Virulent Toxoplasma gondii strain RH promotes T-cell-independent overproduction of proinflammatory cytokines IL12 and gamma-interferon.

The aim of this study was the analysis of the cytokine response in BALB/c mice infected with the highly virulent RH or the weakly virulent Beverley strains of Toxoplasma gondii. Analysis of cytokine messages showed increased expression of IL12, IFN-gamma and TNF-alpha, but not IL4 mRNAs in spleen cells after infection with the T. gondii strains RH and Beverley. High levels of circulating IL12 and IFN-gamma were detected in the serum of mice infected with strain RH, although TNF-alpha levels remained low. In contrast, the same cytokines were detected at only low levels in the serum of mice infected with the Beverley strain. Administration of antibody against IL12 or IFN-gamma significantly delayed time to death of mice infected with strain RH compared to controls. T-Cell-deficient as well as normal mice were equally infected by strain RH, suggesting that T lymphocytes do not contribute to the response. Depletion of natural killer cells from the splenocyte population abolished the in vitro production of IFN-gamma. Together, our data suggest that the virulent strain RH induces in BALB/c mice a type 1 cytokine pattern with T-cell-independent overproduction of IL12 and IFN-gamma that may be involved in the pathogenesis of this micro-organism.

Human Granulocytic Ehrlichiosis in Belgium: an underestimated cause of disease.

OBJECTIVES: Human Granulocytic Ehrlichiosis (HGE) is a recently discovered zoonosis and, in Europe, not always included in laboratory testing when a patient presents with a history of tick bite. The available serology results indicate that HGE should be included in the screening panel when a tick-borne disease is suspected. METHODS: Serological methods were applied; i.e. indirect immunofluorescence and Western Blot analysis. Sixty-five serum samples from 47 patients were analysed, of six patients sequential samples were available. RESULTS: 33.8% of the submitted samples were found positive in indirect immunofluorescence, Western Blot confirmed 46.1% of these positive samples. CONCLUSIONS: Although the causative agent and the vector for HGE, Ixodes ticks, are present in Belgium, serology for HGE is seldom solicited. Ehrlichiosis is apparently not always considered as a plausible or possible cause for illness, even when the patient presents with a history of tick bite. We present here a, true be it, incomplete picture of the present situation in Belgium, but nevertheless indicating that it is warranted to test patients with a history of tick bite not only for Lyme disease, but also for HGE.

Seroprevalence of human granulocytic ehrlichiosis infection in Belgium.

In order to determine the prevalence of human granulocytic ehrlichiosis (HGE) in Belgium, the sera of 216 patients previously diagnosed with Borrelia burgdorferi infection were analysed for possible coinfection with the agent of HGE. For this purpose, an indirect immunofluorescence assay was applied, and positive results were confirmed by Western blot using a 44-kilodalton recombinant protein (rP44) specific for the agent of HGE. Sixteen of the 216 (7.4%) sera tested were positive for the HGE agent using indirect immunofluorescence assay, and seven (3%) of them were confirmed positive by Western blot. These data suggest the agent for HGE is present in Belgium and may cause coinfection in patients infected with Borrelia burgdorferi, as has been reported in the USA and elsewhere in Europe. This is the first report documenting the identification of this agent in Belgium.

[Followup of pregnant women in the context of malaria: two case reports]. / Suivi de la grossesse dans le cadre du Paludisme: à propos de deux observations.

Whether or not a pregnant women should travel to regions where malaria is highly endemic will always be open to question as no prophlaxis can guarantee complete protection in every case. No chemoprophylaxis is 100% effective or entirely without side-effects, particularly for pregnant women whose immune status provides a favourable environment for the emergence of this parasitosis. The advice given will depend on the country and the region, a possible resistance of the parasite to treatment, the season, the length and circumstances of the stay and individual factors. The choice of treatment is very limited as many therapies are contra-indicated in pregnancy; in addition it is extremely important to give a sufficiently clear explanation concerning precautions and prophylaxis as well as of how a possible bout of malaria should be treated, particularly in case of extended stay.

Distinct requirements for IL-6 in polyclonal and specific Ig production induced by microorganisms.

The role of IL-6 in Ig production induced in the mouse by lactate dehydrogenase-elevating virus (LDV), Toxoplasma gondii or lipopolysaccharide (LPS) was assessed. Following infection with LDV, a strong activator of B cells, an early and transient IL-6 production was observed, that originated predominantly from macrophages. Whereas LDV-induced B lymphocyte proliferation appeared independent of IL-6, mice deficient for this cytokine showed a marked reduction in their total T-dependent IgG2a production when compared to their normal counterparts. By contrast, specific responses directed against either LDV or non-viral antigens administered at the time of infection were not decreased in the absence of IL-6. Similarly, polyclonal, but not anti-parasite IgG2a production triggered by T. gondii infection was strongly dependent on the presence of IL-6. Finally, T-independent total IgG3 secretion triggered by LPS was also markedly reduced in IL-6-deficient mice. These results suggest that IL-6 plays a major role in T-dependent and T-independent polyclonal Ig production following B lymphocyte activation by viruses, and parasites, but not in specific antibody responses induced by the same microorganisms.

IFN-gamma-independent IgG2a production in mice infected with viruses and parasites.

After infection with some viruses and intracellular parasites, antibody production is restricted to IgG2a. We first observed that, whereas live viruses such as lactate dehydrogenase-elevating virus (LDV) or mouse adenovirus induced mostly an IgG2a response, a large proportion of antibodies produced against killed viruses were IgG1. This IgG1 antiviral response was suppressed when live virions were added to inactivated viral particles. These results indicate that the IgG2a preponderance is related to the infectious process itself rather than to the type of antigen involved. Since IFN-gamma is known to stimulate IgG2a production by activated B lymphocytes and to be secreted after infection, we examined the role of this cytokine in the antibody isotypic distribution caused by LDV. Most IgG2a responses were relatively unaffected in mice deficient for the IFN-gamma receptor or treated with anti-IFN-gamma antibody. A similar IFN-gamma-independent IgG2a secretion was observed after infection with the parasites Toxoplasma gondii and Trypanosoma cruzi. However, the IFN-gamma-independent IgG2a production triggered by infection still required the presence of functional T(h) lymphocytes. Therefore, signal(s) other than IFN-gamma secretion may explain the T(h)-dependent isotypic bias in antibody secretion triggered by viruses and parasites.

Eosinophilic pleocytosis and myelitis related to Toxocara canis infection.

Toxocara canis causes the visceral larva migrans syndrome in which central nervous involvement is rare. We report the case of a 40-year-old woman presenting with a subacute weakness of the right leg and dysaesthesiae in the right Th8-Th10 dermatomas. Spinal magnetic resonance imaging examination showed abnormal hyperintensity within the spinal cord. Cerebrospinal fluid analysis revealed eosinophilic pleocytosis. Antibody titres to Toxocara canis were higher in the cerebrospinal fluid than in the serum. Treatment using mebendazole led to a complete clinical recovery, normalization of cerebrospinal fluid parameters and improvement in spinal magnetic resonance imaging abnormalities.

Cervical lymphadenitis and cat scratch disease (CSD): an overlooked disease?

Cat Scratch Disease (CSD) is a benign disease characterized by regional lymphadenopathy affecting most frequently the head and neck region in children and young adults. In the present paper, the authors describe four cases of CSD focusing on clinical history, diagnostic management and therapy. The main germ responsible for this lymph node disease is Bartonella henselae. Diagnosis is based on history, serology and histological findings. Clinical evolution is generally favorable despite the fact that complications occur in about 5% to 13% of patients including encephalitis, hepatitis and Parinaud's oculoglandular syndrome. Antibiotic treatment is only considered for highly symptomatic patients. Surgical excision of the lymphadenopathy is useful to establish the diagnosis when serology is not available and/or when the adenopathy become fluctuating. The authors emphasize the increasing incidence of patients with CSD in the ENT population and the algorithm for CSD disease affecting the cervical lymph nodes.

Acute and chronic phases of Toxoplasma gondii infection in mice modulate the host immune responses.

Murine antibody responses to soluble proteins are generally restricted to the immunoglobulin G1 (IgG1) isotype. When mice were infected with Toxoplasma gondii Beverley and concomitantly immunized with a soluble unrelated protein antigen, a modification in the isotypic distribution of antibodies directed against this nonparasite antigen was observed, with a preferential production of IgG2a. Interestingly, when mice were immunized with a soluble protein antigen during the chronic phase (day 40) of infection with T. gondii Beverley, a similar modification in the isotypic distribution of antiprotein antibodies was observed.

Detection of Toxoplasma gondii tachyzoites and bradyzoites in blood, urine, and brains of infected mice.

Different techniques for identifying Toxoplasma gondii were compared. PCR was used to amplify part of the major surface antigen P30 gene of T. gondii. Amplified-DNA detection with the DNA enzyme immunoassay (PCR-DEIA) was more sensitive than ethidium bromide staining after agarose gel electrophoresis and as sensitive as nested PCR. PCR-DEIA, using common enzyme-linked immunosorbent assay (ELISA) methods, avoids agarose gel electrophoresis for the identification of amplified products. T. gondii can also be detected with equal sensitivity in infected fibroblasts, but only after at least 8 days of cell culture. PCR-DEIA is thus recommended because of its sensitivity and convenience for detecting early parasitemia in the surveillance of toxoplasmosis among pregnant women and immunocompromised hosts. The courses of infection in mice infected with two strains of T. gondii were compared. Tachyzoites of the virulent strain T. gondii RH, killing the host in 4 days, were identified in urine specimens and blood samples of mice 24 to 94 h after inoculation but not in brains, but no antibodies were detected. After intraperitoneal inoculation with cysts of the low-level virulence Beverley strain of T. gondii, parasites were identified in blood samples 4 days later and up to 17 days (but not in urine specimens) and in the brain from day 6 through day 525. By ELISA, high antibody titers were found from day 11 to day 525, with parasitemia preceding the appearance of antibodies. The usefulness of PCR-DEIA tests in conjunction with the search for circulating antibodies for the early diagnosis of toxoplasmosis in humans is discussed.

MRI of cervical cord lesions and their resolution in Toxocara canis myelopathy.

We report serial MRI findings in a 58-year-old man with cervical cord involvement by Toxocara canis, in whom antihelminthic chemotherapy yielded improvement of the neurological deficits and cord lesions seen on MRI.

Serodiagnosis of Lyme borreliosis by Borrelia burgdorferi sensu stricto, B. garinii, and B. afzelii western blots (immunoblots).

The performance of Western blots (immunoblots) prepared with eight strains of Borrelia burgdorferi representing B. burgdorferi sensu stricto, B. garinii, and B. afzelii genospecies was tested with a panel of sera with various clinical presentations collected from eight geographic regions. European sera were generally more reactive to blots prepared with B. garinii or B. afzelii strain antigens, in particular B. garinii 20047 and B. afzelii VS461. North American sera were more reactive with B. burgdorferi sensu stricto strains. Our observation of significant differences in the levels of reactivity of some sera on Western blots of certain strains is potentially important for the development and implementation of generic interpretive criteria. Preferential reactivity of sera from patients with nerve and/or palsy symptoms to B. garinii strains and with cutaneous disease to B. afzelii strains was observed. On the basis of our results, we have concluded that strain 20047 is the best strain to use for the development of a generic Lyme borreliosis Western blot for Europe.

Simultaneous presence of different Borrelia burgdorferi genospecies in biological fluids of Lyme disease patients.

Oligonucleotide primers based on Borrelia burgdorferi sensu lato ospA gene sequences have been designed for use in the PCR to type all (SL primers) or each (GI to GIII primers) of the B. burgdorferi sensu lato genospecies involved in Lyme disease. These genospecies-specific primers were then used in the PCR on 24 biological fluids collected from 18 neuroborreliosis patients. Among the samples tested, 20 contained DNA from Borrelia garinii, 11 contained DNA from B. burgdorferi sensu stricto, and 10 contained DNA from Borrelia afzelii. In toto, 10 patients appeared to have been infected by a single genospecies and 8 were infected by more than one Lyme disease-associated genospecies. Serum specimens from six patients were absorbed with heterologous antigens and tested by Western blotting (immunoblotting). In four cases, residual immunodetection revealed specific epitopes of genospecies also detected by PCR; in two of them, the concordant results indicated pluri-infection of the patients. In the other two cases, Western blotting showed specific antibodies for two genospecies of Borrelia, while PCR detected DNA from only one. In summary, the data underscored the relatively high prevalence of pluri-infections in Lyme disease and confirmed the association of B. garinii with neuroborreliosis.

Canine Lyme disease in Belgium.

Lyme borreliosis is a multisystem disease caused by the tick-borne spirochaete Borrelia burgdorferi. In addition to causing human illness, borreliosis has been recorded in many species of mammals, including domestic animals which come into contact with the infected tick vector. This paper describes the first two cases to the authors' knowledge of Lyme borreliosis in dogs in Belgium. Both animals suffered recurrent lameness and general prostration. In addition, one of the dogs had recurrent paralysis of nerves V, VII, IX and X, leading to dysphagia and total bilateral paralysis of the mandibular muscles. This complication of Lyme borreliosis has been well documented in human pathology, but has not previously been recorded in the veterinary literature.

[Risk of contamination by Borrelia burgdorferi S. lato in a forest environment. Survey during 13 months of abundance of the tick Ixodes ricinus and its level of infestation by the Lyme borreliosis agent in Brittany]. / Risque de contamination par Borrelia burgdorferi S. lato en milieu forestier. Suivi pendant 13 mois de l'abondance de la tique Ixodes ricinus et de son niveau d'infestation par l'agent de la borréliose de Lyme en Bretagne.

The authors, in a forest in Brittany previously studied for several years, caught by flagging, each month from April 1992 to May 1993, nymphs of I. ricinus tick, and looked by indirect immunofluorescence, for B. burgdorferi infestation. An amount of 1,506 ticks was thus studied. Infestation frequency was varying from 0 per cent in January and February to 14.4 per cent in August. Standarding of tick collecting method allowed to establish, for each month, a tick, "availability" index, and, according to the spirochete infestation frequency, to do estimation of the risk level, for human visiting the concerned forest, of being infected by B. burgdorferi. Obtained results show that this risk is the highest in August, and quite non-existent in January and February.

Evidence for the involvement of different genospecies of Borrelia in the clinical outcome of Lyme disease in Belgium.

In addition ot Borrelia burgdorferi, recognized as the aetiological agent of Lyme disease, at least two separate genospecies have recently been described. A relationship between infection by strains belonging to different genospecies and clinical outcome has been suspected. In this paper, 9 cases of Lyme arthritis were attributed to infection by B. burgdorferi sensu stricto, 18 cases of neuroborreliosis to B. garinii and one case of acrodermatitis chronica atrophicans to a strain of B. afzelii. These conclusions were based on the preferential reactivity of sera with antigens of given strains in Western blots and on residual reactivity after absorption of sera with antigens of representative strains. No conclusion could be reached concerning sera of 10 patients with erythema migrans.

Polyclonal and oligoclonal IgA synthesis in the cerebrospinal fluid of neurological patients: an immunoaffinity-mediated capillary blot study.

An intrathecal synthesis of IgA has been reported in various neurological disorders. However, the frequency of its occurrence and the electrophoretic characteristics of the locally produced IgA remained a matter of controversy. We developed a sensitive immunoaffinity-mediated capillary blot technique for the detection of polyclonal and oligoclonal IgA in the CSF of 115 patients with various neurological disorders. Paired CSF and serum samples containing 50 ng IgA after appropriate dilutions were submitted to isoelectric focusing in agarose gels; IgA was then blotted onto a polyvinylidene difluoride sheet coated by an anti-IgA antiserum or by infectious antigens. The immunoblots were revealed by an alkaline phosphatase-conjugated anti-IgA antiserum. Only five samples displayed CSF-restricted oligoclonal IgA bands, including two out of 33 from MS patients. In herpetic encephalitis (n = 5) and varicella-zoster meningitis (n = 2), a strong intrathecal production of virus-specific IgA antibodies was detectable. In such cases, faint oligoclonal IgA antibodies were superimposed on a polyclonal background. A weak local production of anti-Borrelia burgdorferi IgA antibodies was present in two out of four cases of neuroborreliosis.

[Epilepsy disclosing neuroborreliosis]. / Epilepsie révélatrice d'une neuroborréliose.

After two seizures, a 13 year-old boy experienced headache, fatigue and loss of appetite over a period of 3 weeks. There was a bilateral papilledema with normal visual acuity. CT and MRI disclosed two ischemic foci, that were interpreted as evidence of vasculitis. High serum levels of IgG and IgM antibodies specific to Borrelia burgdorferi, were present. The patient had attended an outdoor scout camp in a area, in south-east Belgium, known to be endemic for tick-born borreliosis. The clinical symptoms, the levels of the specific antibodies and the radiologic abnormalities responded dramatically to treatment. We believe that seizures in this case were related to cerebral vasculitis. This case confirms the extreme diversity of the neurological manifestations of Borreliosis.