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1.
Mol Microbiol ; 115(6): 1395-1409, 2021 06.
Article in English | MEDLINE | ID: mdl-33512032

ABSTRACT

Lyme borreliosis is a tick-borne disease caused by Borrelia burgdorferi sensu lato spirochetes (Lyme borreliae). When the disease affects the central nervous system, it is referred to as neuroborreliosis. In Europe, neuroborreliosis is most often caused by Borrelia garinii. Although it is known that in the host Lyme borreliae spread from the tick bite site to distant tissues via the blood vasculature, the adherence of Lyme borreliae to human brain microvascular endothelial cells has not been studied before. Decorin binding proteins are adhesins expressed on Lyme borreliae. They mediate the adhesion of Lyme borreliae to decorin and biglycan, and the lysine residues located in the binding site of decorin binding proteins are important to the binding activity. In this study, we show that lysine residues located in the canonical binding site can also be found in decorin binding proteins of Borrelia garinii, and that these lysines contribute to biglycan and decorin binding. Most importantly, we show that the lysine residues are crucial for the binding of Lyme borreliae to decorin and biglycan expressing human brain microvascular endothelial cells, which in turn suggests that they are involved in the pathogenesis of neuroborreliosis.


Subject(s)
Adhesins, Bacterial/metabolism , Bacterial Adhesion/physiology , Biglycan/metabolism , Borrelia burgdorferi Group/metabolism , Decorin/metabolism , Lyme Neuroborreliosis/pathology , Adhesins, Bacterial/genetics , Amino Acid Sequence , Binding Sites/genetics , Borrelia burgdorferi Group/genetics , Brain/blood supply , Cells, Cultured , Endothelial Cells/metabolism , Humans , Lyme Neuroborreliosis/microbiology , Lysine/chemistry , Molecular Dynamics Simulation , Sequence Alignment , Tick-Borne Diseases/microbiology
2.
J Microbiol Methods ; 180: 106122, 2021 01.
Article in English | MEDLINE | ID: mdl-33326821

ABSTRACT

The cut-off values used in C6 peptide-based enzyme immunoassay (EIA), a widely used test in Lyme borreliosis (LB) serology, have not been thoroughly analysed. The objective of the study was to examine the performance of the C6 EIA, and to determine optimal cut-off values for the test. The analysed data contained results of 1368 serum samples. C6 EIA index values were compared statistically with the immunoblot (IB) test results. The identified cut-off values were further tested in a well-defined LB patient cohort. Cut-off value 1.6 appeared to be optimal when C6 EIA was used as a stand-alone test. When using C6 EIA as the first-tier test, the optimal cut-off values were 0.9 and 2.4 for negative and positive results. When C6 EIA was used as a second-tier test, samples yielding C6 index values ≥3.0 could be considered positive. The identified cut-off values had also a high sensitivity to identify seropositivity among definite LB patients. The identified cut-off values refine the role of C6 EIA in LB serology. Importantly, the use of C6 EIA leads to a reduction in the number of samples that need to be analysed using an IB, thus also reducing the costs. Two alternative workflows for LB serology including the C6 EIA are suggested.


Subject(s)
Immunoenzyme Techniques/methods , Lyme Disease/blood , Lyme Disease/diagnosis , Peptides , Serologic Tests/methods , Adolescent , Adult , Aged , Aged, 80 and over , Antibodies, Bacterial/blood , Antibodies, Bacterial/isolation & purification , Bacteriological Techniques/methods , Borrelia burgdorferi/isolation & purification , Child , Child, Preschool , Female , Finland , Humans , Immunoblotting , Immunoglobulin G/blood , Immunoglobulin M/blood , Infant , Infant, Newborn , Male , Middle Aged , Retrospective Studies , Young Adult
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