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New Cell Lines Derived from European Tick Species - PubMed

  • ️Sat Jan 01 2022

New Cell Lines Derived from European Tick Species

Lesley Bell-Sakyi et al. Microorganisms. 2022.

Abstract

Tick cell lines are important tools for research on ticks and the pathogens they transmit. Here, we report the establishment of ten new cell lines from European ticks of the genera Argas, Dermacentor, Hyalomma, Ixodes and Rhipicephalus originating from Germany and Spain. For each cell line, the method used to generate the primary culture, a morphological description of the cells and species confirmation by sequencing of the partial 16S rRNA gene are presented. Further molecular analysis of the two new Ixodes ricinus cell lines and three existing cell lines of the same species revealed genetic variation between cell lines derived from ticks collected in the same or nearby locations. Collectively, these new cell lines will support research into a wide range of viral, bacterial and protozoal tick-borne diseases prevalent in Europe.

Keywords: Argas reflexus; Dermacentor reticulatus; Hyalomma lusitanicum; Hyalomma scupense; Ixodes ricinus; Rhipicephalus bursa; tick cell line; vector.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1

Argas reflexus and Dermacentor reticulatus cell lines. (a) A. reflexus primary culture 4 months post initiation showing large floating vacuolated cells, smaller attached spindle-shaped cells and small round haemocyte-like cells; live, inverted microscope; (b) A. reflexus cell line ARE/LULS41 at passage 8, 5 years post initiation; live, inverted microscope; (c) cluster of floating vacuolated cells in D. reticulatus primary culture, 15 months post initiation, showing pink colouring of some vacuoles; (d) D. reticulatus cell line DRE/LULS60 at passage 7, 3 years post initiation. Live, inverted microscope, scale bars = 100 µm.

Figure 2
Figure 2

Hyalomma lusitanicum and Hyalomma scupense cell lines. (a) H. lusitanicum cell line HLE/LULS42 at passage 2, 8 years after initiation; (b) H. lusitanicum cell line HLE/LULS48 at passage 6, 8 years after initiation, showing a cell cluster bounded by a membrane (arrow); (c) H. scupense cell line HSE/LULS51 at passage 9, 6 years after initiation; (d) H. scupense cell line HSE/LULS59 at passage 9, 6 years after initiation, showing extracellular matrix associated with the multicellular vesicles (arrows). Live, inverted microscope, scale bars = 100 µm.

Figure 3
Figure 3

Ixodes ricinus and Rhipicephalus bursa cell lines. (a) I. ricinus cell line IRE/LUAP46 at passage 3, 7 years after initiation; (b) I. ricinus cell line IRE/LULS55 at passage 19, 44 months after initiation; (c) R. bursa cell line RBE/LULS58 at passage 3, 4 years after initiation. Live, inverted microscope, scale bars = 100 µm.

Figure 4
Figure 4

Maximum likelihood phylogenies of 16s rRNA and trospA gene fragments amplified from the Ixodes ricinus cell lines IRE/LUAP46, IRE/LULS55, IRE/CTVM19, IRE/CTVM20 and IRE11. (a) 16s rRNA (441 nucleotide positions): sequences in the 184-AG haplogroup shown in red and sequences in 184-CT haplogroup shown in blue. (b) trospA (695 nucleotide positions): sequences from North Africa and Portugal shown in turquoise and sequences from Europe, excluding central and southern Portugal, shown in purple. Sequences generated from the I. ricinus cell lines are in bold. Sequences obtained from NCBI GenBank are represented by the accession numbers and followed by the country codes. Ir = I. ricinus, Ii = Ixodes inopinatus, I = Ixodes sp. Bootstrap support values greater than 70% are shown adjacent to the nodes. Scale bars represent the number of substitutions per site.

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