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Comparative Study
, 12 (1), 212

Tissue-specific Signatures in Tick Cell Line MS Profiles

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Comparative Study

Tissue-specific Signatures in Tick Cell Line MS Profiles

Dmitry S Loginov et al. Parasit Vectors.

Abstract

Background: The availability of tick in vitro cell culture systems has facilitated many aspects of tick research, including proteomics. However, certain cell lines have shown a tissue-specific response to infection. Thus, a more thorough characterization of tick cell lines is necessary. Proteomic comparative studies of various tick cell lines will contribute to more efficient application of tick cell lines as model systems for investigation of host-vector-pathogen interactions.

Results: Three cell lines obtained from a hard tick, Ixodes ricinus, and two from I. scapularis were investigated. A cell mass spectrometry approach (MALDI-TOF MS) was applied, as well as classical proteomic workflows. Using PCA, tick cell line MS profiles were grouped into three clusters comprising IRE/CTVM19 and ISE18, IRE11 and IRE/CTVM20, and ISE6 cell lines. Two other approaches confirmed the results of PCA: in-solution digestion followed by nanoLC-ESI-Q-TOF MS/MS and 2D electrophoresis. The comparison of MS spectra of the cell lines and I. ricinus tick organs revealed 29 shared peaks. Of these, five were specific for ovaries, three each for gut and salivary glands, and one for Malpighian tubules. For the first time, characteristic peaks in MS profiles of tick cell lines were assigned to proteins identified in acidic extracts of corresponding cell lines.

Conclusions: Several organ-specific MS signals were revealed in the profiles of tick cell lines.

Keywords: Biotyping; MALDI-TOF MS; Tick; Tick cell line; Tick organs.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Positive ion MALDI-TOF mass spectra of the IRE/CTVM19 tick cell line. Matrices applied by the dried droplet method were: a CHCA (20 mg/ml), b DHB (20 mg/ml), c FA (10 mg/ml), and d SA (10 mg/ml) in acetonitrile/2.5% v/v trifluoroacetic acid, 7:3 (v/v)
Fig. 2
Fig. 2
Combining matrix compounds to enhance the signal intensity and the number of peaks in peptide/protein profiles. MS profiles of the IRE/CTVM19 tick cell line were obtained after mixing FA and SA in ratios of: a 10:5, b 5:5, c 5:10 and d 5:15 mg/ml
Fig. 3
Fig. 3
Principal component analysis from the MS spectra of tick cell lines. a PCA three-dimensional plot from MS spectra of tick cell lines: IRE11 (red dots); IRE/CTVM19 (green dots); IRE/CTVM20 (blue dots); ISE6 (yellow dots); and ISE18 (purple dots). MS profiles of tick cell lines formed three clusters: IRE/CTVM19 and ISE18, IRE11 and IRE/CTVM20, and ISE6. b 3D factor loadings plot. Factors with loadings > 0.2 are marked in blue
Fig. 4
Fig. 4
Principal component analysis from the MS spectra of tick cell lines and tick organs. a PCA three-dimensional image from MS spectra of tick cell lines and tick organs: IRE11 (red dots); IRE/CTVM19 (light green dots); IRE/CTVM20 (light blue dots); ISE6 (yellow dots); ISE18 (purple dots); gut (deep green); salivary glands (brown); ovaries (magenta); and Malpighian tubules (deep blue). Distinct clusters were formed by MS profiles of tick cell lines and organs. b 3D factor loadings plot. Factors with loadings > 0.2 are marked in blue
Fig. 5
Fig. 5
Tricine-SDS-PAGE gel with separated peptide/protein ACN/TFA extracts from tick cell lines. Lane M: marker; Lane 1: IRE11; Lane 2: IRE/CTVM19; Lane 3: IRE/CTVM20; Lane 4: ISE6; Lane 5: ISE18. Numbered gel segments (labeled as fr) were processed for in-gel digestion and further analyses
Fig. 6
Fig. 6
Two-dimensional electrophoresis of tick cell line protein extracts. a IRE11. b IRE/CTVM19. c IRE/CTVM20. d ISE6. e ISE18. Gels were compared within IRE and ISE groups using the SameSpots software, and protein spots with significantly different expression (ANOVA, P < 0.05) were excised from the gels for MALDI-TOF/TOF MS/MS (marked in blue). Protein ladder: 245, 180, 135, 100, 75, 63, 48, 35, 25, 20, 17, 11 kDa

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