Differences in Genotype, Clinical Features, and Inflammatory Potential of Borrelia burgdorferi sensu stricto Strains from Europe and the United States.

Borrelia burgdorferi sensu stricto isolates from patients with erythema migrans in Europe and the United States were compared by genotype, clinical features of infection, and inflammatory potential. Analysis of outer surface protein C and multilocus sequence typing showed that strains from these 2 regions represent distinct genotypes. Clinical features of infection with B. burgdorferi in Slovenia were similar to infection with B. afzelii or B. garinii, the other 2 Borrelia spp. that cause disease in Europe, whereas B. burgdorferi strains from the United States were associated with more severe disease. Moreover, B. burgdorferi strains from the United States induced peripheral blood mononuclear cells to secrete higher levels of cytokines and chemokines associated with innate and Th1-adaptive immune responses, whereas strains from Europe induced greater Th17-associated responses. Thus, strains of the same B. burgdorferi species from Europe and the United States represent distinct clonal lineages that vary in virulence and inflammatory potential.

Early production of IL-22 but not IL-17 by peripheral blood mononuclear cells exposed to live Borrelia burgdorferi: the role of monocytes and interleukin-1.

If insufficiently treated, Lyme borreliosis can evolve into an inflammatory disorder affecting skin, joints, and the CNS. Early innate immunity may determine host responses targeting infection. Thus, we sought to characterize the immediate cytokine storm associated with exposure of PBMC to moderate levels of live Borrelia burgdorferi. Since Th17 cytokines are connected to host defense against extracellular bacteria, we focused on interleukin (IL)-17 and IL-22. Here, we report that, despite induction of inflammatory cytokines including IL-23, IL-17 remained barely detectable in response to B. burgdorferi. In contrast, T cell-dependent expression of IL-22 became evident within 10 h of exposure to the spirochetes. This dichotomy was unrelated to interferon-γ but to a large part dependent on caspase-1 and IL-1 bioactivity derived from monocytes. In fact, IL-1ß as a single stimulus induced IL-22 but not IL-17. Neutrophils display antibacterial activity against B. burgdorferi, particularly when opsonized by antibodies. Since neutrophilic inflammation, indicative of IL-17 bioactivity, is scarcely observed in Erythema migrans, a manifestation of skin inflammation after infection, protective and antibacterial properties of IL-22 may close this gap and serve essential functions in the initial phase of spirochete infection.

Successful amplification of DNA specific for Finnish Borrelia burgdorferi isolates in erythema chronicum migrans but not in circumscribed scleroderma lesions.

Early diagnosis of Borrelia burgdorferi infection, hampered by the absence of detectable antibodies in most patients with erythema chronicum migrans is important to prevent late-stage neurologic, rheumatologic, and skin disorders. Furthermore, B. burgdorferi has been claimed to be the causative agent in localized scleroderma (morphea). We used PCR amplification to search for B. burgdorferi outer surface protein OspA-specific sequences in DNA obtained from lesional skin biopsies on Finnish patients with clinically suspect erythema chronicum migrans, lymphocytoma, morphea, or with diverse skin manifestations and persistent high antibodies to B. burgdorferi flagellar antigen. Seronegative patients with other skin lesions served as controls. The amplicons obtained with primers specific for B. burgdorferi type strain B31 ospA sequence did not hybridize to the corresponding probes, and thus the DNA amplified from a Finnish B. burgdorferi erythema chronicum migrans skin isolate was sequenced. This 98-nucleotide sequence of ospA (332-429) showed 11% to 14% nucleotide divergence compared with the North American type strain (B31), several European strains, and an East Siberian tick strain. The sequence was almost identical (99%) to a Swedish isolate from acrodermatitis chronica atrophicans. Using oligonucleotides specific for the Finnish strain, a positive polymerase chain reaction-based hybridization was obtained in six of seven untreated erythema chronicum migrans patients infected in Finland or in Estonia, and in the lymphocytoma patient. Only two of the erythema chronicum migrans patients had IgG or IgM antibodies to flagellin. However, all seven morphea lesions as well as the other lesions were polymerase chain reaction negative. Polymerase chain reaction-based hybridization of B. burgdorferi OspA gene from skin-derived DNA thus provides a sensitive and specific diagnostic tool. In conditions not unequivocally known to be caused by B. burgdorferi, like in morphea, this assay was negative. We also demonstrate that peri-Baltic B. burgdorferi isolates show homology in their OspA genes but differ from geographically more distant isolates.

[Cutaneous forms of Lyme borreliosis in children]. / Kozní formy lymeské borreliózy u detí.

Thirty-seven children with skin manifestations of Lyme borreliosis (31 with erythema chronicum migrans and six with lymphadenosis benigna cutis) were treated and followed up prospectivelly for 1-24 months (mean 7.58 months). The diagnosis was confirmed serologically by the finding of increased levels of antiborrelial antibodies assessed by the ELISA method, using antigen from Borrelia recurrentis, in a total of 87% patients with erythema chronicum migrans and in all patients with lymphadenosis benigna cutis. In nine patients the dermatoses were associated with general non-characteristic symptoms, after antibiotic treatment in five patients temporary fatigue, arthralgia and gastrointestinal complaints were observed. No extradermal organ complications were present. The authors discuss the clinical pictures of erythema chronicum migrans and lymphadenosis benigna cutis in children, mention diagnostic criteria and the recommended pattern of treatment.

[Role of interleukin-18, interleukin-1beta and its soluble receptor (sIL-1RII) in early and late Lyme borreliosis]. / Udzial interleukiny 18, interleukiny 1beta i jej rozpuszczalnego receptora (sIL-1RII) u chorych we wczesnej i póznej postaci boreliozy z Lyme.

Although borreliosis was first described as a separate entity more than 20 years ago its pathogenesis still remains unknown. In recent years the role of pro- and antiinflammatory cytokines in the pathogenesis of borreliosis has been discussed. The purpose of the present study was to evaluate the role of IL-1beta, IL-18 and sIL-1RII in the development of early and late stages of borreliosis. The study group consisted of 60 patients divided into 3 groups: patients with erythema migrans, Lyme arthritis and neuroborreliosis. In all groups serum levels of IL-1beta, IL-18 and sIL-1RII were determined and in the patients with neuroborreliosis additionally in cerebrospinal fluid (CSF). The levels of cytokines and sIL-1RII were measured before the start of treatment and after its termination. Before the treatment the levels of IL-1beta, IL-18 and sIL-1RII in serum and CSF were significantly higher in all studied groups compared with the control. After the treatment, despite the regression of the clinical symptoms and significant reduction of initially high levels of the cytokines and sIL-1RII, only the levels of IL-1beta in all patients and the serum level of IL-18 in the patients with neuroborreliosis were comparable with the values in the control group. It could suggest that the inflammatory process was not inhibited completely and confirms the role of IL-1beta, IL-18 and sIL-1RII in the pathogenesis of borreliosis.

Role of reactive oxygen species (ROS) in patients with erythema migrans, an early manifestation of Lyme borreliosis.

BACKGROUND: Lyme borreliosis is a tick-transmitted, chronic, zoogenous disease caused by Borrelia burgdorferi spirochete. The clinical picture of Lyme disease is characterized by the variety of tissue and organ involvement and differing severity of symptoms. One of the pathogenic symptoms of early Lyme disease is a skin lesion called erythema migrans. MATERIAL AND METHODS: The purpose of our research was to estimate the parameters of the antioxidant system and the concentration of lipid peroxidation products in the plasma of patients with erythema migrans (EM). The parameters measured included the activity levels of superoxide dismutase (SOD) according to Sykes, gluthatione reductase (GSSG-R) according to Mize and Langdon, glutathione peroxidase (GSH-Px) according to Paglia and Valentine; the concentrations of malondialdehyde (MDA) were examined by means of a Bioxytech LPO-586 kit. The total sulphydryl groups (-SH) according to Ellman and reduced glutathione (GSH) were measured using a Bioxytech GSH-400 test in plasma samples collected from 20 patients with EM aged from 19 to 50, taken before (examination 1) and after (examination 2) therapy with amoxycycline. The control group consisted of 8 healthy people. RESULTS: The results of our examinations prove that beta-lactamase antibiotic therapy brings non-enzymatic antioxidant parameters to control values, though the treatment causes no change in enzymatic antioxidant parameters, resulting in the further activation of free radicals. CONCLUSIONS: In patients with Erythema migrans, the decreased capability to reduce lipid superoxidants leads to maintaining a high concentration of membrane lipid peroxidation products.

Azithromycin versus doxycycline for treatment of erythema migrans: clinical and microbiological findings.

The effectiveness of azithromycin and doxycycline in the treatment of erythema migrans was compared in a prospective randomized trial. One hundred seven adult patients with typical erythema migrans, examined in the Lyme Borreliosis Outpatients' Clinic, University Department of Infectious Diseases in Ljubljana, were included in the study. Fifty-five patients received azithromycin (500 mg twice daily for the first day, followed by 500 mg once daily for four days) and 52 patients received doxycycline (100 mg twice daily for 14 days). The mean duration of skin lesions after the beginning of treatment was 7.5 +/- 5.9 days (median value 5, range 2-28 days) in the azithromycin group and 11.4 +/- 7.8 days (median value 9, range 2 days--8 weeks) in the doxycycline group (p < 0.05). Borrelia burgdorferi was isolated from erythema migrans in 28 patients before therapy: in 13 out of 52 in the doxycycline group and in 15 out of 55 in the azithromycin group. Three months after therapy, the culture was positive in four out of 13 patients treated with doxycycline and in one of the 15 patients who received azithromycin. A biopsy was repeated in all the patients with a positive isolation from the first skin specimen. During the first 12 months' follow-up, three patients treated with doxycycline but none in the azithromycin group developed major manifestations of Lyme borreliosis, while 15 doxycycline recipients and 10 azithromycin recipients developed minor consecutive manifestations.