Expression Profiles of Toll-Like Receptors in the Differentiation of an Infection with Borrelia burgdorferi Sensu Lato Spirochetes.
The similarity of Lyme borreliosis to other diseases and its complex pathogenesis present diagnostic and therapeutic difficulties. The changes that occur at the cellular and molecular levels after a Borrelia sp. infection still remain poorly understood. Therefore, the present study focused on the expression of TLR and TLR-signaling genes in human dermal fibroblasts in the differentiation of an infection with Borrelia burgdorferi sensu lato spirochetes. Normal human dermal fibroblasts were cultured with the spirochetes of Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii. Total RNA was extracted from the cells using TRIzol reagent. The analysis of the expression profiles of TLRs and TLR-related genes was performed using commercially available oligonucleotide microarrays of HG-U133A. The GeneSpring 12.0 platform and significance analysis of microarrays were used for the statistical analysis of microarray data. The analyses using the oligonucleotide microarray and QRT-PCR techniques permitted to identify the genes encoding TLR4 and TLR6 as specific for infection with B. afzelii and B. burgdorferi sensu stricto. In turn, TLR3 was only characteristic for an infection with B. burgdorferi sensu stricto. There were no changes in the TLR gene expression after infection with B. garinii. Our findings confirm that Borrelia has a major effect on fibroblast gene expression. Further characterization of changes in gene expression may lead to valuable insights into the role of the toll-like receptor in the pathogenesis of Lyme disease and may provide guidelines for the development of diagnostic markers for an infection with a particular Borrelia genospecies. Moreover, this will help to identify better treatment strategies for Lyme disease.
Department of Pharmacognosy and Phytochemistry, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Sosnowiec, Poland.
Slawomir
S
Dudek
Department of Internal Medicine, School of Public Health, Silesian Medical University in Katowice, Bytom, Poland.
Ewa
E
Ziółko
Department of Food and Nutrition, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Sosnowiec, Poland.
Magdalena
M
Kimsa-Dudek
Department of Parasitology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Sosnowiec, Poland.
Krzysztof
K
Solarz
Department of Molecular Biology, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Sosnowiec, Poland.
Urszula
U
Mazurek
Faculty of Energy and Environmental Engineering, Silesian University of Technology, Gliwice, Poland.
Aleksander
A
Wierzgoń
Department of Internal Medicine, School of Public Health, Silesian Medical University in Katowice, Bytom, Poland.
Teresa
T
Kokot
Department of Internal Medicine, School of Public Health, Silesian Medical University in Katowice, Bytom, Poland. mwierzgon@sum.edu.pl.
Małgorzata
M
Muc-Wierzgoń
D025065:Borrelia burgdorferi; D015748:Borrelia burgdorferi Group; D002454:Cell Differentiation; D005347:Fibroblasts / Q000378:metabolism / Q000382:microbiology*; D006801:Humans; D008193:Lyme Disease / Q000382:microbiology*; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000737:chemistry; D015398:Signal Transduction; D012867:Skin / Q000378:metabolism; D051193:Toll-Like Receptors / Q000378:metabolism*; D059467:Transcriptome
D009841:Oligonucleotides; D051193:Toll-Like Receptors
Borrelia burgdorferi sensu lato spirochetes; Fibroblasts cultures; Oligonucleotide microarray; QRT-PCR; Toll-like receptors