A Crimean-Congo hemorrhagic fever (CCHF) viral vaccine expressing nucleoprotein is immunogenic but fails to confer protection against lethal disease.

Crimean-Congo Hemorrhagic Fever (CCHF) is a severe tick-borne disease, endemic in many countries in Africa, the Middle East, Eastern Europe and Asia. Between 15-70% of reported cases are fatal with no approved vaccine available. In the present study, the attenuated poxvirus vector, Modified Vaccinia virus Ankara, was used to develop a recombinant candidate vaccine expressing the CCHF virus nucleoprotein. Cellular and humoral immunogenicity was confirmed in 2 mouse strains, including type I interferon receptor knockout mice, which are susceptible to CCHF disease. Despite the immune responses generated post-immunisation, the vaccine failed to protect animals from lethal disease in a challenge model.

Effective binding of a phosphatidylserine-targeting antibody to Ebola virus infected cells and purified virions.

Ebola virus is responsible for causing severe hemorrhagic fevers, with case fatality rates of up to 90%. Currently, no antiviral or vaccine is licensed against Ebola virus. A phosphatidylserine-targeting antibody (PGN401, bavituximab) has previously been shown to have broad-spectrum antiviral activity. Here, we demonstrate that PGN401 specifically binds to Ebola virus and recognizes infected cells. Our study provides the first evidence of phosphatidylserine-targeting antibody reactivity against Ebola virus.

Sequence of pathogenic events in cynomolgus macaques infected with aerosolized monkeypox virus.

UNLABELLED: To evaluate new vaccines when human efficacy studies are not possible, the FDA's "Animal Rule" requires well-characterized models of infection. Thus, in the present study, the early pathogenic events of monkeypox infection in nonhuman primates, a surrogate for variola virus infection, were characterized. Cynomolgus macaques were exposed to aerosolized monkeypox virus (10(5) PFU). Clinical observations, viral loads, immune responses, and pathological changes were examined on days 2, 4, 6, 8, 10, and 12 postchallenge. Viral DNA (vDNA) was detected in the lungs on day 2 postchallenge, and viral antigen was detected, by immunostaining, in the epithelium of bronchi, bronchioles, and alveolar walls. Lesions comprised rare foci of dysplastic and sloughed cells in respiratory bronchioles. By day 4, vDNA was detected in the throat, tonsil, and spleen, and monkeypox antigen was detected in the lung, hilar and submandibular lymph nodes, spleen, and colon. Lung lesions comprised focal epithelial necrosis and inflammation. Body temperature peaked on day 6, pox lesions appeared on the skin, and lesions, with positive immunostaining, were present in the lung, tonsil, spleen, lymph nodes, and colon. By day 8, vDNA was present in 9/13 tissues. Blood concentrations of interleukin 1ra (IL-1ra), IL-6, and gamma interferon (IFN-γ) increased markedly. By day 10, circulating IgG antibody concentrations increased, and on day 12, animals showed early signs of recovery. These results define early events occurring in an inhalational macaque monkeypox infection model, supporting its use as a surrogate model for human smallpox. IMPORTANCE: Bioterrorism poses a major threat to public health, as the deliberate release of infectious agents, such smallpox or a related virus, monkeypox, would have catastrophic consequences. The development and testing of new medical countermeasures, e.g., vaccines, are thus priorities; however, tests for efficacy in humans cannot be performed because it would be unethical and field trials are not feasible. To overcome this, the FDA may grant marketing approval of a new product based upon the "Animal Rule," in which interventions are tested for efficacy in well-characterized animal models. Monkeypox virus infection of nonhuman primates (NHPs) presents a potential surrogate disease model for smallpox. Previously, the later stages of monkeypox infection were defined, but the early course of infection remains unstudied. Here, the early pathogenic events of inhalational monkeypox infection in NHPs were characterized, and the results support the use of this surrogate model for testing human smallpox interventions.

Efficacy of primate humoral passive transfer in a murine model of pneumonic plague is mouse strain-dependent.

New vaccines against biodefense-related and emerging pathogens are being prepared for licensure using the US Federal Drug Administration's "Animal Rule." This allows licensure of drugs and vaccines using protection data generated in animal models. A new acellular plague vaccine composed of two separate recombinant proteins (rF1 and rV) has been developed and assessed for immunogenicity in humans. Using serum obtained from human volunteers immunised with various doses of this vaccine and from immunised cynomolgus macaques, we assessed the pharmacokinetic properties of human and cynomolgus macaque IgG in BALB/c and the NIH Swiss derived Hsd:NIHS mice, respectively. Using human and cynomolgus macaque serum with known ELISA antibody titres against both vaccine components, we have shown that passive immunisation of human and nonhuman primate serum provides a reproducible delay in median time to death in mice exposed to a lethal aerosol of plague. In addition, we have shown that Hsd:NIHS mice are a better model for humoral passive transfer studies than BALB/c mice.

Development of an indirect ELISA method for the parallel measurement of IgG and IgM antibodies against Crimean-Congo haemorrhagic fever (CCHF) virus using recombinant nucleoprotein as antigen.

Recombinant nucleoprotein from Crimean-Congo Haemorrhagic Fever (CCHF) virus was successfully derived from a baculovirus expression system and purified for use in a novel enzyme-linked immunosorbent assay (ELISA) diagnostic test. Comparable tests were used for detection of IgG and IgM antibodies, thus allowing efficient detection of both antibodies in parallel. The major benefits of the assay also included removing any requirement for polyclonal sera, thus eliminating variation in preparations and allowing standardisation between laboratories. The assay was successfully tested using a panel of positive sera supplied from samples identified as being positive in Turkey, Tajikistan and Kosovo and shown to be sensitive and specific. It is envisaged that this simple diagnostic ELISA for CCHF virus infection which removes the reliance on polyclonal antibody preparations, will be accessible to a wider range of laboratories enabling them to carry out routine diagnosis. This will improve the efficiency of diagnosis and subsequent management of infected patients.

Filling the gap: weaning practices of children aged 0-2 years in western metropolitan Melbourne.

OBJECTIVE: To investigate weaning practices of children aged 0-2 years in Melbourne's western metropolitan region (WMR). METHODOLOGY: The study group comprised children aged 0-2 years attending six maternal and child health centres in the City of Brimbank in Melbourne's WMR. One hundred and nineteen parents were asked to complete a questionnaire about their child. Interpreters were used as required. In total, 115 questionnaires were completed representing a response rate of 97%. All maternal and child health nurses working in the target region were asked to complete a questionnaire. Of these, five were selected randomly to participate in follow-up indepth interviews. Weaning was defined as the addition of food/fluids other than breast milk or formula. RESULTS: The mean age for the introduction of solids was 4.3 months. Of the children already introduced to solids, 67% had commenced between 4 and 6 months whilst 29% of the infants were taking solids by 3 months of age. Of the children already having juice, 85% had done so by the age of 6 months. The mean age for the introduction of cow's milk as the main drink was 10.7 months (S.D. 2.8). For English speaking families the mean age was 11.3 months (S.D. 1.8) whilst for families speaking languages other than English the mean age was 9.7 months (S.D. 4.1). CONCLUSIONS: Most parents in this group were following the recommended guidelines for weaning. Some parents had poor knowledge of appropriate weaning time, use of fruit juice and introduction of cow's milk as the main drink. Additional information and education is required in these areas. Further investigation is required in relation to use of cow's milk and ethnicity.

The effect of lodoxamide tromethamine on the immediate asthmatic response to allergen provocation.

The inhibition of immediate allergen-induced airflow obstruction by lodoxamide tromethamine (LT), a new drug with properties considered to be similar to those of sodium cromoglycate, was studied in twelve young asthmatic volunteers. Single aerosolized doses of 0 X 01 mg LT, 0 X 1 mg LT or placebo were administered by inhalation 15 min prior to allergen provocation at weekly intervals, in a double-blind random order study. Following inhalation of both doses of LT a significantly greater amount of allergen had to be administered to cause a 20% fall in the forced expiratory volume in one second (FEV1) from control levels than was the case following placebo pre-treatment (P less than 0 X 001). After single-dose inhalation of LT only minor unwanted effects were recorded; in particular a transient feeling of heat in the upper respiratory tract after inhalation of the higher dose of drug.

Routine antibiotics in hospital management of acute asthma.

The value of antibiotics was assessed in a randomised, double-blind study of amoxycillin and placebo in sixty adults admitted to hospital with acute exacerbations of asthma. 37 exacerbations were treated with amoxycillin and 34 were treated with placebo. Response to treatment was closely monitored but no significant difference in improvement was demonstrated between groups for length of hospital stay, time taken for 50% improvement in symptoms, patient's self assessment and respiratory function, and symptoms and respiratory function at time of discharge from hospital. Antibiotics should not be given routinely to patients admitted to hospital with acute exacerbations of asthma.

Respiratory tract reactions to western red cedar.

1 Asthma due to western red cedar (Thuja plicata) is well recognized, but has not been described frequently in the UK. Two patients who developed asthma and rhinitis due to occupational contact with western red cedar were studied. Both patients developed late asthmatic responses following bronchial challenge with western red cedar. 2 The challenge technique and the results of comparison between different wood dusts and dust extract are described. 3 The technique of anterior rhinometry was used to follow the nasal response to challenge in one patient, and demonstrated a late nasal reaction which followed a similar time course to the bronchial response.