Electron microscopy and the polymerase chain reaction of spirochetes from the blood of patients with Lyme disease.

Results of studies using direct antigen detection suggest that seronegative Lyme borreliosis is not rare and support the hypothesis that Borrelia antigens can persist in humans. We report three successful cultures from blood out of 30 attempts from 96 Lyme disease patients. The proof of borreliaemia in early or late phases of Lyme disease by immuno-capture electron microscopy has practical importance for subsequent cultivation. The polymerase chain reaction with oligonucleotide sequences directed against 16S rRNA identified two of our blood isolates as Borrelia burgdorferi genospecies III., VS 461 group, and one as Borrelia garinii sp. nov. All of the three isolates were reactive with monoclonal antibody H9724 against flagellin and with antibody against main extracellular protein at 83 kDa. Borrelia garinii had a single predominant protein OspA at 33.5 kDa and reacted with monoclonal antibody H5332 in contrast to two isolates of the VS 461 group with two major proteins OspA and OspB at 32.5 and 35 kDa. We conclude that isolation of spirochetes from the blood might prove successful in clinically selected cases of Lyme borreliosis. Immuno-capture electron microscopy has proved to be a sensitive assay for monitoring and studying Lyme borreliosis.

[The clinical picture and etiology of meningoencephalitis in hospitalized patients in 1986]. / Klinický obraz a etiologie meningoencefalitid u hospitalizovaných pacientu v roce 1986.

In 1986 in the CSR 978 cases of acute meningoencephalitis were notified, incl. 184 in the capital. In a group of 129 patients aged 1 to 63 years hospitalized at the First Clinic for Infectious Diseases in Prague-Bulovka (Paediatric Faculty Charles University) in 1986 the authors investigated the clinical course of acute meningoencephalitis in relation to the causal agent of the disease. The aetiological agent was assessed by isolation from cerebrospinal fluid or by isolation from faeces with the corresponding antibody response, or by serological examination only. In children under 15 years of age the etiological agent was assessed in 36%, in adults in 65%. In the investigated group, contrary to previous years (3), tick-borne encephalitis was involved which accounted for the majority of severe cases of the disease. The severe clinical course, manifested at first by symptoms of cerebral oedema with impaired consciousness, is more frequent with advancing age. In adults tick-borne encephalitis had a severe course in one third of the patients. In the other age groups the development was favourable, in particular in the group of children aged 4-10 years. As compared with previous years, there was no fatal case, no permanent paresis as a complication of the disease (2).

[Immunologic study of Lyme borreliosis]. / Imunologická studie lymeské borreliózy.

In 84 patients in the second or third stage of serologically proved Lyme borreliosis suffering from different forms of central and/or peripheral nervous system involvement the participation of autoimmunological mechanisms was investigated. The cellular hypersensitivity to encephalitogenic basic protein and to antigens from peripheral myelin was evaluated on the cytopherometer Opton according to the influence of macrophage slowing factor--liberated during the short-term incubation of the sensibilized lymphocytes with specific antigen--on the mobility of tanned sheep red blood cells in the electrical field. The frequency of positive findings in comparison with the results in other types of neuroinfections, examined previously, put the Lyme borreliosis on the first place. The explanation of this fact may be seen in the immunoregulatory disturbances, even though the changes of the total and active T lymphocytes were not significant. The knowledge of immunological changes is very important for the indication of immunomodulating procedures suitably complementing the treatment by antibiotics.

[Arrhythmia as a major manifestation of Lyme borreliosis]. / Arytmie jako vedoucí projev lymeské borreliózy.

The submitted case-history is focused on one of the leading manifestations of cardiac affection in Lyme borreliosis. The authors recapitulate and analyze the patient's complaints, her electrocardiographic and other findings and confront them with data from the literature on this "new" nosological unit.

Cross reactivity among human enterovirus serotypes as revealed by microneutralization assay technique.

Paired or tripled serum specimens examined for cross reactions between poliomyelitis viruses and other non-polio enteroviruses were obtained from 356 patients (217 males and 139 females) with various clinical diagnoses (aseptic meningitis, meningoencephalitis, paretic neuro-infections, upper respiratory tract infections, bronchopneumonia) who were sampled in 1984 and 1985. The sera were examined by the microneutralization assay technique in flat-bottomed 96-well polyvinyl chloride plates against 100-300 TCD50 of these enterovirus serotypes: CA9, CB1-CB5, E1, E2, E4-E9, E17, E20, E24, E30, EV71 and polioviruses P1-P3. The tests were carried out on BGM or RD cells. Confirmed significant seroconversions to at least one enterovirus serotype were observed in 87 (24.2%) patients. Of these, 27 (31%) reacted by parallel significant or insignificant increases in the heterotypic antibody titre to poliovirus, which accounted for 7.6% of all patients examined. Heterotypic antibody responses may become source of errors in individuals examined for poliovaccine efficacy or the state of specific immunity response to polio, but are unlikely to discredit the overall outcomes of the seroepidemiological surveys conducted in the general population.

An immunological study of Lyme disease.

84 patients in the second or third stages of serologically diagnosed Lyme disease suffering from different forms of central as well as peripheral nervous system involvement were tested for the participation of autoimmune mechanisms. Cell hypersensitivity to the encephalitogenic basic protein and to antigens from peripheral myelin was evaluated on an Opton cytopherometer according to the effect of the macrophage slowing factor--liberated during short-term incubation of sensibilized lymphocytes by the respective antigen--on the mobility of tannin-treated sheep red blood cells in an electric field. Judged by the presence of autoimmune reactions. Lyme disease has come to occupy a prominent position compared with previously examined other types of neuroinfections. An explanation can be sought in disordered immunoregulating mechanisms, even though the changes in total and active T lymphocytes were not statistically significant. Data on immunological changes are relevant for the indication of immunomodulating therapy as a suitable complement to antibiotic treatment.

Ceftriaxone in the treatment of Lyme neuroborreliosis.

In an open non-comparative clinical trial with the aim of evaluating the clinical efficacy and safety of a 14 day course of 2 g ceftriaxone once daily, 46 patients with neuroborreliosis were entered at the Infectious Diseases Teaching Hospital in Prague 8. In 39 patients the diagnosis was early Lyme neuroborreliosis. Seven patients suffered from late stage disease. Clinical results were 30% of patients cured at the end of treatment and 85% after 9 months in early stage disease. In late stage disease two patients out of seven were cured and four had improved after 12 months. One patient died because of cardiac infarction. In no patient had treatment to be discontinued because of adverse reactions to antibiotics.

Electron microscopy of Langerhans cells and Borrelia burgdorferi in Lyme disease patients.

To investigate dermal and epidermal involvement in the presence of Borrelia burgdorferi and to analyze the role of Langerhans cells and keratinocytes, 14 cases of erythema chronicum migrans and two controls were studied by means of electron microscopy, using negative staining and sectioning techniques. Using immunoelectron microscopy and histochemistry, positive results for B. burgdorferi were disclosed in 5 cases of erythema chronicum migrans and 3 cases of neuroborreliosis which were confirmed by cultivation. We cultured 4 stains of B. burgdorferi from the skin, 1 from blood and 2 from cerebrospinal fluid in BSK medium. Near to the centre of erythema chronicum migrans with focal necrosis were both a dissolved basal membrane and keratinocyte desmosomes surrounding damaged B. burgdorferi cells in the epidermis. Markedly oedematous keratinocytes and Langerhans cells with B. burgdorferi were released into lymphocyte infiltrates. At the periphery of all erythema chronicum migrans lesions, keratinocytes were well preserved while all dendritic cells seemed to be vacuolated. Above foci of B. burgdorferi located perivascular or among collagen fibers, Langerhans cells were frequent and more granulated. The possible role of Langerhans cells in the identification and elimination of B. burgdorferi is discussed.