Grygorczuk S

Tick‐borne encephalitis in Poland in years 1993–2008 – epidemiology and clinical presentation. A retrospective study of 687 patients

Background and purpose:  Tick‐borne encephalitis (TBE) is an emerging disease in Europe as in Poland, especially in north‐eastern part of the country. The aim of the study was to characterize the epidemiology and clinical features of TBE in this region.

Evaluation of CXCL10, CXCL11, CXCL12 and CXCL13 chemokines in serum and cerebrospinal fluid in patients with tick borne encephalitis (TBE)

PURPOSE The aim of the study was to assess the concentration of chemokines: CXCL10, XCL11, CXCL12, CXCL13 in serum and cerebrospinal fluid (CSF) in patients with tick-borne encephalitis (TBE) before and after treatment. We evaluated also the usefulness of these molecules in diagnosis and monitoring of inflammation in TBE. METHODS Twenty three patients hospitalized in The Department of Infectious Diseases and Neuroinfections of Medical University in Białystok, Poland were included in the study. Patients were divided into 2 groups: TBE group-patients with confirmed TBE and control group (CG): patients with excluded TBE and other inflammatory diseases of CNS. Concentration of CXCL10/IP-10, CXCL11/I-TAC, CXCL12/SDF-1α, CXCL13/BLC/BCA-1 in serum and CSF were measured with ELISA kits (R&D Systems, USA) according to the protocols. RESULTS The analysis of chemokines concentration in TBE patients before treatment and control group using ROC showed that serum CXCL10 and CXCL13 and CSF CXCL10, CXCL11, CXCL12 and CXCL13 differentiate both groups (p<0.05). The analysis of CXCL10, CXCL11, CXCL12 and CXCL13 before and after treatment showed that CXCL10 and CXCL11 in CSF and CXCL13 in serum differentiates both groups with p<0.05. CONCLUSIONS Concentration of CSF CXCL10, CXCL11, CXCL12, CXCL13 and serum CXCL10, CXCL13 may be good biomarkers of CNS inflammation caused by TBEV. Moreover concentration of CXCL10 in CSF and CXCL13 in serum may be used as indicators of patients recovery.

Concentrations of Macrophage Inflammatory Proteins MIP-1α and MIP-1β and Interleukin 8 (Il-8) in Lyme Borreliosis

Background:Components of the spirochete Borrelia burgdorferi sensu lato (B. burgdorferi s.l.) do not have chemotactic activity. However, B. burgdorferi s.l. causes a chemotactic response, probably by stimulating synthesis of cytokines of the chemokine family by host cells. Our aim was to confirm that the synthesis of chemokines is increased in Lyme borreliosis and that they may account for leukocyte migration, thus being involved in inflammatory response.Materials and Methods:We measured concentrations of chemokines: interleukin 8 (Il-8) and macrophage inflammatory protein 1α and 1β (MIP-1α, -1β) in serum of 20 patients with erythema migrans (early localized infection, group I), of 19 patients with Lyme arthritis (chronic infection, group II), and in serum and cerebrospinal fluid (CSF) of 20 patients with neuroborreliosis (early disseminated infection, group III), before and after 2 weeks of antibiotic therapy (examinations 1 and 2), as well as in the sera of 12 healthy blood donors and CSF of ten patients in whom Lyme borreliosis and meningitis were excluded (control group). Interleukin 1β (Il-1β) level in serum and CSF and pleocytosis of CSF were assessed simultaneously.Results:The mean concentrations of all studied chemokines in serum were significantly elevated in all study groups in examination 1 and decreased in examination 2. The concentration of Il-8 in serum was higher in group I and the concentration of MIP-1α in group III was higher in comparison with group II. Serum concentrations of all chemokines in group I and III correlated with the concentration of Il-1β, while in group II this correlation appeared only for Il-8 in examination 2. Concentrations of all chemokines in CSF were significantly increased, but as for MIP-1α and 1β they remained lower than in serum. The concentration of Il-8 in CSF was variable and reached values several fold higher than in the serum in some patients. There was no correlation between chemokine concentrations and CSF pleocytosis.Conclusion:The synthesis of chemokines (Il-8, MIP-1α and 1β) is increased in Lyme borreliosis and, at least in the early stages of the disease, is related to the synthesis of Il-1β. Chemokine concentrations depend on the clinical form of Lyme borreliosis, with a tendency for higher values in early infection (erythema migrans and neuroborreliosis). Of the chemokines studied, Il-8 created a chemotactic gradient towards the inflammation site, and thus might be responsible for leukocyte migration.

Activity of lysosomal exoglycosidases in serum and synovial fluid in patients with chronic Lyme and rheumatoid arthritis

Lysosomal exoglycosidases participate in the destruction of the articular cartilage by cleaving glycoside bonds in glycoproteins and proteoglycans. The aim of the study was to determine the activity of exoglycosidases: hexosaminidase, β-glucuronidase, β-galactosidase, α-mannosidase and α-fucosidase in serum and synovial fluid of patients with Lyme and rheumatoid arthritis. The study group consisted of 10 patients with chronic Lyme arthritis (age 18 – 74 y), 13 with rheumatoid arthritis (age 32 – 70 y) and 10 with juvenile idiopathic arthritis (age 8 – 17 y). The control group consisted of 9 healthy volunteers (age 24 – 62 y). The activity of the exoglycosidases was determined with the p-nitrophenyl derivatives of sugars as substrates. A significant increase of the activity of all the exoglycosidases in serum and in synovial fluid of the patients with different forms of arthritis was found. The ratio of synovial fluid/serum activity of exoglycosidases was above 2.0 in LA but not in JIA and RA patients. As the main source of exoglycosidases in the joint is the synovial membrane, this result supports the appropriateness of therapeutic synovectomy in chronic Lyme arthritis with knee effusion. The serum activity of hexosaminidase may be used in monitoring the course of Lyme arthritis and the efficiency of treatment.

Evaluation of CXCL8, CXCL10, CXCL11, CXCL12 and CXCL13 in serum and cerebrospinal fluid of patients with neuroborreliosis.

PURPOSE Knowledge of the role of chemokines in the inflammation during neuroborreliosis (NB) is limited. We evaluated the pre- and post-treatment concentration of CXCL8, CXCL10, CXCL11, CXCL12, and CXCL13 in serum (s) and cerebrospinal fluid (csf) in patients with NB. RESULTS There was a statistically significant increase in pre-treatment s CXCL8, CXCL10, CXCL11, CXCL12, CXCL13 and csf CXCL8, CXCL11, CXCL12, CXCL13 in patients with early form of NB. CXCL8, CXCL11, CXCL12 and CXCL13 increase was the highest in csf. After treatment, a significant decrease in csf chemokine levels (except CXCL10) and s levels (except CXCL11) was observed. CONCLUSIONS CXCL8, CXCL10, CXCL11, CXCL12, CXCL13 are involved in the pathomechanism of NB but their role is different in s and csf. CXCL13 seems to be a good biomarker for NB. In early NB, it may facilitate the diagnosis and monitoring of therapy. However tick-borne encephalitis needs to be excluded as it also increases chemokine concentration. Decrease in all examined chemokines in s and csf after treatment suggests that chemokines may be useful in monitoring response to NB therapy.

Increased concentration of interferon lambda-3, interferon beta and interleukin-10 in the cerebrospinal fluid of patients with tick-borne encephalitis

Tick-borne encephalitis (TBE) has a wide clinical spectrum, from asymptomatic to severe encephalitis, and host-dependent factors determining the outcome remain elusive. We have measured concentrations of pro-inflammatory/Th1 interferon-γ (IFNγ), immunomodulatory/Th2 interleukin-10 (IL-10), anti-viral type I (IFNβ) and type III (IFNλ3) interferons in cerebrospinal fluid (csf) and serum of 18 TBE patients, simultaneously genotyped for polymorphisms associated with the expression of genes IFNL3 (coding IFNλ3), IL10, CD209 and CCR5. IL-10, IFNβ and IFNλ3 were up-regulated in csf, with IFNλ3 level higher in patients with the milder clinical presentation (meningitis) than in meningoencephalitis. There was an increased serum IFNβ and a tendency for increased serum IL-10 in meningitis patients. Genotype in rs12979860 locus upstream of IFNL3 was associated with IFNλ3 expression and in rs287886 (CD209) - IL-10 expression. IL-10, IFNβ and IFNλ3 are expressed and play a protective role in TBE and their expression in TBE patients is associated with genetic polymorphisms.

Anti-Babesia microti antibodies in foresters highly exposed to tick bites in Poland.

Abstract Background: Human babesiosis caused by Babesia microti and Babesia divergens parasites is an emerging tick-borne disease worldwide. The prevalence of infection and frequency of the disease caused by B. microti in Europe is not well known. The aim of our study was to evaluate the frequency of anti-B. microti antibodies in the serum of forest employees (a population highly exposed to tick bites) from 2 different regions of Poland. Methods: We studied 114 foresters from 2 separate forest inspectorates in north-eastern and central Poland. Direct immunofluorescence assays (Babesia microti IgM and IgG IFA kits) were used to detect serum IgM and IgG anti-B. microti antibodies. Simultaneously, anti-B. burgdorferi antibodies were detected using an enzyme-linked immunosorbent assay kit, and positive cases were confirmed with immunoblot. Results: Anti-B. microti IgG antibodies were detected in 5 foresters (4.4%), all from the forest inspectorate in Białowieża in the northeast of the country. All persons with anti-B. microti antibodies were also IgG-seropositive for B. burgdorferi. Conclusions: Our results suggest that unrecognized infections with B. microti occur in the Polish population and should be considered in the differential diagnosis of a febrile illness occurring after exposure to ticks, particularly in patients from endemic regions.

The expression of the chemokine receptor CCR5 in tick-borne encephalitis

BackgroundChemokine receptor 5 (CCR5) is hypothesized to drive the lymphocyte migration to central nervous system in flavivirus encephalitis, and the non-functional CCR5Δ32 genetic variant was identified as a risk factor of a West Nile virus infection and of tick-borne encephalitis (TBE). We have attempted to investigate how CCR5 expression corresponds to the clinical course and severity of TBE.MethodsWe have repeatedly studied CCR5 expression in 76 patients during encephalitic and convalescent TBE phase, analyzing its association with clinical features, cerebrospinal fluid (csf) pleocytosis, and concentrations of CCR5 ligands (chemokines CCL3, CCL4, and CCL5) and CCR5 genotype. Fifteen patients with neuroborreliosis, 7 with aseptic meningitis, 17 in whom meningitis/encephalitis had been excluded, and 18 healthy blood donors were studied as controls. Expression of CCR5 was measured cytometrically in blood and csf-activated Th lymphocytes (CD3+CD4+CD45RO+). Concentrations of chemokines in serum and csf were measured immunoenzymatically, and CCR5Δ32 was detected with sequence-specific primers. Data were analyzed with non-parametric tests, and p < 0.05 was considered significant.ResultsThe blood expression of CCR5 did neither differ between the groups nor change in the course of TBE. The CCR5 expression in the inflammatory csf was several-fold increased in comparison with blood but lower in TBE than in neuroborreliosis. The csf concentration of CCL5 was increased in TBE, the highest in the most severe presentation (meningoencephalomyelitis) and correlated with pleocytosis. The CCR5Δ32/wt genotype present in 7 TBE patients was associated with a decreased CCR5 expression, but enrichment of csf Th population in CCR5-positive cells and the intrathecal inflammatory response were preserved, without a compensatory increase of CCL5 expression.ConclusionsWe infer CCR5 and CCL5 participate in the response to TBE virus, as well as to other neurotropic pathogens. The intrathecal response to TBE is not hampered in the bearers of a single copy of CCR5Δ32 allele, suggesting that the association of CCR5Δ32 with TBE may be mediated in the periphery at the earlier stage of the infection. Otherwise, a variability of the CCR5 expression in the peripheral blood lymphocytes seems not to be associated with a variable susceptibility to TBE.

ssICAM-1, IL-21 and IL-23 in patients with tick borne encephalitis and neuroborreliosis

Abstract Objective There have been few reports on the role of Intercellular Adhesion Molecule 1 (ICAM-1), but not interleukin-21 (IL-21) and interleukin-23 (IL-23) in tick-borne encephalitis (TBE) and neuroborreliosis (NB). We postulate that these two interleukins may participate in the early phase of TBE and NB. The aim of the study was to measure serum and cerebrospinal fluid (CSF) concentration of ICAM-1, IL-21 and IL-23 in patients with TBE and NB before treatment and to assess their usefulness in the diagnosis and monitoring of inflammatory process in TBE and NB. Methods Forty-three patients hospitalized in The Department of Infectious Diseases and Neuroinfections of Medical University in Bialystok, Poland, were included in the study. Patients were divided into three groups: TBE, NB and CG. Pre-treatment blood and CSF samples were obtained from all patients. ELISA kits (DRG Instruments, Germany) were used to measure the concentration of IL-21, IL-23 and sICAM-1. Results Significant differences between TBE/CG and NB/CG concentration of sICAM-1 were found only in the CSF. CSF IL-21 levels in NB were lower than in TBE. In TBE, a strong negative correlation between CSF concentration of IL-21 and IL-23 and monocyte count in CSF was observed. Negative correlation between IL-21 in CSF and neutrophil count was also noted. Serum IL-23 correlated positively with leukocytes and platelet count in serum. In NB, a strong positive correlation between serum IL-21 and platelet count and negative correlation between IL-21 in serum and CSF with pleocytosis was observed. Conclusions Increased sICAM-1 concentration in TBE and NB may be a proof of brain–blood barrier disturbances in the early phase of these diseases. IL-21 and IL-23 do not appear to play an important role in the pathogenesis of the early stages of TBE and NB.

Tick-borne encephalitis in north-eastern Poland in 1997-2001: a retrospective study.

The aim of this study was to characterize the epidemiology and clinical features of tick-borne encephalitis in north-eastern Poland. Clinical and epidemiological data were analysed of patients hospitalized with the diagnosis in the Department of the Infectious Diseases and Neuroinfections of the Medical University in Bialystok in 1997-2001. Tick-borne encephalitis was diagnosed in 152 patients: 51 (34%) presented with meningitis, 89 (59%) with meningoencephalitis and 12 (8%) with meningoencephalomyelitis. Headache (84%) and fever (81%) were the most common symptoms. Meningeal signs were present in 137 patients (90%). Most common neurological abnormalities were: Oppenheim and Babinski signs (74 patients, 49%), ataxia (37, 24%), impaired consciousness (37, 24%) and pareses (16, 10%). Of patients examined, 146 (96%) had raised pleiocytosis, frequently accompanied by high cerebrospinal fluid protein concentration (90%), raised erythrocyte sedimentation rate (65%), peripheral blood leucocytosis (26%) and increased aminotransferase activity (16%). There was only 1 forest worker among the patients. Tick-borne encephalitis remains common in north-eastern Poland but, possibly because of effective vaccination, it has virtually disappeared among forest employees. The diagnosis appears difficult in some cases, as meningeal signs may not be present and laboratory findings may not be suggestive of a viral infection.