Co-infections with Borrelia species, Anaplasma phagocytophilum and Babesia spp. in patients with tick-borne encephalitis.

The purpose of this study was evaluation of the prevalence of co-infection with Borrelia species, A. phagocytophilum and Babesia spp. in patients with tick-borne encephalitis (TBE). At total of 110 patients with TBE were included in the study. Serological tests for tick-borne encephalitis virus (TBEV), PCR for Borrelia species, Anaplasma phagocytophilum and Babesia spp., blood smears for A. phagocytophilum and Babesia spp. and BLAST analysis for Babesia spp. were performed. Results showed a significant majority of patients co-infected with Borrelia species (30/110; 27 %), much less with A. phagocytophilum (12/110; 10.9 %) and with Babesia spp. (1/110; 0.9 %). The BLAST analysis of the 18S rDNA sequence obtained with the Babesia spp. specific primers indicated that the patient was infected with Babesia microti. Triple co-infections (TBEV-Borrelia species- A. phagocytophilum) were observed in three (3/110; 2.7 %) patients. Conclusions were such that differential diagnosis in patients after the tick bite, presenting with acute symptoms, should include not only TBE and Lyme disease, but also other diseases transmitted by ticks. In patients with low parasitemia in suspicion of Babesia spp. infection PCR seems to be a more sensitive method than blood smear. Co-infection with various tick-borne pathogens must be always considered, especially in endemic regions.

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 Bialystok, 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.

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. METHODS: Clinical and epidemiological data of 687 patients hospitalized between 1993 and 2008 at the Department of Infectious Diseases and Neuroinfections with the diagnosis of TBE were analysed. RESULTS: In the case of 59 patients (9.5%), the disease was job related (forestry workers, farmers). In the examined group, TBE presented with meningitis in 282 cases (41%), with meningoencephalitis in 353 cases (51.3%) and with meningoencephalomyelitis in 52 cases (7.6%). The most common neurological abnormalities were ataxia in 88 cases (14.17%) and pareses in 53 cases (8.53%). Four patients (0.6%) died, 144 patients (23.2%) were discharged with neurological sequelae of TBE. Two hundred and seventy-two patients (43.8%) required further psychiatric treatment. At least 38 patients (6.1%) developed long-term sequelae and required further hospitalizations. Dexamethasone in the dosage of 6-32 mg was administered in 407 patients for 1-64 days. CONCLUSIONS: The diagnosis of TBE sometimes is difficult as the disease symptoms may be non-characteristic. Therefore, a detailed anamnesis is very important in the process of TBE diagnosis and may alone justify lumbar puncture conduction. Despite usually mild course of the disease, patients may develop neurological and psychiatrical sequelae.

Expression of Fas receptor on human T lymphocytes under stimulation with Borrelia burgdorferi sensu lato - preliminary results.

PURPOSE: Apoptosis of activated T lymphocytes is essential to immunoregulation and its abnormalities have been observed in immune system disorders and persistent infections. To asses Borrelia burgdorferi influence on the susceptibility of T lymphocytes to apoptosis, we have measured expression of the Fas death receptor on these cells after incubation with live B. burgdorferi. MATERIAL AND METHODS: Peripheral blood mononuclear cells from 23 LD patients (18 with Lyme arthritis, 5 with neuroborreliosis) and 13 healthy controls (C) were incubated for 48 hours with and without live B. burgdorferi spirochetes: B. afzelii, B. garinii or B. burgdorferi sensu stricto. After incubation, Fas expression on CD3+ cells was measured cytometrically with FITC-labeled monoclonal antibody. RESULTS: Median fraction of Fas-expressing T lymphocytes increased under incubation with B. burgdorferi, with more cells expressing Fas after incubation with B. burgdorferi sensu stricto than with B. garinii. There was a tendency for a higher expression of Fas on T lymphocytes from LD patients then from controls, both in unstimulated and B. burgdorferi-stimulated cultures, but it did not reach a level of statistical significance. CONCLUSIONS: B. burgdorferi seems to increase Fas expression on CD3+ T lymphocytes, which may render these cells more susceptible to apoptosis. This effect is stronger for B. burgdorferi s.s. than for B. garinii genospecies.

Concentration of TGF-beta1 in the supernatant of peripheral blood mononuclear cells cultures from patients with early disseminated and chronic lyme borreliosis.

PURPOSE: The aberrant inflammatory response is probably involved in the pathogenesis of chronic Lyme borreliosis, including chronic Lyme arthritis and neuroborreliosis. Transforming growth factor-beta 1 (TGF-beta1) is an important anti-inflammatory and immunomodulatory cytokine and its deficient synthesis is linked to exaggerated inflammation and immune response. MATERIAL AND METHODS: Peripheral blood mononuclear cells (PBMC) from 25 patients with Lyme borreliosis and 6 controls were incubated for 7 days with suspension of Borrelia afzeli, B. garinii and B. burgdorferi sensu stricto spirochetes. TGF-beta1 concentration in culture supernatants was measured with ELISA. Results were analyzed according to disease duration (group I--chronic borreliosis, n=20; group II--early borreliosis, n=5) and clinical form (LA--arthritis, NB--neuroborreliosis). RESULTS: TGF-beta1 concentration was increased in supernatants of PBMC cultures of patients with early neuroborreliosis, in comparison with chronic borreliosis and controls. In chronic, but not in early borreliosis, there was a tendency for decrease of TGF-beta1 synthesis under stimulation with B. burgdorferi spirochetes. CONCLUSIONS: Impaired synthesis of TGF-beta1 by mononuclear cells seems to be present in patients with chronic forms of Lyme borreliosis when compared to those with early stage of the disease. It may be a factor contributing to the persistence of inadequate inflammatory response in patients in whom chronic form of the disease develops.

Elevated concentration of the chemokine CCL3 (MIP-1alpha) in cerebrospinal fluid and serum of patients with tick borne encephalitis.

PURPOSE: Chemokines, including a chemoattractant for mononuclear cells CCL3 (MIP-1alpha), are responsible for attracting leukocytes into central nervous system (CNS) and cerebrospinal fluid (CSF) in meningitis and encephalomeningitis. We investigated the possibility of the involvement of CCL3 in tickborne encephalitis (TBE) pathogenesis. MATERIAL AND METHODS: We studied 26 patients with TBE; 13 with meningitis (group I) and 13 with encephalomeningits (group II). Control group included 11 patients without infectious disease of the CNS. CCL3 concentration was measured by ELISA in serum and CSF on admission (examination 1) and after 2 weeks (examination 2) in TBE patients and once in controls. RESULTS: In all control samples CCL3 concentration was below detection limit. In TBE, CCL3 serum concentration was: in group I--10.1 +/- 4.1 (mean +/- SD, ng/ml) in examination 1 and 12.4 +/- 4.8 in examination 2, and in group II--12.5 +/- 3.9 and 13.5 +/- 4.8, respectively. In CSF, CCL3 was detected: in group I in 5 patients in examination 1 (178 +/- 236 pg/ml) and 11 in examination 2 (457 +/- 215), in group II--in 8 (357 +/- 311) and 7 patients (326 +/- 330), respectively. There were no differences between group I and II. The comparison of CCL3 concentration gradient with albumin gradient between serum and CSF supported the possibility of intrathecal synthesis of CCL3. CONCLUSIONS: 1) Synthesis of CCL3, perhaps including intrathecal synthesis, is increased in TBE. 2) CCL3 concentration was much lower in CSF than in serum of the TBE patients, which argues against its significant role as chemoattractant in this condition.

Concentration of interferon-inducible T cell chemoattractant and monocyte chemotactic protein-1 in serum and cerebrospinal fluid of patients with Lyme borreliosis.

PURPOSE: Chronic inflammation in Lyme borreliosis may be sustained by aberrant inflammatory response, characterized by Th1 lymphocyte predominance, which in turn may be determined by chemokines synthesized in inflammatory focus. The aim of the study was to evaluate synthesis of chemokines: interferon-induced T cell chemoattractant (I-TAC--chemoattractant for Th1 lymphocytes), and monocyte chemotactic protein (MCP-1) in Lyme borreliosis. MATERIAL AND METHODS: Study group consisted of 13 patients with erythema migrans, 10 with Lyme arthritis and 6 with neuroborreliosis. Serum, as well as cerebrospinal fluid (CSF) in neuroborreliosis, was obtained before (examination 1) and during (examination 2) antibiotic treatment. Control serum was obtained from 8 healthy volunteers and control csf from 8 patients in whom meningitis and neuroborreliosis was excluded after diagnostic lumbar puncture. The samples were assayed for MCP-1 and I-TAC by ELISA. RESULTS: Serum mean I-TAC concentration in examination 1 was 73.0 pg/ml in erythema migrans, 78.9 pg/ml in Lyme arthritis and 87.3 pg/ml in neuroborreliosis (29.9 pg/ml in controls, difference significant for neuroborreliosis) and did not change significantly in examination 2. MCP-1 serum concentration was significantly increased to 497.5 pg/ml in neuroborreliosis in examination 2. I-TAC concentration in csf remained low, while MCP-1 concentration in examination 1 was increased to 589.1 pg/ml, significantly higher than simultaneously in serum. CONCLUSIONS: I-TAC synthesis is increased in Lyme borreliosis and may be a factor favoring predominance of Th1 lymphocyte subset. MCP-1 creates chemotactic gradient towards central nervous system and may contribute to csf pleocytosis in neuroborreliosis.

Concentrations of macrophage inflammatory proteins MIP-1alpha and MIP-1beta 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 1alpha and 1beta (MIP-1alpha, -1beta) 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 1beta (Il-1beta) 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-1alpha 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-1beta, 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-1alpha and 1beta 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-1alpha and 1beta) is increased in Lyme borreliosis and, at least in the early stages of the disease, is related to the synthesis of Il-1beta. 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.

[Chemokines in meningitis of different etiologies]. / Chemokiny w zapaleniach opon mózgowo-rdzeniowych o róznej etiologii.

Chemokines constitute a group of cytokines with strong chemotactic activity towards different populations of leukocytes, playing significant role in the pathogenesis of inflammatory responses. The chemokines of the alpha subfamily act mainly on neutrophiles, while beta subfamily chemokines attract primarily monocytes and lymphocytes. Research conducted within the last few years suggests chemokines to be the main factors responsible for the attraction of leukocytes to the cerebrospinal fluid (CSF) in the course of both bacterial and viral meningitis. In cerebrospinal fluid from patients with meningitis of different etiologies significant concentrations of both alpha and beta chemokines were observed, which tended to decrease after the introduction of the treatment, with the relationship to the clinical improvement. It was also confirmed in in vitro experiments that the chemotactic properties of the inflammatory CSF mainly depend on the presence of chemokines. The most important chemokines in the pathophysiology of the meningitis in humans are probably interleukin 8 (IL-8), monocyte chemoattractant protein (MCP-1) and INF-gamma inducible protein (IP-10). They seem to be responsible for the attraction to the cns of, respectively, neutrophiles, monocytes and activated T lymphocytes. Differences between the chemokine profiles observed in meningitides of different etiologies are to some degree relevant to coexisting differences in type and extent of pleocytosis. In future, measurements of concentrations of certain chemokines may become of some importance in diagnostics of meningitis and monitoring its clinical course.

[Epidemiologic aspect of lyme borreliosis among the inhabitants of Podlasie Province]. / Wybrane aspekty epidemiologiczne boreliozy z Lyme wsród mieszkanców województwa podlaskiego.

UNLABELLED: The aim of this study was to evaluate Lyme borreliosis morbidity among the inhabitants of Podlasie Province in the years of 1996-2000. MATERIAL AND METHODS: The new cases of Lyme borreliosis reported in the years of 1996-2000 in Podlasie Province were analyzed. The analysis included the data from Mz-56 and Mz-57 forms and the information from epidemiologic investigations sent to Regional Sanitary and Epidemiologic Station in Bialystok. In 1996-2000, screening examinations were carried out in the same group of 358 forestry workers including 44 (12.29%) females and 314 (87.71%) males aged from 21 to 64 (x = 41.2) years. Immunoenzymatic test of ELISA Borrelia recombinant IgM and IgG by Biomedica firm (Austria) were used to detect B. burgdorferi antibodies. RESULTS: In 1996-2000, 4933 of borreliosis cases were registered in Poland including 1377 (27.91%) in Podlasie Province. The morbidity rate in Podlasie province ranged from 15.05% in 1996 year to 21.29% in 2000 year of the whole country morbidity. At the same time, the incidence rate in Podlasie Province ranged from 9.09 in 1996 to 32.2 in 2000 year and was 6.72-fold higher than the incidence rate in the whole country. In Eastern and Central region of the province, 80.54% of cases were registered whereas, 14.09% by the Lakeside of Augustów and Suwalki only 5.37% in the western region of the province. It was proved that the morbidity increased in proportion to the age of patients (41.39% of patients were at the age of 30-49) and it decreased only above the age of 60. Thus, Lyme borreliosis affects mainly people at working age. The increase in B. burgdorferi antibody detectability was noticed in the population of forestry workers; it was detected in 38.55% in 1995 and in 2000 in 43.56% of the examined. Interestingly, 81 people infected with B.b in 1995-2000 included 74 men and only 7 women, supporting the statement that B.b infection affects mainly working professionals and the risk increases with the practice. CONCLUSIONS: Our studies indicate that Lyme borreliosis is a serious health problem among the inhabitants of Podlasie Province which is an endemic area for Ixodes ricinus ticks infected with B. burgdorferi spirochete.

[Acrodermatitis chronica atrophicans in the course of old Lyme disease]. / Przewlekle zanikowe zapalenie skóry konczyn w póznej boreliozie.

Lyme borreliosis is chronic spirochetal infection with phasic course. Lyme disease can be recognised as early or late stage of disease. A case report of patient with acrodermatitis chronica atrophicans in the course of chronic (late) Lyme borreliosis is presented. As it is described, the lack of uncognition of infection of B. burgdorferi and typical signs of erythema migrans as well as a lack of proper therapy in this time may be a reason of passing into chronic phase of the disease like ACA and Lyme arthritis.

[Poisoning with botulinum neurotoxin--diagnostic difficulties]. / Zatrucie toksyna botulinowa--trudnosci diagnostyczne.

Intoxication with botulinum neurotoxin, which occurs frequently in Poland, may cause serious diagnostic difficulties. As no assays for laboratory detection of botulinum toxin are available and the biological test on mice requires time before results are obtained, diagnosis must be based on clinical findings and patient's epidemiological history. Quick diagnosis and early administration of therapy with equine antitoxin is essential for patient's recovery. The effectiveness of antitoxin therapy is considered to be significantly reduced if the treatment is not started shortly after the onset of the disease. We describe a case of a patient in whom, because of diagnostic difficulties, antitoxin therapy was introduced with much delay. However, it proved highly efficient and lead to gradual recovery.

Analysis of some peripheral blood lymphocyte subsets in relation to Borrelia burgdorferi antibodies in patients with Lyme disease.

The aim of our study was to evaluate the changes in T (CD3), B(CD19) lymphocytes, CD4 and CD8 subsets, activated CD3+ HLA-DR+ lymphocytes, lymphocytes with receptor for IL-2 (CD3+CD25), NK cells as well as the rate of CD4/CD8 in 30 patients with recognized Lyme disease, before and after antibiotic therapy. Patients were divided into the following groups: Group I n = 9--without detected specific antibodies against B. burgdorferi and with clinical recognition of EM(erythema migrans). Group II n = 10--with increased IgM production, with a clinical form of Lyme arthritis and neuroborreliosis. Group III n = 11--with increased IgG anti B. burgdorferi production, with clinical recognition of Lyme arthritis. The results were compared with the results obtained in the control group consisting of 90 healthy people. The measurements were performed in the flow cytometer COULTER EPI XL with Becton Dickinson antibodies. The antibodies against B. burgdorferi were detected by means of ELISA method using Dako, Biomedica and Biocom kits. The statistic analysis was performed with AnStat Program. The changes in lymphocyte subsets were characterised by the decrease in the percentage of CD4, CD8, NK and CD3+HLA-DR+ lymphocytes in peripheral blood before treatment with later tendency to increase. The results show that the lag phase of antibodies production coincides with high activity of lymphocytes (group I). The significant level of antibodies in IgM class induced by the collaboration of T and B cells was found in the group II. In the group III, in which antibodies in IgG class predominated, the changes in lymphocyte subsets were less intensive. The results of our investigations indicate that the immune response in Lyme disease develops mainly with the participation of both cellular and humoral response which is involved in both the defense against and the pathogenesis of the disease.

[Graves-Basedow's disease: effect of treatment methods for hyperthyroidism on the course of ophthalmopathy]. / Choroba Gravesa-Basedowa: wplyw metody leczenia nadczynnosci tarczycy na przebieg oftalmopatii.

Efficient and permanent restoration of euthyreosis is an important factor in the therapy of Graves' ophthalmopathy (GO). The choice for a treatment method of thyreotoxicosis (thyreostatic drugs, radioiodine, strumectomy) may influence the natural course of the coexistent ophthalmopathy. Some forms of therapy seem to facilitate the withdrawal of the ophthalmic symptoms, while the others may worsen or even increase the risk of the reveal of GO. This paper includes the discussion of the influence of a chosen treatment of hyperthyreoidism on the course of GO.

[The prevalence of autoimmunological diseases in families of children with type I diabetes]. / Wystepowanie chorób o podlozu autoimmunoagresji w rodzinach dzieci chorych na cukrzyce typu I.

BACKGROUND: type I diabetes mellitus is considered as an autoimmune disease and is often associated with other diseases of that etiology. The genetic susceptibility to autoimmune disorders causes type I diabetes to occur more frequently in relatives of the diabetic patients. OBJECTIVE: To evaluate the prevalence of type I diabetes and other autoimmune diseases in families of the children with type I diabetes. MATERIAL AND METHODS: The prevalence of type I diabetes mellitus and other autoimmune endocrinopathies was evaluated in I, II and III degree relatives of 155 children with type I diabetes mellitus and 90 control children. RESULTS: It was observed that: 1) diabetes mellitus occurred more often in relatives of diabetic children (in 22 families - 14.2%) in comparison with the control group (in 2 families - 2.2%); 2) other autoimmune diseases occurred frequently in families of diabetic children and they affected 2 or more members of one family more often than in the control group (18 families vs 4); 3) rheumatoid arthritis occurred more frequently in families of diabetic children. CONCLUSIONS: The familial prevalence of type I diabetes mellitus and the tendency to more frequent prevalence of other autoimmune diseases in families of diabetic children was confirmed.