways described in other organisms were identified in I. scapularis, except for the absence of the Perforin ortholog. Functional characterization using RNA interference showed that Porin knockdown significantly increases tick colonization by A. phagocytophilum. Infection with A. phagocytophilum produced complex tissue-specific alterations in transcript and protein levels. In tick nymphs, the results suggested a possible effect of bacterial infection on the inhibition of tick immune response. In tick midguts, the results suggested that A. phagocytophilum infection inhibited cell apoptosis to facilitate and establish infection through up-regulation of the JAK/STAT pathway. Bacterial infection inhibited the intrinsic apoptosis pathway in tick salivary glands by down-regulating Porin expression that resulted in the inhibition of Cytochrome c release as the anti-apoptotic mechanism to facilitate bacterial infection. However, tick salivary glands may promote apoptosis to limit bacterial infection through induction of the extrinsic apoptosis pathway. These dynamic changes in response to A. phagocytophilum in I. scapularis tissue-specific transcriptome and proteome demonstrated the complexity of the tick response to infection and will contribute to characterize gene regulation in ticks.

Centro Nacional de Investigaciones Cardiovasculares Madrid Spain

Department of Veterinary Pathobiology Center for Veterinary Health Sciences Oklahoma State University Stillwater Oklahoma United States of America

Institute of Parasitology Biology Centre Academy of Sciences of the Czech Republic České Budějovice The Czech Republic

SaBio Instituto de Investigación en Recursos Cinegéticos IREC CSIC UCLM JCCM Ciudad Real Spain

SaBio Instituto de Investigación en Recursos Cinegéticos IREC CSIC UCLM JCCM Ciudad Real Spain; Center for Infection and Immunity of Lille Université Lille Nord de France Institut Pasteur de Lille Lille France

SaBio Instituto de Investigación en Recursos Cinegéticos IREC CSIC UCLM JCCM Ciudad Real Spain; Department of Veterinary Pathobiology Center for Veterinary Health Sciences Oklahoma State University Stillwater Oklahoma United States of America

Zobrazit více v PubMed

de la Fuente J, Estrada-Peña A, Venzal JM, Kocan KM, Sonenshine DE (2008) Overview: Ticks as vectors of pathogens that cause disease in humans and animals. Front Biosci 13: 6938–6946. PubMed

Reichard MV, Manzano-Roman R, Kocan KM, Blouin EF, de la Fuente J, et al. (2009) Inoculation of white-tailed deer (Odocoileus virginianus) with Ap-V1 or NY-18 strains of Anaplasma phagocytophilum and microscopic demonstration of Ap-V1 in Ixodes scapularis adults that acquired infection from deer as nymphs. Vector Borne Zoonotic Dis 9: 565–568. 10.1089/vbz.2008.0106 PubMed DOI

Sukumaran B, Narasimham S, Anderson JF, DePonte K, Marcantonio K, et al. (2006) An Ixodes scapularis protein required for survival of Anaplasma phagocytophilum in tick salivary glands. J Exp Med 6: 1507–1517. PubMed PMC

Dumler JS, Barbet AC, Bekker CPJ, Dasch GA, Palmer GH, et al. (2001) Reorganization of the genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions subjective synonyms of Ehrlichia phagocytophila . Int J Syst Evol Microbiol 51: 2145–2165. PubMed

Rikihisa Y (2011) Molecular pathogensis of Anaplasma phagocytophilum . Current Microbiol Rev 24: 469–489. 10.1128/CMR.00064-10 PubMed DOI PMC

Severo MS, Choy A, Stephens KD, Sakhon OS, Chen G, et al. (2013) The E3 ubiquitin ligase XIAP restricts Anaplasma phagocytophilum colonization of Ixodes scapularis ticks. J Infect Dis 208: 1830–1840. 10.1093/infdis/jit380 PubMed DOI PMC

Stuen S (2010) Anaplasma phagocytophilum—the most widespread tick-borne infection in animals in Europe. Vet Res Commun 31: 79–84. PubMed

Stuen S, Granquist EG, Silaghi C (2013) Anaplasma phagocytophilum—a widespread multi-host pathogen with highly adaptive strategies. Front Cell Infect Microbiol 3: 31 10.3389/fcimb.2013.00031 PubMed DOI PMC

Dahlgren FS, Mandel EJ, Krebs JW, Massung RF, McQuiston JH (2011) Increasing incidence of Ehrlichia chaffeensis and Anaplasma phagocytophilum in the United States, 2000–2007. Am J Trop Med Hyg 85: 124–131. 10.4269/ajtmh.2011.10-0613 PubMed DOI PMC

Carlyon JA, Fikrig E (2003) Invasion and survival strategies of Anaplasma phagocytophilum . Cell Microbiol 5: 743–754. PubMed

Lee HC, Goodman JL (2006) Anaplasma phagocytophilum causes global induction of antiapoptosis in human neutrophils. Genomics 88: 496–503. PubMed

Lee HC, Kioi M, Han J, Puri RK, Goodman JL (2008) Anaplasma phagocytophilum-induced gene expression in both human neutrophils and HL-60 cells. Genomics 92: 144–151. 10.1016/j.ygeno.2008.05.005 PubMed DOI

Galindo RC, de la Fuente J (2012) Transcriptomics data integration reveals Jak-STAT pathway as a common pathway affected by pathogenic intracellular bacteria in natural reservoir hosts. J Proteomics Bioinformatics 5: 108–115.

Woldehiwet Z, Yavari C (2014) Anaplasma phagocytophilum up-regulates some anti-apoptotic genes in neutrophils and pro-inflammatory genes in mononuclear cells of sheep. J Comp Pathol 150: 351–356. 10.1016/j.jcpa.2014.01.005 PubMed DOI

Hajdušek O, Šíma R, Ayllón N, Jalovecká M, Perner J, et al. (2013) Interaction of the tick immune system with transmitted pathogens. Front Cell Infect Microbiol 3: 26 10.3389/fcimb.2013.00026 PubMed DOI PMC

Ayllón N, Villar M, Busby AT, Kocan KM, Blouin EF, et al. (2013) Anaplasma phagocytophilum inhibits apoptosis and promotes cytoskeleton rearrangement for infection of tick cells. Infect Immun 81: 2415–2425. 10.1128/IAI.00194-13 PubMed DOI PMC

Sunyakumthorn P, Petchampai N, Grasperge BJ, Kearney MT, Sonenshine DE, et al. (2013) Gene expression of tissue-specific molecules in ex vivo Dermacentor variabilis (Acari: Ixodidae) during rickettsial exposure. J Med Entomol 50: 1089–1096. PubMed PMC

Brown JB, Boley N, Eisman R, May GE, Stoiber MH, et al. (2014) Diversity and dynamics of the Drosophila transcriptome. Nature 512: 393–399. PubMed PMC

de la Fuente J, Kocan KM, Blouin EF, Zivkovic Z, Naranjo V, et al. (2010) Functional genomics and evolution of tick-Anaplasma interactions and vaccine development. Vet Parasitol 167: 175–186. 10.1016/j.vetpar.2009.09.019 PubMed DOI

de la Fuente J, Merino O (2013) Vaccinomics, the new road to tick vaccines. Vaccine 31: 5923–5929. PubMed

Ekblom R, Galindo J (2011) Applications of next generation sequencing in molecular ecology of non-model organisms. Heredity (Edinb) 107: 1–15. PubMed PMC

Mochida K, Shinozaki K (2011) Advances in omics and bioinformatics tools for systems analyses of plant functions. Plant Cell Physiol 52: 2017–2038. 10.1093/pcp/pcr153 PubMed DOI PMC

Villar M, Popara M, Ayllón N, Fernández de Mera IG, Mateos-Hernández L, et al. (2014) A systems biology approach to the characterization of stress response in Dermacentor reticulatus tick unfed larvae. PLoS ONE 9: e89564 10.1371/journal.pone.0089564 PubMed DOI PMC

Naranjo N, Ayllón N, Pérez de la Lastra JM, Galindo RC, Kocan KM, et al. (2013) Reciprocal regulation of NF-kB (Relish) and Subolesin in the tick vector, Ixodes scapularis . PLoS ONE 8: e65915 10.1371/journal.pone.0065915 PubMed DOI PMC

Genomic Resources Development Consortium, Contreras M, de la Fuente J, Estrada-Peña A, Grubhoffer L, Tobes R (2014) Transcriptome sequence divergence between Lyme disease tick vectors, Ixodes scapularis and Ixodes ricinus. Genomic Resources Notes accepted 1 April 2014–31 May 2014. Mol Ecol Resour 14: 1095 10.1111/1755-0998.12298 PubMed DOI

Mastronunzio JE, Kurscheid S, Fikrig E (2012) Postgenomic analyses reveal development of infectious Anaplasma phagocytophilum during transmission from ticks to mice. J Bacteriol 194: 2238–2247. 10.1128/JB.06791-11 PubMed DOI PMC

Sinclair SH, Rennoll-Bankert KE, Dumler JS (2014) Effector bottleneck: microbial reprogramming of parasitized host cell transcription by epigenetic remodeling of chromatin structure. Front Genet 5: 274 10.3389/fgene.2014.00274 PubMed DOI PMC

Geraci NS, Johnston JS, Robinson JP, Wikel SK, Hill CA (2007) Variation in genome size of argasid and ixodid ticks. Insect Biochem Mol Biol 37: 399–408. PubMed

de Sousa Abreu R, Penalva LO, Marcotte EM, Vogel C (2009) Global signatures of protein and mRNA expression levels. Mol Biosyst 5: 1512–1526. 10.1039/b908315d PubMed DOI PMC

Hirsch CL, Bonham K (2004) Histone deacetylase inhibitors regulate p21WAF1 gene expression at the post-transcriptional level in HepG2 cells. FEBS Lett 570: 37–40. PubMed

Lin M, den Dulk-Ras A, Hooykaas PJ, Rikihisa Y (2007) Anaplasma phagocytophilum AnkA secreted by type IV secretion system is tyrosine phosphorylated by Abl-1 to facilitate infection. Cell Microbiol 9: 2644–2657. PubMed

Elmore S (2007) Apoptosis: A review of programmed cell death. Toxicol Pathol 35: 496–516. PubMed PMC

de la Fuente J, Kocan KM, Almazán C, Blouin EF (2007) RNA interference for the study and genetic manipulation of ticks. Trends Parasitol 23: 427–433. PubMed

Lindsay J, Esposti MD, Gilmore AP (2011) Bcl-2 proteins and mitochondria—specificity in membrane targeting for death. Biochim Biophys Acta 1813: 532–539. 10.1016/j.bbamcr.2010.10.017 PubMed DOI

Sul HS, Wang D (1998) Nutritional and hormonal regulation of enzymes in fat synthesis: studies of fatty acid synthase and mitochondrial glycerol-3-phosphate acyltransferase gene transcription. Annu Rev Nutr. 1998;18:331–51. PubMed

Bandyopadhyay S, Zhan R, Wang Y, Pai SK, Hirota S, et al. (2006) Mechanism of apoptosis induced by the inhibition of fatty acid synthase in breast cancer cells. Cancer Res 66: 5934–5940. PubMed

Zhang JH, Zhang Y, Herman B (2003) Caspases, apoptosis and aging. Ageing Res Reviews 2: 357–366. PubMed

Zecchin KG, Rossato FA, Raposo HF, Melo DR, Alberici LC, et al. (2011) Inhibition of fatty acid synthase in melanoma cells activates the intrinsic pathway of apoptosis. Lab Invest 91: 232–240. 10.1038/labinvest.2010.157 PubMed DOI

Rossato FA, Zecchin KG, La Guardia PG, Ortega RM, Alberici LC, et al. (2014) Fatty acid synthase inhibitors induce apoptosis in non-tumorigenic melan-a cells associated with inhibition of mitochondrial respiration. PLoS ONE 9: e101060 10.1371/journal.pone.0101060 PubMed DOI PMC

Zuo FY, Li SY, An P, Yu B, Cai HY (2004) The construction of yeast two-hybrid method in the protein-interactions and its significance in hepatic metastasis of colorectal carcinoma. Zhonghua Wai Ke Za Zhi 42: 672–674. PubMed

Vandhana S, Coral K, Jayanthi U, Deepa PR, Krishnakumar S (2013) Biochemical changes accompanying apoptotic cell death in retinoblastoma cancer cells treated with lipogenic enzyme inhibitors. Biochim Biophys Acta 1831: 1458–1466. 10.1016/j.bbalip.2013.06.005 PubMed DOI

Kocan KM, Busby AT, Allison RW, Breshears MA, Coburn L, et al. (2012) Sheep experimentally-infected with a human isolate of Anaplasma phagocytophilum serve as a host for infection of Ixodes scapularis . Ticks Tick-Borne Dis 3: 147–153. 10.1016/j.ttbdis.2012.01.004 PubMed DOI

Trapnell C, Pachter L, Salzberg SL (2009) TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 25: 1105–1111. 10.1093/bioinformatics/btp120 PubMed DOI PMC

Langmead B, Trapnell C, Pop M, Salzberg SL (2009) Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol 10: R25 10.1186/gb-2009-10-3-r25 PubMed DOI PMC

Wang L, Feng Z, Wang X, Wang X, Zhang X (2010) DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics 26:136–138. 10.1093/bioinformatics/btp612 PubMed DOI

Wisniewski J, Zougman A, Nagaraj N, Mann M (2009) Universal sample preparation method for proteome analysis. Nat Meth 6: 359–362. PubMed

Martínez-Bartolomé S, Navarro P, Martín-Maroto F, López-Ferrer D, Ramos-Fernández A, et al. (2008) Properties of average score distributions of SEQUEST: the probability ratio method. Mol Cell Proteomics 7: 1135–1145. 10.1074/mcp.M700239-MCP200 PubMed DOI

Navarro P, Vázquez J (2009) A refined method to calculate false discovery rates for peptide identification using decoy databases. J Proteome Res 8: 1792–1796. 10.1021/pr800362h PubMed DOI

Bonzon-Kulichenko E, Martínez-Martínez S, Trevisan-Herraz M, Navarro P, Redondo JM, et al. (2011) Quantitative in-depth analysis of the dynamic secretome of activated Jurkat T-cells. J Proteomics 75: 561–571. 10.1016/j.jprot.2011.08.022 PubMed DOI

Jorge I, Navarro P, Martinez-Acedo P, Nunez E, Serrano H, et al. (2009) Statistical model to analyze quantitative proteomics data obtained by 18O/16O labeling and linear ion trap mass spectrometry: application to the study of vascular endothelial growth factor-induced angiogenesis in endothelial cells. Mol Cell Proteomics 8: 1130–1149. 10.1074/mcp.M800260-MCP200 PubMed DOI PMC

Navarro P, Trevisan-Herraz M, Bonzon-Kulichenko E, Núñez E, Martínez-Acedo P, et al. (2014) General Statistical framework for quantitative proteomics by stable isotope labeling. J Proteome Res 13: 1234–1247. 10.1021/pr4006958 PubMed DOI

Bhatia VN, Perlman DH, Costello CE, McComb ME (2009) Software tool for researching annotations of proteins: Open-source protein annotation software with data visualization. Anal Chem 81: 9819–9823. 10.1021/ac901335x PubMed DOI PMC

Kocan KM, Blouin E, de la Fuente J (2011) RNA interference in ticks. JoVE 47: e2474 http://www.jove.com/details.stp?id=2474 PubMed PMC

Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32: 1792–1797. PubMed PMC

Castresana J (2000) Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. Mol Biol Evol 17: 540–552. PubMed

Anisimova M, Gascuel O (2006) Approximate likelihood ratio test for branches: a fast, accurate and powerful alternative. Syst Biol 55: 539–552. PubMed

Guindon S, Gascuel O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol 52: 696–704. PubMed

Chevenet F, Brun C, Banuls AL, Jacq B, Chisten R (2006) TreeDyn: towards dynamic graphics and annotations for analyses of trees. BMC Bioinf 10: 439. PubMed PMC

Experimental Infection of Mice and Ticks with the Human Isolate of Anaplasma phagocytophilum NY-18

Tick Immune System: What Is Known, the Interconnections, the Gaps, and the Challenges

Enlisting the Ixodes scapularis Embryonic ISE6 Cell Line to Investigate the Neuronal Basis of Tick-Pathogen Interactions

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1

Comparative proteomics of the vector Dermacentor reticulatus revealed differentially regulated proteins associated with pathogen transmission in response to laboratory infection with Rickettsia slovaca

Counterattacking the tick bite: towards a rational design of anti-tick vaccines targeting pathogen transmission

Functional Evolution of Subolesin/Akirin

Tick galactosyltransferases are involved in α-Gal synthesis and play a role during Anaplasma phagocytophilum infection and Ixodes scapularis tick vector development

Use of Graph Theory to Characterize Human and Arthropod Vector Cell Protein Response to Infection With Anaplasma phagocytophilum

Ixodes scapularis Tick Cells Control Anaplasma phagocytophilum Infection by Increasing the Synthesis of Phosphoenolpyruvate from Tyrosine

Anaplasma phagocytophilum MSP4 and HSP70 Proteins Are Involved in Interactions with Host Cells during Pathogen Infection

Functional Redundancy and Ecological Innovation Shape the Circulation of Tick-Transmitted Pathogens

Protease Inhibitors in Tick Saliva: The Role of Serpins and Cystatins in Tick-host-Pathogen Interaction

Tick-Pathogen Interactions and Vector Competence: Identification of Molecular Drivers for Tick-Borne Diseases

Tick-Pathogen Ensembles: Do Molecular Interactions Lead Ecological Innovation?

Anaplasma phagocytophilum Infection Subverts Carbohydrate Metabolic Pathways in the Tick Vector, Ixodes scapularis

Substrate prediction of Ixodes ricinus salivary lipocalins differentially expressed during Borrelia afzelii infection

Anaplasma phagocytophilum increases the levels of histone modifying enzymes to inhibit cell apoptosis and facilitate pathogen infection in the tick vector Ixodes scapularis

Sialomes and Mialomes: A Systems-Biology View of Tick Tissues and Tick-Host Interactions

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Zobrazit více v PubMed

Dryad
10.5061/dryad.50KT0