Molecular Characterization and Related Aspects of the Innate Immune Response in Ticks

Compared to insects, little is known about innate immunity in ticks. This chapter addresses the molecular processes that recognize non-self and the cellular and molecular processes mobilized to phagocytose, engulf, inhibit or kill invaders. We discuss the receptors that recognize pathogen associated molecular patterns (PAMPs) and the putative up-regulation of regulatory cascades that lead, ultimately, to cellular or molecular responses. We describe the molecular events that activate the cellular processes and the array of humoral factors that are mobilized against invading organisms, including antimicrobial peptides, proteases and protease inhibitors, lectins, coagulation factors and others. Special attention is directed to the antimicrobial activity of the midgut, the initial site of contact for microbes ingested with the blood. Blood feeding and digestion alone up-regulates an impressive array of proteins, e.g. oxidative stress reducing proteins, lectins, protease inhibitors, proteases, hydrolases, protein/lipid binding agents. Finally, we compare the innate immune responses of ticks with insects and other invertebrates and note deficiencies in our knowledge tick innate immunity

Hyaluronidases of Gram-Positive Bacteria

Bacterial hyaluronidases, enzymes capable of breaking down hyaluronate, are produced by a number of pathogenic Gram-positive bacteria that initiate infections at the skin or mucosal surfaces. Since reports of the hyaluronidases first appeared, there have been numerous suggestions as to the role of the enzyme in the disease process. Unlike some of the other more well studied virulence factors, much of the information on the role of hyaluronidase is speculative, with little or no data to substantiate proposed roles. Over the last 5 years, a number of these enzymes from Gram-positive organisms have been cloned, and the nucleotide sequence determined. Phylogenetic analysis, using the deduced amino acid sequences of the Gram-positive hyaluronidases, suggests a relatedness among some of the enzymes. Molecular advances may lend to a more thorough understanding of the role of hyaluronidases in bacterial physiology and pathogenesis. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved

Prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii in Amblyomma americanum and Dermacentor variabilis Collected From Southeastern Virginia, 2010-2011

Amblyomma americanum is the most commonly-encountered tick species in southeastern Virginia, representing approximately 95% of the human-biting tick population in this area. Here we investigated the prevalence of Ehrlichia chaffeensis and Ehrlichia ewingii in questing Amblyomma americanum and Dermacentor variabilis ticks collected from multiple sites in southeastern Virginia from 2010–2011. Although both Ehrlichia species were detected in Amblyomma americanum, no evidence of either pathogen was found in Dermacentor variabilis. Prevalence of E. chaffeensis varied by location, ranging from 0 – 5.08% among Amblyomma americanum populations. Ehrlichia ewingii prevalence was slightly higher, ranging from 0 – 8.20% among A. americanum populations. We conclude that both pathogens are established in southeastern Virginia A. americanum populations, and that although there are no apparent temporal trends in Ehrlichia prevalence, there is variation among locations, suggesting the potential for disease hotspots

Identification of a Defensin from the Hemolymph of the American Dog Tick, Dermacentor variabilis

Hemolymph from partially fed virgin Dermacentor variabilis females was collected following Borrelia burgdorferi challenge and assayed for antimicrobial activity against Bacillus subtilis and B. burgdorferi. A small inducible cationic peptide was identified by SDS-PAGE in the hemolymph of these ticks as early as 1h post challenge. Following purification by a three-step procedure involving sequential SepPak elution, reversed phase high performance liquid chromatography (RP-HPLC) and gel electrophoresis, the yield of the active peptide was approximately 0.1% of the total protein in the hemolymph plasma. The molecular weight, 4.2kDa, was determined by MALDI-TOF mass spectrometry. N-terminal sequencing by the Edman degradation method gave a sequence for the first 30 amino acids as: G-F-G-C-P-L-N-Q-G-A-C-H-N-H-C-R-S-I-(R)-(R)-(R)-G-G-Y-C-S-Q-I-I-K. A computer search of databases showed that the peptide had 83% similarity to a defensin found in a scorpion. This is the first report of a defensin from a tick. The peptide was stable at least up to 70 degree C. Although the tick defensin alone was not immediately effective against B. burgdorferi, tick defensin plus lysozyme killed more than 65% of cultured B. burgdorferi within 1h

Response of the Tick Dermacentor variabilis (Acari: Ixodidae) to Hemocoelic Inoculation of Borrelia burgdorferi (Spirochetales)

When Borrelia burgdorferi B31 low passage strain spirochetes were directly injected into the hemocoel of Dermacentor variabilis(Say) females, the bacteria were cleared from the hemocoel within \u3c 24 h. Viable spirochetes were not found in hemolymph, salivary gland, or ovary tissues by subculturing or by IFA. The hemocyte population increased ≈6 times within the first 6 h after inoculation compared with the uninoculated controls. In contrast, the soluble total hemolymph protein content decreased inversely with the increase in hemocytes. Borreliacidal activity was demonstrated with cell-free hemolymph from D. variabilis. In vitro antimicrobial assays using hemolymph from borrelia-challenged and nonchallenged ticks resulted in 72% spirochete reductions compared with only 11.5%, respectively, within 1 h. Additional evidence of induced antimicrobial hemolymph protein activity was demonstrated by SDS-PAGE, which revealed upregulation of a lysozyme-like peptide (≈ 15 kDa) (22% increase) and the induction of a ≈ 5.8 kDa peptide in the B. burgdorferi-challenged ticks. In contrast with the nonvector borne Bacillus subtilis, D. variabilis presented a rapid and robust response to challenge with cultured B. burgdorferi spirochetes and lead to their early elimination. The role of the tick immune system, including possible differences between vector and nonvector ticks, in determining the success of invasive bacteria is discussed

Resistance of the Tick Dermacentor variabilis (Acari: Ixodidae) Following Challenge with the Bacterium Escherichia coli (Enterobacteriales: Enterobacteriaceae)

In addition to a soluble response, many invertebrates control bacterial infections by means of phagocytosis or melanotic encapsulation. In some insects, Escherichia coli growth is reported to be inhibited by aggregation/encapsulation. Soluble and phagocytic responses to bacterial challenge have been reported in ticks, but evidence of an aggregation/encapsulation response was reported only for inanimate (araldite) implants. Ticks were challenged by direct inoculation of bacteria into the hemocoel cavity. By plating, no viable E. coli were detected 6 h postinoculation. A direct fluorescence assay (DFA) revealed aggregated bacteria 1 h postinoculation. Furthermore, DFA showed aggregated bacteria at 6, 24, and 48 h postinoculation associated with masses of tissue, presumably of cellular origin, suggesting events similar to those described as nodulation. These findings suggest that encapsulation/nodulation may be an important component of the immune response in ticks

Tissue and Life Stage Distribution of a Defensin-Like Gene in the Lone Star Tick Amblyomma americanum

The transcript sequence of the Amblyomma americanum Linnaeus (Acari: Ixodidae) defensin, termed amercin (amn), was ascertained and a 219-bp amn coding region identified. The gene encodes a 72-amino acid prepropeptide with a putative 37-amino acid mature peptide. This gene shows little similarity to either of the defensins from Amblyomma hebraeum Koch, the only other Amblyomma species for which a defensin has been described. Sequence comparisons with other tick defensins reveal amn to be shorter (6 bp or 2 amino acids) than the Ixodes scapularis Linnaeus (Acari: Ixodidae) and Dermacentor variabilis (Say) (Acari: Ixodidae) defensin sequences. The amercin prepropeptide has 60.8% and 59.5% similarity with the I. scapularis and D. variabilis prepropeptides, respectively, whereas the mature amercin peptide has 73.7% and 71.1% similarity with the mature peptides of these ticks. Similarity with other tick defensins ranges from 42% to 71%. In A. americanum, defensin transcript was found in the midgut, fat body and salivary gland tissues, as well as in the haemocytes. Defensin transcript was also present in early-stage eggs (less than 48 h old), late-stage eggs (approximately 2 weeks old), larvae and nymphs of A. americanum and I. scapularis, both of which are vector-competent for Borrelia spirochetes

Nucleotide Sequence of the Streptococcin A-FF22 Lantibiotic Regulon: Model for Production of the Lantibiotic SA-FF22 by Strains of Streptococcus pyogenes

Streptococcin A-FF22 (SA-FF22) is a type AII linear lantibiotic produced by Streptococcus pyogenes strain FF22. Sequence analysis of an approximate 10 kb region of DNA showed it to contain nine open reading frames arranged in three operons responsible for regulation, biosynthesis and immunity of SA-FF22. This region is organized similarly to the Lactococcus lactis lacticin 481 region, however, unlike lacticin 481, a two-component regulatory system is essential for SA-FF22 production. located immediately downstream of the scn region is a putative transposase gene, the presence of which supports earlier data that indicated a mobile nature to this region. (©) 1999 Published by Elsevier Science B.V. All rights reserved