Transcriptional profiling of the murine cutaneous response during initial and subsequent infestations with Ixodes scapularis nymphs

Particle and Fibre Toxicology

Francisco J Alarcon-Chaidez²  Saravanan Thangamani¹  Stephen K Wikel³  Dar M Heinze² 

[1] Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, Texas 77555, USA;Department of Pathology, University of Texas Medical Branch, Galveston, Texas 77555, USA;Department of Basic Medical Sciences, Quinnipiac University School of Medicine, Hamden, CT 06518, USA

关键词: Gene expression profiling;    Tick-host interface;    Cutaneous response;    Skin;    Ixodes scapularis;    Tick(s);   

Others  :  1233099
DOI  :  10.1186/1756-3305-5-26

 received in 2011-11-02, accepted in 2012-02-06,  发布年份 2012

【 摘 要 】

Background

Ixodes scapularis ticks are hematophagous arthropods capable of transmitting many infectious agents to humans. The process of blood feeding is an extended and continuous interplay between tick and host responses. While this process has been studied extensively in vitro, no global understanding of the host response to ticks has emerged.

Methods

To address this issue, we used PCR-arrays to measure skin-specific expression of 233 discrete genes at 8 time points during primary and secondary infestations of mice with pathogen-free I. scapularis nymphs. Selected results were then validated at the mRNA and protein levels by additional real-time PCR and bioplex assay.

Results

Primary infestation was characterized by the late induction of an innate immune response. Lectin pattern recognition receptors, cytokines, and chemokines were upregulated consistent with increased neutrophil and macrophage migration. Gene ontology and pathway analyses of downregulated genes suggested inhibition of gene transcription and Th17 immunity. During the secondary infestation, additional genes were modulated suggesting a broader involvement of immune cells including CD8 and CD4 positive T lymphocytes. The cytokine response showed a mixed Th1/Th2 profile with a potential for T regulatory cell activity. Key gene ontology clusters observed during the secondary infestation were cell migration and activation. Matrix metalloproteinases were upregulated, apoptosis-related genes were differentially modulated, and immunoreceptor signaling molecules were upregulated. In contrast, transcripts related to mitogenic, WNT, Hedgehog, and stress pathways were downregulated.

Conclusions

Our results support a model of tick feeding where lectin pattern recognition receptors orchestrate an innate inflammatory response during primary infestation that primes a mixed Th1/Th2 response upon secondary exposure. Tick feeding inhibits gene transcription and Th17 immunity. Salivary molecules may also inhibit upregulation of mitogenic, WNT, Hedgehog, and stress pathways and enhance the activity of T regulatory cells, production of IL-10, and suppressors of cytokine signaling molecules (SOCS). This study provides the first comprehensive transcriptional analysis of the murine host response at the I. scapularis bite site and suggests both a potential model of the host cutaneous response and candidate genes for further description and investigation.

【 授权许可】

   
2012 Heinze et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Brossard M, Wikel S: Tick Immunobiology. In Ticks: Biology, Disease and Control. Edited by Bowman AS, Nuttall PA. Cambridge, UK: Cambridge University Press; 2008:186-204.
• [2]Wikel SK, Ramachandra RN, Bergman DK, Burkot TR, Piesman J: Infestation with pathogen-free nymphs of the tick Ixodes scapularis induces host resistance to transmission of Borrelia burgdorferi by ticks. Infect Immun 1997, 65:335-338.
• [3]Trager W: Acquired immunity to ticks. Journal of Parasitology 1939, 25:57-81.
• [4]Muller-Doblies UU, Maxwell SS, Boppana VD, Mihalyo MA, McSorley SJ, Vella AT, Adler AJ, Wikel SK: Feeding by the tick, Ixodes scapularis, causes CD4(+) T cells responding to cognate antigen to develop the capacity to express IL-4. Parasite Immunol 2007, 29:485-499.
• [5]Anguita J, Ramamoorthi N, Hovius JW, Das S, Thomas V, Persinski R, Conze D, Askenase PW, Rincon M, Kantor FS, Fikrig E: Salp15, an ixodes scapularis salivary protein, inhibits CD4(+) T cell activation. Immunity 2002, 16:849-859.
• [6]Hovius JW, de Jong MA, den Dunnen J, Litjens M, Fikrig E, van der Poll T, Gringhuis SI, Geijtenbeek TB: Salp15 binding to DC-SIGN inhibits cytokine expression by impairing both nucleosome remodeling and mRNA stabilization. PLoS Pathog 2008, 4:e31.
• [7]Guo X, Booth CJ, Paley MA, Wang X, DePonte K, Fikrig E, Narasimhan S, Montgomery RR: Inhibition of neutrophil function by two tick salivary proteins. Infect Immun 2009, 77:2320-2329.
• [8]Oliveira CJ, Cavassani KA, More DD, Garlet GP, Aliberti JC, Silva JS, Ferreira BR: Tick saliva inhibits the chemotactic function of MIP-1alpha and selectively impairs chemotaxis of immature dendritic cells by down-regulating cell-surface CCR5. Int J Parasitol 2008, 38:705-716.
• [9]Wikel SK: Host immunity to ticks. Annu Rev Entomol 1996, 41:1-22.
• [10]Kongsuwan K, Piper EK, Bagnall NH, Ryan K, Moolhuijzen P, Bellgard M, Lew A, Jackson L, Jonsson NN: Identification of genes involved with tick infestation in Bos taurus and Bos indicus. Dev Biol (Basel) 2008, 132:77-88.
• [11]Piper EK, Jackson LA, Bagnall NH, Kongsuwan KK, Lew AE, Jonsson NN: Gene expression in the skin of Bos taurus and Bos indicus cattle infested with the cattle tick, Rhipicephalus (Boophilus) microplus. Vet Immunol Immunopathol 2008, 126:110-119.
• [12]Piper EK, Jonsson NN, Gondro C, Lew-Tabor AE, Moolhuijzen P, Vance ME, Jackson LA: Immunological profiles of Bos taurus and Bos indicus cattle infested with the cattle tick, Rhipicephalus (Boophilus) microplus. Clin Vaccine Immunol 2009, 16:1074-1086.
• [13]Nestle FO, Di Meglio P, Qin JZ, Nickoloff BJ: Skin immune sentinels in health and disease. Nat Rev Immunol 2009, 9:679-691.
• [14]Bouchard KR, Wikel SK: Care, maintenance, and experimental infestation of ticks in the laboratory setting. In Biology of Disease Vectors. Second edition. Edited by Marquart WC. San Diego: Elsevier Academic Press; 2005:705-711.
• [15]Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002, 3:RESEARCH0034.
• [16]Dvinge H, Bertone P: HTqPCR: high-throughput analysis and visualization of quantitative real-time PCR data in R. Bioinformatics 2009, 25:3325-3326.
• [17]Huang da W, Sherman BT, Lempicki RA: Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists. Nucleic Acids Res 2009, 37:1-13.
• [18]Huang da W, Sherman BT, Lempicki RA: Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 2009, 4:44-57.
• [19]Maitra A, Shen F, Hanel W, Mossman K, Tocker J, Swart D, Gaffen SL: Distinct functional motifs within the IL-17 receptor regulate signal transduction and target gene expression. Proc Natl Acad Sci USA 2007, 104:7506-7511.
• [20]Torcia MG, Santarlasci V, Cosmi L, Clemente A, Maggi L, Mangano VD, Verra F, Bancone G, Nebie I, Sirima BS, et al.: Functional deficit of T regulatory cells in Fulani, an ethnic group with low susceptibility to Plasmodium falciparum malaria. Proc Natl Acad Sci USA 2008, 105:646-651.
• [21]Brown GD: Dectin-1: a signalling non-TLR pattern-recognition receptor. Nat Rev Immunol 2006, 6:33-43.
• [22]Kopecky J, Kuthejlova M, Pechova J: Salivary gland extract from Ixodes ricinus ticks inhibits production of interferon-gamma by the upregulation of interleukin-10. Parasite Immunol 1999, 21:351-356.
• [23]Schoeler GB, Manweiler SA, Wikel SK: Ixodes scapularis: effects of repeated infestations with pathogen-free nymphs on macrophage and T lymphocyte cytokine responses of BALB/c and C3H/HeN mice. Exp Parasitol 1999, 92:239-248.
• [24]Kovar L, Kopecky J, Rihova B: Salivary gland extract from Ixodes ricinus tick polarizes the cytokine profile toward Th2 and suppresses proliferation of T lymphocytes in human PBMC culture. J Parasitol 2001, 87:1342-1348.
• [25]Schoeler GB, Manweiler SA, Wikel SK: Cytokine responses of C3H/HeN mice infested with Ixodes scapularis or Ixodes pacificus nymphs. Parasite Immunol 2000, 22:31-40.
• [26]Dinarello CA: Immunological and inflammatory functions of the interleukin-1 family. Annu Rev Immunol 2009, 27:519-550.
• [27]Leeman MF, Curran S, Murray GI: The structure, regulation, and function of human matrix metalloproteinase-13. Crit Rev Biochem Mol Biol 2002, 37:149-166.
• [28]Strieter RM, Burdick MD, Gomperts BN, Belperio JA, Keane MP: CXC chemokines in angiogenesis. Cytokine Growth Factor Rev 2005, 16:593-609.
• [29]Viola A, Luster AD: Chemokines and their receptors: drug targets in immunity and inflammation. Annu Rev Pharmacol Toxicol 2008, 48:171-197.
• [30]Krause PJ, Grant-Kels JM, Tahan SR, Dardick KR, Alarcon-Chaidez F, Bouchard K, Visini C, Deriso C, Foppa IM, Wikel S: Dermatologic changes induced by repeated Ixodes scapularis bites and implications for prevention of tick-borne infection. Vector Borne Zoonotic Dis 2009, 9:603-610.
• [31]Mulenga A, Khumthong R, Chalaire KC: Ixodes scapularis tick serine proteinase inhibitor (serpin) gene family; annotation and transcriptional analysis. BMC Genomics 2009, 10:217. BioMed Central Full Text
• [32]Prevot PP, Couvreur B, Denis V, Brossard M, Vanhamme L, Godfroid E: Protective immunity against Ixodes ricinus induced by a salivary serpin. Vaccine 2007, 25:3284-3292.
• [33]Sternlicht MD, Werb Z: How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol 2001, 17:463-516.
• [34]Francischetti IM, Mather TN, Ribeiro JM: Tick saliva is a potent inhibitor of endothelial cell proliferation and angiogenesis. Thromb Haemost 2005, 94:167-174.
• [35]Maxwell SS, Stoklasek TA, Dash Y, Macaluso KR, Wikel SK: Tick modulation of the in-vitro expression of adhesion molecules by skin-derived endothelial cells. Ann Trop Med Parasitol 2005, 99:661-672.
• [36]Brown SJ, Galli SJ, Gleich GJ, Askenase PW: Ablation of immunity to Amblyomma americanum by anti-basophil serum: cooperation between basophils and eosinophils in expression of immunity to ectoparasites (ticks) in guinea pigs. J Immunol 1982, 129:790-796.
• [37]Wada T, Ishiwata K, Koseki H, Ishikura T, Ugajin T, Ohnuma N, Obata K, Ishikawa R, Mukai K, Yoshikawa S, Kawano Y, Minegishi Y, Yokozeki H, Watanabe N, Karasuyama H: Selective ablation of basophils in mice reveals their nonredundant role in acquired immunity against ticks. J Clin Invest 2010, 120:2867-2875.
• [38]Kim S, Prout M, Ramshaw H, Lopez AF, LeGros G, Min B: Cutting edge: basophils are transiently recruited into the draining lymph nodes during helminth infection via IL-3, but infection-induced Th2 immunity can develop without basophil lymph node recruitment or IL-3. J Immunol 2010, 184:1143-1147.
• [39]Barlow JL, McKenzie AN: IL-25: a key requirement for the regulation of type-2 immunity. Biofactors 2009, 35:178-182.
• [40]Xu M, Mizoguchi I, Morishima N, Chiba Y, Mizuguchi J, Yoshimoto T: Regulation of antitumor immune responses by the IL-12 family cytokines, IL-12, IL-23, and IL-27. Clin Dev Immunol 2010., 2010
• [41]McLaren JE, Ramji DP: Interferon gamma: a master regulator of atherosclerosis. Cytokine Growth Factor Rev 2009, 20:125-135.
• [42]Dash Y, Maxwell SS, Rajan TV, Wikel SK: Murine extramedullary erythropoiesis induced by tick infestation. Ann Trop Med Parasitol 2005, 99:518-531.
• [43]Starnes T, Broxmeyer HE, Robertson MJ, Hromas R: Cutting edge: IL-17D, a novel member of the IL-17 family, stimulates cytokine production and inhibits hemopoiesis. J Immunol 2002, 169:642-646.
• [44]Ramachandra RN, Wikel SK: Modulation of host-immune responses by ticks (Acari: Ixodidae): effect of salivary gland extracts on host macrophages and lymphocyte cytokine production. J Med Entomol 1992, 29:818-826.
• [45]Sa-Nunes A, Bafica A, Antonelli LR, Choi EY, Francischetti IM, Andersen JF, Shi GP, Chavakis T, Ribeiro JM, Kotsyfakis M: The immunomodulatory action of sialostatin L on dendritic cells reveals its potential to interfere with autoimmunity. J Immunol 2009, 182:7422-7429.
• [46]Li WX: Canonical and non-canonical JAK-STAT signaling. Trends Cell Biol 2008, 18:545-551.
• [47]Hayden MS, Ghosh S: Shared principles in NF-kappaB signaling. Cell 2008, 132:344-362.
• [48]Bikkavilli RK, Malbon CC: Mitogen-activated protein kinases and Wnt/beta-catenin signaling: Molecular conversations among signaling pathways. Commun Integr Biol 2009, 2:46-49.
• [49]Ingham PW, Nakano Y, Seger C: Mechanisms and functions of Hedgehog signalling across the metazoa. Nat Rev Genet 2011, 12:393-406.