Neutrophil chemotactic factors in synovial fluids of patients with Lyme disease

We examined synovial fluid samples from 14 patients with Lyme arthritis for the presence of neutrophil chemotactic factors. Thirteen of the synovial fluids stimulated chemotaxis of normal human neutrophils. The chemotactic activity was heat-sensitive and was not inhibited by antibody to C5a or antibody to interleukin-8, or by a competitive inhibitor of the chemotactic peptide f-Met-Leu-Phe. A culture supernatant of Borrelia burgdorferi also contained neutrophil chemoattractants. Chromatography demonstrated that the chemoattractants in the synovial fluid samples were different from those in the B burgdorferi culture supernatant. One of the major chemotactic factors in Lyme disease synovial fluid had a calculated molecular weight of 13,900. We conclude that a novel, host-derived chemoattractant is present in the synovial fluid of patients with Lyme disease.     

Prevalence of antibody to Borrelia burgdorferi in children with juvenile rheumatoid arthritis

Lyme arthritis and juvenile rheumatoid arthritis (JRA) share a number of clinical features. Our study was performed in order to determine the prevalence of antibody to Borrelia burgdorferi in 50 children with JRA who reside in a nonendemic area. Three patients were weakly reactive and one patient was reactive when tested using an enzyme immunoassay to detect serum antibody to B. burgdorferi. No patient, however, had definitive serologic evidence of B. burgdorferi infection by Western blot analysis. We conclude that the prevalence of antibody to B. burgdorferi is very low in children with JRA who reside in a nonendemic area.     

Seroreactivity to Borrelia burgdorferi antigens in early rheumatoid arthritis: a case-control study

This study investigates the seroreactivity to Borrelia burgdorferi antigens of patients suffering from rheumatoid arthritis (RA) for < 5 yr. Subjects were matched with controls for age, sex and area of residence in order to minimize the risk of differential exposure to B. burgdorferi. A total of 57 pairs were tested by immunofluorescence assay (IFA) and Western blotting. Only two RA patients showed positive IFA results, and another was positive by Western blot analysis. No significant difference in Lyme seropositivity was detected between cases and controls using either serological test. Furthermore, no significant difference in antibody response was observed to specific or non-specific proteins (e.g. Osp C, 93 kDa protein, flagellin, heat shock proteins). These results do not support the previous suggestions that B. burgdorferi may be involved in the aetiology of RA, or that RA favours the production of antibodies that cross-react with B. burgdorferi proteins.      

Immune responses to Borrelia burgdorferi in patients with reactive arthritis

In reactive arthritis (ReA), including Reiter's syndrome, a close relationship between chronic enteric and genitourinary infections and the clinical features of enthesitis has been described. In contrast, in Lyme arthritis, a distinct clinical entity, chronic infection with the tick-transmitted spirochete Borrelia burgdorferi has been associated with the disease. In a prospective study, 51 patients with ReA were tested for evidence of chlamydial and spirochetal infection. The presence of Chlamydia was determined by culture in 8 patients, and 7 additional patients had markedly elevated antibody titers. In 9 patients, antibodies specific to B burgdorferi were found. Purified peripheral blood T lymphocytes of all 9 patients proliferated specifically to stimulation with macrophages pre-pulsed with B burgdorferi antigens. Compared with other protein antigens, higher numbers of antigen-pulsed macrophages were necessary to activate B burgdorferi-specific T cells. Although antibody titers decreased in response to antibiotic treatment in 8 of 9 patients, second-line therapy with sulfasalazine or methotrexate was required to obtain clinical remission. These data suggest that chronic infection with B burgdorferi can cause ReA. In predisposed individuals, the arthritogenic immune response might be triggered by persisting infectious agents independent of their antigenic specificities.     

结节病病人血清抗布氏疏螺旋体抗体测定及其意义

用酶联免疫检测法（ＥＬＩＳＡ）测定４０例患者血清抗布氏疏螺旋体（ＢｏｒｒｅｌｉａＢｕｒｇｄｏｒｆｅｒｉ，简称ＢＢ）抗体水平，其中结节病２３例，非结节病１７例。结节病病人血清抗ＢＢ抗体阳性１８例，占７８．３％，非结节病病人阳性１３例，占７６．５％。初步认为结节病可能是一种自身免疫异常疾病，血清抗ＢＢ抗体可能是机体对某种不明抗原刺激所产生的一种自体抗体，与风湿病、结核病、肿瘤和莱姆病可能存在着交叉反应；但是否有另外一种可能：即ＢＢ抗原本身是一种混合物，不同疾病中的不同抗体与ＢＢ抗原中的不同组分起反应？因此，ＢＢ抗体不能作为诊断结节病的特异性指标。     

Reversal of Borrelia burgdorferiassociated dilated cardiomyopathy by antibiotic treatment?

Summary It is suggested thatBorrelia burgdorferi infection could be associated with dilated cardiomyopathy (IDC). Stanek et al. were able to cultivateBorrelia burgdorferi from myocardial biopsy tissue of a patient with longstanding dilated cardiomyopathy. Here we present a study in which we examined the effect of standard antibiotic treatment on the left ventricular ejection fraction (LVEF) in patients with dilated cardiomyopathy associated withBorrelia burgdorferi infection. In this study we assessed the serum (IgG, IgM Elisa) and history of 46 IDC patients with specific regard toBorrelia burgdorferi infection (mean LVEF 30.4±1.3%, measured by cardiac catheterization and echocardiography with the length-area-volume method). All 46 patients received standard treatment for dilated cardiomyopathy: ACE inhibitors, digitalis, and diuretics. Eleven (24%) patients showed positive serology and a history ofBorrelia burgdorferi infection; nine of these also had a typical history of tick bite and erythema chronicum migrans (ECM) and/or other organ involvement, and two had no recollection of tick bite or ECM but showed otherBorrelia burgdorferi-associated disorders (neuropathy, oligoarthritis). These 11 patients withBorrelia burgdorferi infection received standard antibiotic treatment with intravenous ceftriaxone 2 g bid for 14 days. Six (55%) recovered completely and showed a normal LVEF after 6 months, three (27%) improved their LVEF, and two (18%) did not improve at all. This amounts tonine (82%) patients with recovery/improvement in the Borrelia burgdorferi group. The 35 patients who did not show positive serology or a history ofBorrelia burgdorferi infection did not receive antibiotic treatment. In thisgroup without Borrelia burgdorferi infection 12 (26%), showed recovery/improvement following the standard treatment of dilated cardiomyopathy (see earlier). Our results indicate thatBorrelia burgdorferi infection could play a decisive role in the development of dilated cardiomyopathy, especially in a geographical region such as Graz, whereBorrelia burgdorferi is endemic. While we are aware of the small number ofBorrelia burgdorferi patients in this study, we nevertheless conclude that in a remarkable number of patients with signs ofBorrelia burgdorferi infection, dilated cardiomyopathy could be reversed and LVEF improved.     

No evidence to implicate Borrelia burgdorferi in the pathogenesis of dilated cardiomyopathy in the United Kingdom.

OBJECTIVE--To determine whether Borrelia burgdorferi is implicated in the pathogenesis of dilated cardiomyopathy in the United Kingdom. DESIGN--A controlled prospective study. Patients' notes were reviewed for evidence of Lyme disease and serum samples were tested by enzyme linked immunoadsorbent assay (ELISA) for antibodies to B burgdorferi. Samples with raised antibody concentrations were subsequently analysed by immunoblotting to determine their antibody binding specificity. SETTING--Tertiary referral centre. PATIENTS--97 consecutive patients with dilated cardiomyopathy diagnosed according to World Health Organisation criteria were studied. Serum samples were taken from two matched control groups. The first group (n = 38) was age, sex, and geographically matched. The second control group (n = 39) was environmentally matched and consisted of members of the patients' own households. MAIN OUTCOME MEASURES--Clinical evidence of Lyme disease. Presence of raised antibody concentrations to B burgdorferi. RESULTS--No patients had a previous illness compatible with Lyme disease. Analysis of the ELISA data showed eight of 97 patients with dilated cardiomyopathy (8.2%) and two of 77 controls (3.9%) had raised antibody concentrations. Immunoblot analysis, however, did not show binding patterns consistent with the presence of IgG specific for B burgdorferi in any of these samples. CONCLUSIONS--There was no clinical or serological evidence to implicate B burgdorferi in the pathogenesis of idiopathic dilated cardiomyopathy in the United Kingdom. In the absence of specific symptoms or likely exposure to B burgdorferi routine serological testing for Lyme disease in this group of patients is not recommended. Furthermore, raised antibodies to B burgdorferi are not diagnostic of active infection and ELISA results should be interpreted with caution unless specific B burgdorferi antibody bands have been found by immunoblot analysis.     

Study of the anti Borrelia burgdorferi antibody of hunters in Hokkaido]

We examined the sera of 587 hunters in Hokkaido (Japan's northernmost island) for the antibody to Borrelia burgdorferi (B. burgdorferi) by enzyme immunoassay, clarified the conditions related to antibody positivity in these subjects according to region, and studied the effects of factors such as age and lifestyle on the antibody titer. In contrast with an anti-B. burgdorferi antibody positive rate of 7.1% in control sera, that in the hunters' sera was 16.0%. Among those positive for the anti-B. burgdorferi antibody, the antibody positive rate in sera excluding those testing positive in the serological test for syphilis was 5.5% in the controls, and 15.4% in the hunters, the latter rate being significantly higher (p less than 0.05). In both hunters and control groups, the antibody-positive rate tended to be higher in older subjects, but the antibody titer showed no correlation with their age, or the duration of their hunting experience. Examination of the hunters' occupations revealed a tendency toward high titers in those engaged in dairy farming. The antibody positivity of those who went gathering edible wild plants was significantly higher than those did not (p less than 0.05). These observations suggested that the high antibody-positive rate in hunters may have been due largely to the effect of activities other than hunting as sources of infection by Borrelia.     

Is there any evidence for an association between ankylosing spondylitis and Borrelia burgdorferi infection?

The suggested relationship between Borrelia burgdorferi and seronegative spondyloarthropathies has been studied in 125 patients with ankylosing spondylitis (AS). IgG antibodies to Borrelia burgdorferi were present in 11 of 125 patients (8.8%) and in 25 of 125 controls (20%). No patient had clinical Lyme borreliosis. HLA-B27 status was known for 82 patients with AS. There was no difference between B27+ and B27 patients. This study provides no evidence that Borrelia burgdorferi is associated with AS.     

Evaluation of the intrathecal antibody response to Borrelia burgdorferi as a diagnostic test for Lyme neuroborreliosis

The intrathecal antibody response to Borrelia burgdorferi was evaluated in American and West German patients with Lyme neuroborreliosis. By an antibody capture enzyme immunoassay, 12 (92%) of 13 patients from the USA with Lyme meningitis were found to have intrathecal antibody production to B. burgdorferi, usually of multiple isotypes, most commonly IgA. Of 12 patients with putative late central nervous system manifestations of Lyme disease, 5 (42%) had local production of IgG or IgA spirochetal antibody, but cerebrospinal fluid (CSF) abnormalities could not be demonstrated in 6 patients with late peripheral nervous system manifestations of the disorder. Compared with American patients, 30 European patients with neuroborreliosis had significantly higher CSF:serum ratios of specific antibody both early and late in the illness. Intrathecal antibody determinations are the most specific diagnostic test currently available for Lyme neuroborreliosis, but local antibody production in CSF is an inconsistent finding in American patients with late neurologic manifestations of the disorder.     

莱姆病螺旋体鞭毛平截性蛋白的表达及其诊断潜力研究

目的原核表达伯氏疏螺旋体鞭毛蛋白Flagellin A基因特异性区段,获得重组鞭毛蛋白平截性蛋白作为诊断抗原,建立间接ELISA方法用于动物莱姆病的诊断。方法 PCR扩增获取伯氏疏螺旋体鞭毛蛋白基因的同源性较低的第394-798bp区段,构建重组质粒pGEX-4T-1/tFlaA,构建好的表达质粒转化到大肠杆菌BL21（DE3）中进行表达,并纯化重组蛋白,用纯化的表达蛋白作为莱姆病诊断的抗原,用于ELISA检测实验感染小鼠莱姆病。结果成功构建莱姆病螺旋体鞭毛平截性蛋白的表达载体,重组蛋白在宿主菌内高效、稳定表达,重组平截性蛋白显示了可作为ELISA诊断的抗原用于莱姆病的诊断价值。结论纯化的伯氏疏螺旋体鞭毛蛋白可作为莱姆病ELISA诊断抗原用于莱姆病的诊断,为莱姆病快速诊断试剂盒的开发打下基础。     

First isolation of Borrelia burgdorferi sensu lato from Ixodes ricinus ticks in Morocco

To determine the infection rate of Ixodes ricinus (I. ricinus) ticks with Borrelia burgdorferi sensu lato (B. burgdorferi sl) and to assess the frequency of the individual Borrelia species in this tick species, a total of 295 I. ricinus were collected in Taza region (Northeast of Morocco), from January to June 2002. The presence of B. burgdorferi sl was determined by direct fluorescence antibody assay (DFA) and by PCR after culture. B. burgdorferi sl isolates were identified at the species level by restriction fragment length polymorphism analysis of amplified products. The mean rate of I. ricinus infection with B. burgdorferi sl was 47.8%. Isolation attempts in BSK II medium resulted in 26 pure isolates. However, PCR performed on culture medium allowed to identify 82 Borrelia DNAs. B. lusitaniae has been identified from 76 out of 82 infected I. ricinus ticks (92.7%). Three ticks were infected by B. burgdorferi ss, and three other ticks were infected by B. garinii. This is the first report of the presence of B. burgdorferi sl in Morocco and more specifically of B. burgdorferi ss in North Africa.     

莱姆病与钩端螺旋体病血清学交叉反应问题的研究

钩端螺旋体病患者与伯氏疏螺旋体偶尔可出现交叉反应,在动物试验亦得到证实,但出现频度和滴度均不高。莱姆病患者血清则未出现钩端螺旋体抗体。     

Gonarthritis, lymphadenopathy and disseminated choroiditis as a primary manifestation of Lyme borreliosis. Case report and discussion of diagnostic possibilities in Lyme borreliosis

A 26-year-old female patient was admitted to the hospital because of bilateral gonarthritis, lymphadenopathy and disseminated chorioiditis as primary manifestation of Lyme borreliosis. Antibody titers against Borrelia burgdorferi did not reach diagnostic levels as determined by an indirect immunofluorescence assay. However, diagnosis was established by the detection of IgG and IgM antibodies in Western blot analysis, and by the demonstration of an enhanced T-cell proliferation to Borrelia burgdorferi in a lymphocyte proliferation assay. This case report indicates that arthritides may already occur in stage II (disseminated infection) of Lyme borreliosis. Therefore, Lyme borreliosis must be considered in patients with chorioiditis and pauciarticular arthritis. In the case of non-diagnostic antibody titers in indirect immunofluorescence tests (or ELISA), Western blot analysis and lymphocyte proliferation assays should be performed in addition.     

The T-cell proliferative assay in the diagnosis of Lyme disease

OBJECTIVE: To determine the sensitivity and specificity of the T-cell proliferative assay as a diagnostic test in Lyme disease. DESIGN: Cross-sectional study of patients with Lyme arthritis or chronic neuroborreliosis who had a history of erythema migrans, positive antibody responses to Borrelia burgdorferi by enzyme-linked immunosorbent assay (ELISA), or both; patients with other diseases; and healthy subjects. SETTING: Diagnostic Lyme disease clinic in a university hospital. PATIENTS: Forty-two of the 67 patients with active Lyme arthritis or chronic neuroborreliosis who were seen during the study period; 16 patients with inactive late Lyme disease; 77 patients with other rheumatologic or neurologic diseases; 9 workers from the Borrelia laboratory; and 9 healthy subjects. MEASUREMENTS AND MAIN RESULTS: Nineteen of 42 patients with Lyme arthritis or chronic neuroborreliosis and 4 of 77 patients with other diseases had positive T-cell proliferative responses to B. burgdorferi antigens. The sensitivity of the proliferative assay was 45% (95% Cl, 30% to 60%) and the specificity was 95% (95% Cl, 87% to 99%). Twelve of 27 patients with active Lyme arthritis, 7 of 15 patients with chronic neuroborreliosis, 4 of 16 patients with inactive Lyme disease, 4 of 9 healthy Borrelia laboratory workers, and 0 of 9 healthy subjects had positive responses. Three of five patients with Lyme disease who had negative or indeterminant antibody responses by ELISA had positive T-cell proliferative responses. CONCLUSION: The T-cell proliferative assay may be a helpful diagnostic test in the small subset of patients with late Lyme disease who have negative or indeterminant antibody responses by ELISA.     

Prevalence of Borrelia burgdorferi sensu lato in ticks, Ixodes ricinus in Tarnowskie Góry district

Borrelia burgdorferi is an aetiological factor of borreliosis (Lyme disease). The main vectors of Borrelia burgdorferi are larvae, nymphs, and females of Ixodes ricinus. The aim of this paper was to analyse infection parameters of Borrelia burgdorferi in a selected populations of Ixodes ricinus. The study was conducted in Tarnowskie Góry administrative district (Krupski M?yn, Zielona, Lubliniec, Tarnowskie Góry, Swierklaniec, Tworóg, and Zbros?awice). A total of 85 ticks were collected with a piece of cloth dragged over the vegetation. The Borrelia burgdorferi infection was confirmed with a PCR method, using flagellin protein gene DNA amplification. DNA of Borrelia burgderferi sensu lato was present in 14 ticks, which constituted 16.5% of the population studied. The percentage of infected females and nymphs was 26.8%, 22.2%, and 5.6%, respectively. A high prevalence of the pathogen (50%) was in ticks revealed in the recreation areas and the community forest in Swierklaniec.     

Antibody response to IR6, a conserved immunodominant region of the VlsE lipoprotein, wanes rapidly after antibiotic treatment of Borrelia burgdorferi infection in experimental animals and in humans

Invariable region (IR)(6), an immunodominant conserved region of VlsE, the antigenic variation protein of Borrelia burgdorferi, is currently used for the serologic diagnosis of Lyme disease in humans and canines. A longitudinal assessment of anti-IR(6) antibody levels in B. burgdorferi-infected rhesus monkeys revealed that this level diminished sharply after antibiotic treatment (within 25 weeks). In contrast, antibody levels to P39 and to whole-cell antigen extracts of B. burgdorferi either remained unchanged or diminished less. A longitudinal analysis in dogs yielded similar results. In humans, the anti-IR(6) antibody titer diminished by a factor of > or =4 in successfully treated patients and by a factor of <4 in treatment-resistant patients. This result suggests that the quantification of anti-IR(6) antibody titer as a function of time should be investigated further as a test to assess response to Lyme disease therapy or to determine whether a B. burgdorferi infection has been eliminated.     

Proliferative responses of mononuclear cells in Lyme disease. Reactivity to Borrelia burgdorferi antigens is greater in joint fluid than in blood

In 27 patients with early Lyme disease, the mean response of peripheral blood mononuclear cells (PBMC) to Lyme spirochetal Borrelia burgdorferi antigens (723 counts per minute) was similar to that of control subjects. During convalescence, 2-3 weeks later, the patients' mean response was significantly higher (2,075 cpm, P less than 0.008). Compared with those with early disease, the PBMC of 22 patients with Lyme arthritis reacted even more to B burgdorferi (2,923 cpm, P less than 0.0004), and, by far, the greatest response was in concomitantly obtained synovial fluid mononuclear cells (15,238 cpm, P less than 0.001). The PBMC of patients with early Lyme disease reacted slightly less to phytohemagglutinin and pokeweed mitogen than those of normal control subjects, but patients with arthritis had greater than normal mitogen responses. In contrast, mitogen reactivity among synovial fluid cells was markedly decreased and correlated inversely with the response to antigen. Thus, in patients with Lyme disease, the antigen-specific responses of mononuclear cells increase as the disease progresses, and in those with arthritis, the greatest reactivity to antigen is found in cells in the inflamed joint.     

Lymphoproliferative responses to Borrelia burgdorferi in Lyme disease

OBJECTIVE: To compare lymphocyte proliferative responses to Borrelia burgdorferi in healthy controls and patients with Lyme disease. PATIENTS: Twelve patients fulfilling case-definition criteria for Lyme disease. Twelve healthy volunteers and two newborns served as controls. MEASUREMENTS: Antibodies to B. burgdorferi were measured by enzyme-linked immunosorbent assay (ELISA). Proliferation of peripheral blood lymphocytes cultured for 5 days with B. burgdorferi, recall antigens, or pokeweed mitogen was measured by radioactive thymidine uptake. RESULTS: Lymphocytes from 11 patients with Lyme disease, 8 healthy seronegative controls, and two newborns showed elevated responses when stimulated with B. burgdorferi. When a patient and a control were studied on the same day, the patient's lymphocyte response to B. burgdorferi exceeded the control's in only 5 of 12 cases. Lymphocytes from both patients and controls responded to B. burgdorferi isolates from three different sources. CONCLUSIONS: Heightened lymphocyte responses to B. burgdorferi are found in patients with Lyme disease but elevated responses also frequently occur in healthy controls. At present, the interpretation of a positive lymphocyte response to B. burgdorferi would be difficult in ambiguous clinical situations.     

从全沟硬蜱分离的伯氏疏螺旋体的实验研究

本文报道了从全沟硬蜱分离的一株伯氏疏螺旋体(VL株)实验研究的结果。该株螺旋体同莱姆病螺旋体标准株在超微结构上相近,可以和高稀释度的抗伯氏疏螺旋体抗体发生间接免疫荧光反应。SDS-PAGE电泳结果显示其蛋白组成和标准株的蛋白图谱相同。热变性温度法测定其DNA的G+Cmol％含量为28.1％,和标准株的含量无明显区别,试验结果证实此株螺旋体属伯氏疏螺旋体。