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      Immunity against Ixodes scapularis Salivary Proteins Expressed within 24 Hours of Attachment Thwarts Tick Feeding and Impairs Borrelia Transmission

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          Abstract

          In North America, the black-legged tick, Ixodes scapularis, an obligate haematophagus arthropod, is a vector of several human pathogens including Borrelia burgdorferi, the Lyme disease agent. In this report, we show that the tick salivary gland transcriptome and proteome is dynamic and changes during the process of engorgement. We demonstrate, using a guinea pig model of I. scapularis feeding and B. burgdorferi transmission, that immunity directed against salivary proteins expressed in the first 24 h of tick attachment — and not later — is sufficient to evoke all the hallmarks of acquired tick-immunity, to thwart tick feeding and also to impair Borrelia transmission. Defining this subset of proteins will promote a mechanistic understanding of novel I. scapularis proteins critical for the initiation of tick feeding and for Borrelia transmission.

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          Most cited references63

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          Fundamentals of experimental design for cDNA microarrays.

          Microarray technology is now widely available and is being applied to address increasingly complex scientific questions. Consequently, there is a greater demand for statistical assessment of the conclusions drawn from microarray experiments. This review discusses fundamental issues of how to design an experiment to ensure that the resulting data are amenable to statistical analysis. The discussion focuses on two-color spotted cDNA microarrays, but many of the same issues apply to single-color gene-expression assays as well.
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            The biological and social phenomenon of Lyme disease.

            Lyme disease, unknown in the United States two decades ago, is now the most common arthropod-borne disease in the country and has caused considerable morbidity in several suburban and rural areas. The emergence of this disease is in part the consequence of the reforestation of the northeastern United States and the rise in deer populations. Unfortunately, an accurate estimation of its importance to human and animal health has not been made because of difficulties in diagnosis and inadequate surveillance activities. Strategies for prevention of Lyme disease include vector control and vaccines.
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              Amplified RNA synthesized from limited quantities of heterogeneous cDNA.

              The heterogeneity of neural gene expression and the spatially limited expression of many low-abundance messenger RNAs in the brain has made cloning and analysis of such messages difficult. To generate amounts of nucleic acids sufficient for use in standard cloning strategies, we have devised a method for producing amplified heterogeneous populations of RNA from limited quantities of cDNA. Whole cerebellar RNA was primed with a synthetic oligonucleotide containing the T7 RNA polymerase promoter sequence 5' to a polythymidylate region. After second-strand cDNA synthesis, T7 RNA polymerase was used to generate amplified antisense RNA (aRNA). Up to 80-fold molar amplification has been achieved from nanogram quantities of cDNA. The amplified material is similar in size distribution to the parent cDNA and shows sequence heterogeneity as assessed by Southern and Northern blot analysis. Specific messages for moderate-abundance mRNAs for actin and guanine nucleotide-binding protein (G-protein) alpha subunits have been detected in the amplified material. By using in situ transcription to generate cDNA, sequences for cyclophilin have been detected in aRNA derived from single cerebellar tissue sections. cDNA derived from a single cerebellar Purkinje cell also has been amplified and yields material that hybridizes to cognate whole RNA and mRNA but not to Escherichia coli RNA.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS ONE
                plos
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2007
                16 May 2007
                : 2
                : 5
                : e451
                Affiliations
                [1 ]Section of Rheumatology, Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut, United States of America
                [2 ]Section of Allergy and Clinical Immunology, Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut, United States of America
                [3 ]Program in Computational Biology and Bioinformatics, Yale University, New Haven, Connecticut, United States of America
                [4 ]Department of Biochemistry and Biophysics, Yale University, New Haven, Connecticut, United States of America
                [5 ]Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut, United States of America
                [6 ]Department of Entomology, Connecticut Agricultural Experiment Station, New Haven, Connecticut, United States of America
                Massachusetts General Hospital and Harvard Medical School, United States of America
                Author notes
                * To whom correspondence should be addressed. E-mail: erol.fikrig@ 123456yale.edu

                Conceived and designed the experiments: EF FK SN RK BK. Performed the experiments: SN KD NM XL KN JA RK BK. Analyzed the data: EF SN KD TR KN CB JA. Contributed reagents/materials/analysis tools: TR KN. Wrote the paper: EF FK SN JA RK BK.

                Article
                07-PONE-RA-00907R1
                10.1371/journal.pone.0000451
                1866177
                17505544
                0867d527-5e6b-49c5-ab9c-0d6edcb144cf
                Narasimhan et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
                History
                : 14 March 2007
                : 22 April 2007
                Page count
                Pages: 14
                Categories
                Research Article
                Infectious Diseases
                Immunology/Immunity to Infections
                Microbiology/Immunity to Infections
                Microbiology/Medical Microbiology

                Uncategorized
                Uncategorized

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