Expression of Genes Associated with H Factor in Fibroblasts Infected with Borrelia Spirochaetes

Abstract

Infection with Borrelia spirochaetes leads to the launch of specific and non‐specific immunological response in humans. Activation of the complement system is one of the first defence mechanisms against penetrating pathogens. The aim of this study was to select genes related to the alternative pathway of the complement system [including complement factor H (CFH)], differentiating the type of infection in the system model, that is, a culture of normal human diploid fibroblasts infected with three different spirochaete genospecies: Borrelia afzelii, Borrelia garinii and Borrelia burgdorferii sensu stricto, by comparing the infected fibroblast culture with the control fibroblast one. With the use of oligonucleotide microarrays HGU‐133A, the differences in the expression of genes selected on the basis of a scientific database Affymetrix were studied by comparing transcriptomes from the four cultures of fibroblasts. In the result of infection of fibroblast cultivation with a specific Borrelia genospecies, a variable expression of certain CFH and complement system‐associated genes, specific for one genospecies only, B. afzelii– C1QBP, CD59, C2, CD46 and FHL1; B. garinii – C1S and CLU; Borrelia burgeforii– CFB, A2M and VSIG4, was observed. CFH differentiates infections induced by B. afzelii and B. garinii from infections induced by B. burgdorferii sensu stricto.

Fibroblast Protection ofBorrelia burgdorferifrom Doxycycline, Cefuroxime and Daptomycin Combination is Eliminated by Oregano or Carvacrol Essential Oil

“…When the confluence stage was reached, GFP-labeled B. burgdorferi at stationary phase of growth (SP) was centrifuged at 10,000 rpm for 10 min and resuspended in DMEM supplemented with 10% FBS. Then GFP B. burgdorferi was added to tissue culture plate wells containing 3T3 cells in a 1:10 ratio of fibroblasts against the bacteria or to wells without cells (31). Cells and B. burgdorferi were cocultured for 48 h. Then the cells were then washed twice with warm PBS, fresh DMEM medium containing cefuroxime, doxycycline and daptomycin at 5 μg/mL or essential oils at three different concentrations 0.1%, 0.05% and 0.02% were added and the cocultures were further incubated for 7 days before the effect of the treatment was evaluated by microscopy or subculture study (5).…”

The Impact of Three Genospecies of Borrelia on Expression of Genes Associated with Chemokines and Their Receptors in Normal Human Dermal Fibroblastsin Vitro

“…Migration of leukocytes from the extravascular space into tissues is a basic phenomenon deciding on maintenance of the immunological control. It requires a multi-stage, cascade adhesion with, among others, selectins and integrins, and presence of appropriate vascular ligands (8,9). Chemokines mediate in integrin activation and a strong adhesion ofcells to epithelium of small blood vessels, and their migration through vascular walls and further on within extravascular tissues.…”