Characterization of the cellular and humoral immune response to outer surface protein C and outer surface protein 17 in children with early disseminated Lyme borreliosis.

OspC and Osp17 are immunodominant proteins of Borrelia burgdorferi eliciting a clear humoral immune response in adult patients with systemic Lyme disease. In this study, the cellular immune response to B. burgdorferi and the major outer surface proteins OspC and Osp17 was investigated in children during the course of early disseminated B. burgdorferi infection. Lymphoproliferative responses to recombinant proteins were compared to the protein-specific humoral immune reaction. OspC induced a clear antibody response but elicited an even stronger cellular immune response. In contrast, a cellular as well as humoral immune reaction to Osp17 was only rarely detected. Follow-up examinations demonstrated that the lymphoproliferative response to B. burgdorferi and OspC persisted for several months after antibiotic therapy. Here, we show that in early disseminated Lyme disease of childhood, OspC is a potent antigen influencing both the humoral and cellular immunity, while Osp17 plays only a minor role in immune activation.

Human herpes virus type 7 DNA in the cerebrospinal fluid of children with central nervous system diseases.

UNLABELLED: Human herpes virus type 7 (HHV-7) has been associated with unspecific febrile syndrome, exanthem subitum (ES), viral rashes and Epstein-Barr virus (EBV) like syndrome. Neurological complications such as hemiplegia or seizures have been described in a few children with ES. Whether HHV-7 may also affect the CNS in the absence of ES is unknown. In this study, we investigated CSF samples from children with different neurological diseases for the presence of HHV-7 specific DNA. A HHV-7 specific nested polymerase chain reaction (PCR) was established amplifying a 478 bp DNA sequence of the glycoprotein U23 of HHV-7 strain SB. 68 children with CNS diseases with inflammatory CSF findings (n = 24), CNS diseases without inflammatory CSF findings (n = 18) and febrile seizures (n = 26) were examined. A total of 26 children with infectious diseases in the absence of neurological disease and 11 children without signs of a peripheral infection and without neurological disease served as controls. The CSF samples of six children from the study groups were HHV-7 PCR positive, but none from the controls. These children were diagnosed with aseptic meningitis (n = 1), viral encephalitis/meningoencephalitis (n = 2), facial palsy (n = 1), vestibular neuritis (n = 1) and febrile seizure (n = 1). CONCLUSION: These results indicate that human herpes virus type 7 infection is associated with central nervous system disease in children and should be considered in children whether inflammation in the cerebrospinal fluid is present or not.

IL-6 in CSF during ventriculitis in preterm infants with posthemorrhagic hydrocephalus.

Treatment of infants with posthemorrhagic hydrocephalus by diversion of CSF is frequently complicated by bacterial ventriculitis. We report the CSF values before and during bacterial ventriculitis of four very low birth weight infants with progressive posthemorrhagic ventricular dilatation. Extremely high CSF IL-6 concentrations of between 8,000 and 61,000 pg/ml were observed and compared with values reported in the literature. IL-6 seems to be a useful marker for bacterial ventriculitis in preterm infants. The role of IL-6 monitoring in the CSF of preterm infants with posthemorrhagic hydrocephalus for early diagnosis of bacterial ventriculitis prior to clinical manifestation should be clarified by further studies.

Cross-reactivity of Borrelia burgdorferi and myelin basic protein-specific T cells is not observed in borrelial encephalomyelitis.

Borrelial encephalomyelitis, a rare manifestation of Lyme borreliosis, may present as a multiple sclerosis (MS)-like disease. It is postulated that in MS, inflammation of the white matter is caused by a T-cell response directed to myelin antigens. Here, we examined whether a T-cell autoimmune response may play a pathogenetic role in Borrelia-associated white matter disease mediated by cross-reactivity between myelin basic protein (MBP) and B. burgdorferi. We generated a total of 1760 short-term T-cell lines against B. burgdorferi or MBP from two patients with Borrelial encephalomyelitis and compared these with three patients with late Lyme disease, one patient with transverse myelitis, eight patients with MS, and four healthy controls. While a few T-cell lines recognized both B. burgdorferi and MBP, T-cell clones from these lines responded only to the antigen of the original stimulation. Thus, our data do not provide evidence for cross-reactivity between MBP and B. burgdorferi.

Myelin basic protein reactive Th2 T cells are found in acute disseminated encephalomyelitis.

Acute disseminated encephalomyelitis (ADEM), a postinfectious illness of the central nervous system (CNS), is thought to be an autoimmune disease. Here, we characterized the cytokines secreted by myelin-reactive T cells generated from patients with ADEM. The frequency of MBP-reactive T cell lines was ten-fold higher in patients with ADEM compared to patients with encephalitis and normal subjects. Whereas there was no significant IFN-gamma secretion, the predominant cytokine secreted by MBP-reactive T cell lines was IL-4 in patients with ADEM. In contrast, IL-4 secretion was only rarely detected in the controls. The presence of high frequencies of MBP-reactive IL-4 secreting T cells in subjects with ADEM during their recovery phase may be similar to myelin reactive IL-4 secreting T cells observed during the spontaneous recovery of animals with EAE.

The cellular immune response to Bordetella pertussis in two children with whooping cough.

There is increasing evidence that the cellular immune response to Bordetella pertussis plays an important role in the immune protection. Particularly in animal models, Bordetella pertussis-specific T-cells have been shown to confer immunity. In this case report, we therefore investigated the cellular immune response to whole cell Bordetella pertussis bacteria, to the pertussis antigens filamentous hemagglutinin and pertussis toxoid defined by lymphoproliferation and cytokine secretion. Two children with whooping cough were compared to three individuals vaccinated against whooping cough with a whole cell pertussis vaccine. In contrast to the vaccinated controls, the cellular immune response to Bordetella pertussis in children with whooping cough was characterized by a strong proliferation of T cells to whole pertussis bacteria as well as to filamentous hemagglutinin and pertussis toxoid. This response was defined by a marked Th-1 type T cell response with IFN-gamma secretion to all Bordetella pertussis antigens. However, in the control individuals IFN-gamma was secreted only to whole cell Bordetella pertussis bacteria and filamentous hemagglutinin but not to pertussis toxoid. A Th-2 type cytokine response could not be detected in any condition. Our observations suggest that in the immune defense of a natural Bordetella pertussis infection, the Th-1 specific T cell response to filamentous hemagglutinin and particularly to pertussis toxoid may play a major role.

Identification of a T cell subset capable of both IFN-gamma and IL-10 secretion in patients with chronic Borrelia burgdorferi infection.

A novel population of both IFN-gamma- and IL-10-secreting human T cells that differentiate in the presence of exogenous IL-12 in vitro has recently been described. Whether this T cell population exists in vivo is unknown. Borrelia burgdorferi, the etiologic agent of Lyme disease, can induce a chronic infection in the presence of a vigorous humoral immune response. We established T cell lines specific for B. burgdorferi and tetanus toxoid from subjects with chronic B. burgdorferi infection and healthy controls in limiting dilution experiments and assessed proliferation and cytokine secretion. As expected, higher frequencies of B. burgdorferi-specific precursor T cells were observed in Lyme patients compared with controls. In both groups of subjects, T cell lines specific for B. burgdorferi secreted high amounts of IFN-gamma. However, in patients with Lyme disease, 27% of T cell lines secreted not only IFN-gamma but also IL-10, which was only observed in 0.6% of B. burgdorferi-reactive T cell lines generated from controls and in none of the tetanus toxoid-reactive T cell lines generated from either Lyme patients and controls. Single cell PHA cloning confirmed that both cytokines were secreted from one clonally expanded precursor cell. Whole mononuclear cells from B. burgdorferi-infected individuals, but not from controls, secreted IL-12. Moreover, neutralizing anti-IL-12 mAbs inhibited the generation of the IFN-gamma/IL-10 population. These data demonstrate that this novel population of IL-12-induced IFN-gamma/IL-10-secreting T cells is generated in response to chronic B. burgdorferi infection.

Adenovirus infection in cystic fibrosis patients: implications for the use of adenoviral vectors for gene transfer.

Clinical trials using replication-deficient adenovirus as vectors for gene transfer into the airways of cystic fibrosis (CF) patients are in progress. However, little is known about the prevalence of wild-type adenovirus infections in patients with cystic fibrosis and their effect on lung function. To answer these questions, serum IgG and IgM antibody titers against adenovirus type 5 were prospectively measured by an indirect immunofluorescence assay in 199 CF outpatients and in a control group of 45 healthy children and young adults. In addition, we performed pulmonary function tests when the patients were in stable clinical condition. IgM antibodies against adenovirus were present in 104 of the 199 cystic fibrosis patients (52.3%). IgG antibodies against adenovirus were detected in 192 of the 199 cystic fibrosis patients (96.5%), and were significantly higher in cystic fibrosis patients older than 7 years than in younger patients and in age matched controls. IgG antibody titers measured a second time 11.8 months later in 143 of the 199 patients had increased in 48 (33.6%) patients. In 27 of these 48 patients, who had at least a 2-fold increase in antibody titer, FVC and FEV1 decreased by 9.8% (p < 0.05) and 8.3% (p = 0.05), respectively, over 45 months. In a comparison group matched for age, sex, and chronic Pseudomonas aeruginosa infection but no increase in antibody titers, FVC and FEV1 were unchanged. The results indicate that wild-type adenovirus infections are prevalent in cystic fibrosis patients and that wild-type adenovirus infections in cystic fibrosis patients seem to be associated with deterioration in lung function. These observations may have important implications for efficacy and safety considerations when using adenoviral vectors for gene therapy.

Detection of human coronavirus 229E-specific antibodies using recombinant fusion proteins.

Human coronaviruses are known to be a common cause of respiratory infections in man. However, the diagnosis of human coronavirus infections is not carried out routinely, primarily because the isolation and propagation of these viruses in tissue culture is difficult and time consuming. The aim of this study was to evaluate the use of recombinant, bacterial expressed proteins in the serodiagnosis of coronavirus infections. Two proteins were examined: the human coronavirus 229E nucleocapsid protein (N), expressed as a fusion protein in the vector pUR and the coronavirus 229E surface glycoprotein (S), expressed as a fusion protein in the vector pROS. The recombinant proteins were used as antigens in Western blot (WB) assays to detect the 229E-specific IgG antibodies and the results were compared with a standard serological method, indirect immunofluorescence. Serum samples of 51 paediatric patients, suffering from acute respiratory illness, and 10 adults, voluntarily infected with human coronavirus, were tested. The serum samples of the adult group had coronavirus-specific IgG antibodies in both test systems. In contrast, only 8/51 sera of the paediatric group were positive for coronavirus-specific IgG by both WB and IF and 20/51 sera were positive by WB, but not by IF. The overall incidence of human coronavirus infections in the paediatric age group was 55% evaluated by WB analysis and 16% evaluated by IF. This study shows that recombinant human coronavirus 229E proteins are suitable reagents for the epidemiological screening of coronavirus 229E infections.

Antibody reactivity to individual structural proteins of measles virus in the CSF of SSPE and MS patients.

BACKGROUND: Chronic progressive disorders of the central nervous system (CNS) impose diagnostic problems, particularly in younger patients. The demonstration of antibodies against measles virus (MV) in the cerebrospinal fluid (CSF) plays a major role in the laboratory diagnosis of subacute sclerosing panencephalitis (SSPE) as well as multiple sclerosis (MS). OBJECTIVES: Because intrathecally synthesized antibodies against MV can be found in both diseases, it is necessary to establish easy and reliable methods to improve the differential diagnosis. STUDY DESIGN: Seventy-one paired serum/CSF samples obtained from patients with the diagnosis of SSPE (n = 23), MS (n = 14), or acute postinfectious measles encephalitis (APME, n = 8) have been examined. The reactivity of intrathecally synthesized immunoglobulin to individual recombinant MV structural proteins was assessed using Western blot analysis, ELISA as well as isoelectric focusing (IEF). RESULTS: All CSF samples obtained from patients suffering from SSPE showed a strong antibody response to MV-nucleocapsid (N) and phosphoprotein (P). Sera from 15 of the 23 SSPE patients were reactive to MV-fusion protein (F). Faint reactivity was obtained against MV-matrix (M) or hemagglutinin protein (H) in the minority of samples (40 and 20%, respectively). CSF samples of MS patients only revealed a clear response to N, and in two cases to F. The other proteins were not recognized in the CSF samples of MS patients. In contrast to SSPE, the IEF of CSF from MS patients revealed only few MV-specific oligoclonal bands. In the CSF samples from APME patients, intrathecal MV antibodies were not detected. CONCLUSIONS: This study shows that discrimination between SSPE and MS can be achieved in doubtful cases by IEF using MV-N, P and F proteins.

[New aspects in the diagnosis of herpes simplex virus encephalitis--case reports and virologic findings]. / Neue Aspekte in der Diagnostik der Herpes-simplex-Virusenzephalitis--Fallberichte und virologische Befunde.

Herpes simplex virus (HSV) is one of the most common causes of severe necrotizing encephalitis in men. Without adequate acyclovir treatment it has a mortality rate of 70%. An early, reliable virological diagnosis is therefore essential. In this study, two newly-introduced diagnostic methods were assessed and compared: Polymerase Chain Reaction (PCR) for detection of HSV-genome in cerebrospinal fluid and HSV-specific isoelectric focusing (IEF) for detection of intrathecal virus specific antibody synthesis in HSV encephalitis (HSVE). PCR proved more effective for the early diagnosis of HSVE and HSVE relapses, whereas HSV-specific IEF was superior for the diagnosis of HSVE in post-acute stages of the disease.

The diagnostic significance of the polymerase chain reaction and isoelectric focusing in herpes simplex virus encephalitis.

In this study, serum and CSF samples of 55 neurological patients have been examined to confirm the diagnosis of herpes simplex virus encephalitis (HSVE). Different methods were applied, including serological titer evaluations, determination of intrathecally-produced HSV-specific antibodies by isoelectric focusing with affinity immunoblotting (IEF), as well as HSV-specific ELISA and HSV-specific polymerase chain reaction (PCR). The results of IEF and PCR have been compared and contrasted to develop general directions for virological diagnosis of HSVE. Of 14 patients suffering from clinically diagnosed HSVE, HSVE was confirmed in 12 cases by the demonstration of PCR or IEF positivity. A HSV-specific CNS infection could be excluded in 2 of these 14 patients. In 17 patients suffering from non-HSVE, PCR and IEF results were negative. Twenty-four patients, suffering from other neurological diseases, serving as a control group, were PCR- and HSV-IEF-negative. The study indicated that there are two possibilities for unequivocal demonstration of HSV-specific CNS involvement: first, performance of PCR especially in the acute phase of disease and in suspicious relapses, and second, performance of HSV-specific IEF for determination of intrathecally synthesized HSV-specific antibodies. It is suggested that these two methods should be introduced in routine diagnosis of viral encephalitis.

[Subclinical course of Epstein-Barr virus infection in infants with transient hepatitis]. / Subklinischer Verlauf einer Epstein-Barr-Virusinfektion im Säuglingsalter mit transienter Hepatitis.

Epstein-Barr-Virus-(EBV)-infections are well known to cause acute infectious mononucleosis in adolescents and young adults. However in early infancy and childhood these infections are usually mild and clinically inapparent. In the following we describe a case of a primary EBV-infection in infancy, which led to a transient hepatitis, identified by pathologically elevated liver enzymes.