Abstract

BackgroundFibrinogen-related proteins with lectin activity are believed to be part of the tick innate immune system. Several fibrinogen-related proteins have been described and characterised mainly on the basis of their cDNA sequences while direct biochemical evidence is missing. One of them, the haemolymph lectin Dorin M from the tick Ornithodoros moubata was isolated and characterised in more depth.ResultsSeveral fibrinogen-related proteins were detected in the haemolymph of ixodid ticks Dermacentor marginatus, Rhipicephalus appendiculatus, R. pulchellus, and R. sanguineus. These proteins were recognised by sera directed against the tick lectin Dorin M and the haemagglutination activity of the ticks R. appendiculatus and D. marginatus. Cross-reactivity of the identified proteins with antibodies against the fibrinogen domain of the human ficolin was also shown. The carbohydrate-binding ability of tick haemolymph was confirmed by haemagglutination activity assays, and this activity was shown to be inhibited by neuraminic acid and sialylated glycoproteins as well as by N-acetylated hexosamines. The fibrinogen-related proteins were shown to be glycosylated and they were localised in salivary glands, midguts, and haemocytes of D. marginatus. Hemelipoglycoprotein was also recognised by sera directed against the fibrinogen-related proteins in all three Rhipicephalus species as well as in D. marginatus. However, this protein does not contain the fibrinogen domain and thus, the binding possibly results from the structure similarity between hemelipoglycoprotein and the fibrinogen domain.ConclusionsThe presence of fibrinogen-related proteins was shown in the haemolymph of four tick species in high abundance. Reactivity of antibodies directed against ficolin or fibrinogen-related proteins with proteins which do not contain the fibrinogen domain points out the importance of sequence analysis of the identified proteins in further studies. Previously observed expression of fibrinogen-related proteins in haemocytes together with the results of this study suggest involvement of fibrinogen-related proteins in tick immunity processes. Thus, they have potential as targets for anti-tick vaccines and as antimicrobial proteins in pharmacology. Research on fibrinogen-related proteins could reveal further details of tick innate immunity processes.

Highlights

  • Fibrinogen-related proteins with lectin activity are believed to be part of the tick innate immune system

  • Tick haemolymph haemagglutination Haemagglutination activity (HA) analyses of tick haemolymph samples isolated from Dermacentor marginatus, Rhipicephalus appendiculatus, R. pulchellus, and R. sanguineus were performed using 2% suspension of rabbit erythrocytes

  • Differences in HA inhibition were detected among tick species and the highest inhibition was observed in R. sanguineus

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Summary

Introduction

Fibrinogen-related proteins with lectin activity are believed to be part of the tick innate immune system. Several fibrinogen-related proteins have been described and characterised mainly on the basis of their cDNA sequences while direct biochemical evidence is missing. Tick haemolymph is a complex fluid composed of plasma and haemocytes. It serves several functions such as transportation of nutrients but it contains the. Invertebrate/arthropod lectins are believed to be functional analogues of immunoglobulins due to their specific binding to surface carbohydrate structures of pathogens [11]. Fibrinogen-related proteins (FRePs) are described as humoral factors of the innate immune system with the ability to recognise PAMPs. FRePs are molecules containing fibrinogen-related domain in the

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