Functional genomic and serological analysis of the protective immune response resulting from vaccination of macaques with an NS1-truncated influenza virus.

We are still inadequately prepared for an influenza pandemic due to the lack of a vaccine effective for subtypes to which the majority of the human population has no prior immunity and which could be produced rapidly in sufficient quantities. There is therefore an urgent need to investigate novel vaccination approaches. Using a combination of genomic and traditional tools, this study compares the protective efficacy in macaques of an intrarespiratory live influenza virus vaccine produced by truncating NS1 in the human influenza A/Texas/36/91 (H1N1) virus with that of a conventional vaccine based on formalin-killed whole virus. After homologous challenge, animals in the live-vaccine group had greatly reduced viral replication and pathology in lungs and reduced upper respiratory inflammation. They also had lesser induction of innate immune pathways in lungs and of interferon-sensitive genes in bronchial epithelium. This postchallenge response contrasted with that shortly after vaccination, when more expression of interferon-sensitive genes was observed in bronchial cells from the live-vaccine group. This suggested induction of a strong innate immune response shortly after vaccination with the NS1-truncated virus, followed by greater maturity of the postchallenge immune response, as demonstrated with robust influenza virus-specific CD4+ T-cell proliferation, immunoglobulin G production, and transcriptional induction of T- and B-cell pathways in lung tissue. In conclusion, a single respiratory tract inoculation with an NS1-truncated influenza virus was effective in protecting nonhuman primates from homologous challenge. This protection was achieved in the absence of significant or long-lasting adverse effects and through induction of a robust adaptive immune response.

Integrated molecular signature of disease: analysis of influenza virus-infected macaques through functional genomics and proteomics.

Recent outbreaks of avian influenza in humans have stressed the need for an improved nonhuman primate model of influenza pathogenesis. In order to further develop a macaque model, we expanded our previous in vivo genomics experiments with influenza virus-infected macaques by focusing on the innate immune response at day 2 postinoculation and on gene expression in affected lung tissue with viral genetic material present. Finally, we sought to identify signature genes for early infection in whole blood. For these purposes, we infected six pigtailed macaques (Macaca nemestrina) with reconstructed influenza A/Texas/36/91 virus and three control animals with a sham inoculate. We sacrificed one control and two experimental animals at days 2, 4, and 7 postinfection. Lung tissue was harvested for pathology, gene expression profiling, and proteomics. Blood was collected for genomics every other day from each animal until the experimental endpoint. Gross and microscopic pathology, immunohistochemistry, viral gene expression by arrays, and/or quantitative real-time reverse transcription-PCR confirmed successful yet mild infections in all experimental animals. Genomic experiments were performed using macaque-specific oligonucleotide arrays, and high-throughput proteomics revealed the host response to infection at the mRNA and protein levels. Our data showed dramatic differences in gene expression within regions in influenza virus-induced lesions based on the presence or absence of viral mRNA. We also identified genes tightly coregulated in peripheral white blood cells and in lung tissue at day 2 postinoculation. This latter finding opens the possibility of using gene expression arrays on whole blood to detect infection after exposure but prior to onset of symptoms or shedding.

Comparison of gastrointestinal tolerability and patient preference for treatment with the 625 mg and 250 mg nelfinavir tablet formulations.

OBJECTIVES: To compare gastrointestinal (GI) tolerability and patient preference for the new 625 mg formulation of nelfinavir (NFV) and the marketed 250 mg tablets (Viracept) in HIV-1-infected patients. METHODS: Virologically controlled patients (n=126) treated with a nelfinavir (NFV) 250 mg-containing regimen for > or =8 weeks completed a stool diary for 14 days to assess baseline bowel function. After switching to the NFV 625 mg formulation [1250 mg twice a day (bid)] for 28 days, patients continued their stool diaries and at study completion answered a questionnaire regarding formulation preferences. RESULTS: The incidence and mean weekly duration of GI upset over a 2-week period were lower with NFV 625 mg than with NFV 250 mg (79.8% vs. 84.9% of patients and 2.1 vs. 3.0 days, respectively). Fewer patients experienced moderate or severe diarrhoea with NFV 625 mg (6.5% vs. 11.1%), and the incidence of investigator-assessed diarrhoea also decreased with NFV 625 mg. Importantly, there was a significant improvement overall in the incidence of diarrhoea (any grade) when patients switched to NFV 625 mg [38 of 124 (31%) improving, 69 of 124 (56%) stable and 17 of 124 (14%) worsening on NFV 625 mg; P<0.01]. At study completion, most patients expressed a preference to continue treatment with NFV 625 mg [112 of 122 (91.8%); P<0.0001], with only one patient (0.8%) preferring to resume treatment with NFV 250 mg. The new formulation was well tolerated with no new safety concerns. CONCLUSIONS: The new NFV 625 mg formulation is better tolerated and preferred by patients switching from NFV 250 mg tablets. By reducing the daily pill count and improving GI tolerability, the NFV 625 mg formulation may enhance patient adherence to NFV-containing antiretroviral regimens and thus potentially improve virological outcomes.

[The risk of contracting hepatitis A or hepatitis B run by visitors to the Mediterranean and Eastern Europe]. / Infektionsrisiko für Reisende im Mittelmeerraum und in Osteuropa. Hepatitis A und B: Souvenirs, die keiner haben will.

Hepatitis A and B continue to represent a risk for travelers. Those traveling to tropical countries are usually aware of this risk, while those visiting a Mediterranean country often are not. This investigation presents a destination-related risk assessment and vaccination recommendations for travellers, on the basis of incidences and prevalences of hepatitis A and hepatitis B. For hepatitis B, the WHO classification based on HBsAg prevalence has been selected. For hepatitis A, countries have been categorized as low-, moderate- or high-risk. A comparison of infection risk with the numbers of airline passengers showed that in particular the large number of travelers to Tunisia, Turkey and Egypt, in conjunction with hygienic conditions in those countries, represent a source of imported hepatitis A. With regard to destinations in the Mediterranean and Eastern Europe, too, the risk of contracting hepatitis A or hepatitis B is not always negligible, so that vaccination may need to be recommended.

Sublingual specific immunotherapy for adults and children: a post-marketing surveillance study.

BACKGROUND: Sublingual immunotherapy (SLIT) is a relatively new form of treatment for type I allergies and a good safety profile is rapidly being established. Evidence on the efficacy of SLIT is increasing, and the present study provides further supportive data. We describe the results of treatment with a SLIT vaccine formulated with a range of allergen extracts obtained by allergologists in daily clinical practice. METHODS: Adult and child patients (n = 159, 81 males, 78 females) with confirmed type I allergic sensitivities were treated with a standardized SLIT vaccine (ORALVAC) using the sublingual-swallow method. Evaluation of the efficacy of SLIT was based on the consumption of anti-allergic medication and a global assessment. Tolerability assessment was based on the incidence of local or systemic reactions. RESULTS: Medication use was significantly reduced compared with that in previous years (p = 0.023). In a large subgroup of patients treated for pollen sensitivity the significance was stronger (p = 0.016). Global assessment revealed that only 3.5 % of patients showed no change in symptoms after therapy. High tolerability was achieved and no serious or severe adverse effects were observed. CONCLUSION: Over a one-year period, adult and child patients with a variety of type I allergies were treated with a SLIT vaccine that has shown significant efficacy and was well-tolerated with no serious or severe adverse events.

Single-course specific immunotherapy with mixed pollen allergoids: results of a multi-centre study.

BACKGROUND: A short-term immunotherapy vaccine for the treatment of pollen allergy has been developed utilising L-tyrosine adsorbed allergoids. The reduced number of injections could provide advantages over long-term therapy schedules. This would improve compliance and support application of specific immunotherapy (SIT) to a greater extent. We report a multicenter study to evaluate the efficacy and safety of this treatment in a clinical practice setting. METHODS: Patients (n = 1808) with a diagnosis of sensitivities to various pollens and symptoms of allergic asthma and/or allergic rhinitis and/or allergic conjunctivitis were selected. The vaccine formulation was made up according to individual sensitivities and contained L-tyrosine adsorbed allergoids. The patients were treated with a 3-injection initial course followed by a 3-injection maintenance course. Efficacy was measured by consumption of symptomatic anti-allergic medication compared with that in the previous season and by physician assessment using a 5-point scale. All adverse events were recorded. RESULTS: Efficacy was demonstrated by a considerable decrease in regular and frequent use of medication compared with that in the previous season (p < 0.001). In addition, in 80 % of the patients, the physician's assessment was either "good" or "very good". These outcomes were unaffected by the closeness of the treatment course to the onset of the pollen season. Tolerability was good and most local and systemic reactions were mild. CONCLUSIONS: The treatment of pollen-allergic patients with a short-term SIT using a 6-injection pollen allergoid/L-tyrosine vaccine in a clinical practice setting provided a high level of efficacy with a low incidence of mainly mild adverse events.

Human cutaneous leishmaniasis caused by Leishmania naiffi is wide-spread in South America.

Four human cases of localized cutaneous leishmaniasis caused by Leishmania naiffi are reported. Two of the cases were infected in French Guiana, one in French Guiana or Martinique, and the other in Ecuador or Peru. The geographical distribution of L. naiffi is clearly larger than that initially reported. Three zymodemes were represented by the four isolates, confirming that there is intraspecific polymorphism in L. naiffi.

Bone dysplasia sclerosteosis results from loss of the SOST gene product, a novel cystine knot-containing protein.

Sclerosteosis is an autosomal recessive sclerosing bone dysplasia characterized by progressive skeletal overgrowth. The majority of affected individuals have been reported in the Afrikaner population of South Africa, where a high incidence of the disorder occurs as a result of a founder effect. Homozygosity mapping in Afrikaner families along with analysis of historical recombinants localized sclerosteosis to an interval of approximately 2 cM between the loci D17S1787 and D17S930 on chromosome 17q12-q21. Here we report two independent mutations in a novel gene, termed "SOST." Affected Afrikaners carry a nonsense mutation near the amino terminus of the encoded protein, whereas an unrelated affected person of Senegalese origin carries a splicing mutation within the single intron of the gene. The SOST gene encodes a protein that shares similarity with a class of cystine knot-containing factors including dan, cerberus, gremlin, prdc, and caronte. The specific and progressive effect on bone formation observed in individuals affected with sclerosteosis, along with the data presented in this study, together suggest that the SOST gene encodes an important new regulator of bone homeostasis.

X-linked neonatal diabetes mellitus, enteropathy and endocrinopathy syndrome is the human equivalent of mouse scurfy.

To determine whether human X-linked neonatal diabetes mellitus, enteropathy and endocrinopathy syndrome (IPEX; MIM 304930) is the genetic equivalent of the scurfy (sf) mouse, we sequenced the human ortholog (FOXP3) of the gene mutated in scurfy mice (Foxp3), in IPEX patients. We found four non-polymorphic mutations. Each mutation affects the forkhead/winged-helix domain of the scurfin protein, indicating that the mutations may disrupt critical DNA interactions.

Influence of rheumatoid factor on the specificity of a rapid immunochromatographic test for diagnosing dengue infection.

In order to determine the influence of rheumatoid factor in serum specimens on the specificity of an immunochromatographic test for the detection of antibodies against dengue virus, 50 clinical specimens containing rheumatoid factor were evaluated for cross reactivity with a commercially available assay. While specificity for the detection of immunoglobulin (Ig) G antibodies was 100%, the IgM component of the test showed false-positive results in 26% of cases. Thus, it is important to consider interference by rheumatoid factor when using immunochromatographic assays for the detection of specific IgM produced during dengue infection.

Population structure of recrudescent Plasmodium falciparum isolates from western Uganda.

It has been proposed that polymorphisms of the Merozoite Surface Protein 1 and 2 (MSP1 and MSP2) and the Glutamate Rich Protein (GLURP) genes can be considered as genetic markers for the genotyping of field populations of Plasmodium falciparum. During a field study on in vivo drug resistance against chloroquine, sulphadoxine/pyrimethamine (S/P) and cotrimoxazole in West Uganda, sensitive and resistant isolates were collected from patients by fingerprick for genotyping. 59 (72.8%) of the 81 P. falciparum samples isolated at day 0 showed multiclonal infection with 2-7 clones. Among the isolates we investigated, presence of the allelic family MAD20 of MSP1 at day 0 was significantly (P = 0.0041) associated with decreased resistance to antimalarials. Use of this method in a field study on in vivo drug resistance demonstrates another potential application of genotyping as a tool for epidemiological investigations.

[Persistent arthralgias in Ross-River-Virus disease after travel to the South Pacific]. / Persistierende Arthralgien bei Ross-River-Virus-Erkrankung nach Ozeanien-Reise.

HISTORY AND CLINICAL FINDINGS: A 57-year-old patient presented with malaise and severe persistent arthralgia of the left shoulder. He reported an acute illness with fever, generalized myalgia and arthralgias of the large joints which had started one month earlier during his flight back to Germany after a two weeks trip to the South Pacific. Physical examination showed extensive pain on palpation of the glenohumeral and acromioclavicular joints with impairment of active and passive mobility. Investigation of the cervical spine was normal. INVESTIGATIONS: Apart from elevated C-reactive protein and erythrocyte sedimentation rate levels, routine laboratory investigations were normal including negative immunodiagnostic tests for autoantibodies and various global infections that may be associated with arthritis. Immunofluorescence tests showed significant levels of specific IgM- and IgG-antibodies against Ross River virus (RRV) but not against other arboviruses endemic in the South Pacific and Australia (Dengue, West Nile, Chikungunya, Sindbis, Barmah Forest). This was confirmed by a positive RRV neutralisation test. Attempts at virus isolation and detection of viral RNA by PCR were not successful. TREATMENT AND COURSE: Symptomatic treatment with high doses of diclofenac quickly led to pain relief, and arthralgias receded within 10 days after begin of treatment. However, several bouts of arthralgia of the left shoulder and left knee occurred during a period of 4 months. CONCLUSIONS: Because of the current epidemiological situation in the South Pacific and Australia, infections by arboviruses like RRV should be considered in travellers returning from these areas with severe and persistent arthralgia of unknown origin, even in the absence of fever and other symptoms of acute infection.

Geographic differences in the sensitivity of a polymerase chain reaction for the detection of Plasmodium falciparum infection.

The amplification of target DNA by highly specific probes using the polymerase chain reaction (PCR) provides a highly sensitive and specific method for the detection of malaria infection. The use the of PCR in settings with varying endemicity within one survey area has not been investigated intensively. Therefore, a cross-sectional study was conducted in the districts of Kabarole and Bundibugyo in western Uganda using material from three villages with different epidemiologic situations regarding malaria and DNA primers for a PCR that had shown satisfactory sensitivity and specificity in previous trials. The sensitivity of the PCR varied significantly (P < 0.001) in the three survey villages (between 63.2% and 83.9% for the primer pair K1-14-1 and between 37.9% and 69.9% for the primer pair MSP-1) and was highly linked to geographic differences and social exchanges of the inhabitants with other areas of the district. According to the results of this investigation, it is advisable not to use a single primer pair in epidemiologic field studies for the detection of falciparum malaria. The use of combined primer pairs and the frequent confirmation of the results by microscopy are recommended.

Plasmodium falciparum and Plasmodium berghei: effect of magnesium on the development of parasitemia.

The in vitro growth of Plasmodium falciparum was reduced by 35 and 43% through high concentrations (5 mmole/liter) of magnesium in RPMI medium and magnesium-free medium, respectively, after 48 hr, whereas no significant inhibition could be observed under these conditions after 24 hr cultivation in the respective medium. Levels of magnesium between 0.5 and 3 mmole/liter showed no inhibitory effect on the in vitro growth of P. falciparum even after long-term exposure for 7 days. The 50 and 90% chloroquine inhibitory concentrations of the chloroquine-resistant strain K1 after 24 hr were reduced to some extent in the presence of magnesium at 5 mmole/liter, but less than in the presence of verapamil at 10 mumole/liter, which showed intrinsic activities at this concentration and which completely reversed resistance. However, high physiologic magnesium plasma levels were associated with a significantly longer survival time of NMRI mice infected with P. berghei strain ANKA, compared to normal physiological plasma magnesium levels. It is concluded that in the case of clinically symptomatic magnesium deficiency, supplementation of magnesium will not aggravate concomitant plasmodial infections and therefore should not be withheld.

[Chronic subdural hematoma in aged patients. Diagnostic problems]. / Chronisches Subduralhämatom beim Alterspatienten. Zur Problematik der Diagnose.

The history, clinical features, laboratory tests and admission circumstances were analysed retrospectively in 14 patients, aged 60 to 87 years, who were found to have a chronic subdural haematoma. Typical clinical features consisted of dominant psychiatric disorders, slow progression and frequently only traces of neurological signs. In only four patients was headache the initial symptom. Delay in diagnosis and treatment worsened the prognosis. Several factors account for the nonspecific signs and symptoms: diffuse cerebral atrophy, dementia of a degenerative or vascular nature, and cerebral decompensation. Computed tomography immediately after a trauma may be negative.