Transcriptional profiling of the murine cutaneous response during initial and subsequent infestations with Ixodes scapularis nymphs.
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.
Department of Pathology, University of Texas Medical Branch, Galveston, Texas 77555, USA.
Dar M
DM
Heinze
Stephen K
SK
Wikel
Saravanan
S
Thangamani
Francisco J
FJ
Alarcon-Chaidez
D000818:Animals; D002465:Cell Movement; D016207:Cytokines / Q000032:analysis; D015536:Down-Regulation / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000235:genetics*; D054884:Host-Pathogen Interactions / Q000502:physiology*; D006801:Humans; D007113:Immunity, Innate; D007299:Insect Bites and Stings; D018884:Ixodes / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009758:Nymph / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction; D012867:Skin / Q000469:parasitology; D013047:Specific Pathogen-Free Organisms; D013984:Tick Infestations / Q000276:immunology / Q000469:parasitology*; D013997:Time Factors; D015854:Up-Regulation / Q000235:genetics
D016207:Cytokines; D012333:RNA, Messenger