Medical management of biological warfare and bioterrorism: place of the immunoprevention and the immunotherapy.

Biological weapons are considered as mass destruction and terror weapons. Terrorism including bioterrorism is the major threat in the future conflicts for our nations. The aim of bioterrorism is more related to the potential disorganisation of the society than to the lethal effects of the agents used. The dramatic consequences cannot be discarded, especially if contagious agents such viral are used. The preparation of specific defence measures is a major challenge for our countries. The knowledge acquired from the struggle against natural infectious diseases and recent events are essential to improve behaviours to face the biological weapon threats. The defence attitude is based on the anticipation of the threat, the management of the victims, and the restoration of the operational capabilities. This global defence attitude implies six important functions: (i) alert, (ii) detection and diagnosis, (iii) availability of pharmaceutical countermeasures such as vaccine, sera and anti-infectious medicine and products, (iv) medical management of victims, (v) training and information, (vi) research and development. Passive and active immunoprevention and immuntherapy belong to the approaches discussed in the context of bioterrorism countermeasures. Further researches might be focused on these topics.

Interferon, ribavirin, 6-azauridine and glycyrrhizin: antiviral compounds active against pathogenic flaviviruses.

Ribavirin, interferon-alpha (IFN-alpha), 6-azauridine and glycyrrhizin were tested in vitro for their antiviral activities against 11 pathogenic flaviviruses belonging to principal antigenic complexes or individual serogroups of medical importance: dengue, Japanese encephalitis, mammalian tick-borne and yellow fever virus (YFV) groups. Antiviral activity was estimated by the reduction of the cytopathic effect of each flavivirus in Vero cells and by the reduction in virus titer. Cytotoxicity was evaluated by determining the inhibition of Trypan blue exclusion in confluent cell cultures and by the evaluation of the inhibitory effect on cell growth. The specificity of action of each tested compound was estimated by the selectivity index (CC(50)/EC(50)). IFN-alpha proved to be a selective and potent inhibitor of the replication of the 11 tested pathogenic flaviviruses. Ribavirin and 6-azauridine proved to be active on the replication of the 11 tested pathogenic flaviviruses at the concentrations which did not alter normal cell morphology, but they were not selective inhibitors when selectivity indices were evaluated with regard to the inhibition of cell growth because of their cytostatic effect. Glycyrrhizin inhibited the replication of flaviviruses at high non-cytotoxic concentrations. These antiflavivirus compounds should be further evaluated for their efficacy in the treatment of flavivirus infections in vivo.

Fever-like thermal conditions regulate the activation of maturing dendritic cells.

Fever is one of the most frequent clinical signs encountered in pathology, especially with respect to infectious diseases. It is currently thought that the role of fever on immunity is limited to activation of innate immunity; however, its relevance to activation of adaptive immunity remains unclear. Dendritic cells (DCs) that behave as sentinels of the immune system provide an important bridge between innate and adaptive immunity. To highlight the role of fever on adaptive immunity, we exposed murine bone marrow-derived lipopolysaccharide (LPS)- or live bacteria-maturing DCs over a 3-h period to 37 degrees C or to fever-like thermal conditions (39 degrees C or 40 degrees C). At these three temperatures, we measured the kinetics of cytokine production and the ability of DCs to induce an allogeneic mixed lymphocyte reaction. Our results show that short exposure of DCs to temperatures of 39 degrees C or 40 degrees C differentially increased the secretion of interleukin (IL)-12p70 and decreased the secretion of IL-10 and tumor necrosis factor alpha by maturing DCs. These fever-like conditions induced a regulation of cytokine production at the single-cell level. In addition, short-term exposed LPS-maturing DCs to 39 degrees C induced a stronger reaction with allogeneic CD4(+) T cells than maturing DCs incubated at 37 degrees C. These results provide evidence that temperature regulates cytokine secretion and DC functions, both of which are of particular importance in bacterial diseases.

Polystyrene derivatives substituted with arginine interact with Babanki (Togaviridae) and Kedougou (Flaviviridae) viruses.

Outbreaks of new or old diseases appear primarily in tropical zones such as Africa, south and central America, or Asia. Among these diseases, those induced by Arboviruses (the best known of which are being yellow fever, dengue, Ebola, and Sindbis) are under intensive observation by the World Health Organization. Rapid isolation and identification of the viral species is the first step in the diagnosis, study, and control of epidemics. One major problem with the isolation of viruses is capturing sufficient numbers of viral particles to test. The work presented in this report addresses this question. We have tested the interaction between Babanki (Togaviridae), Kedougou (Flaviviridae) viruses, and a range of insoluble polystyrene derivatives substituted with arginine groups. Insoluble functionalized copolymers were found to develop specific interactions with viruses through chemical groups present on their surfaces. The adsorption of viruses varied according to the percentage of arginine substituted onto the polymer, with a maximum value for both viruses of about 20% of grafting rate. It was also found that the Kedougou virus displayed the highest affinity for this polymer.

NS3 protease of Langat tick-borne flavivirus cleaves serine protease substrates.

Langat (LGT) virus, initially isolated in 1956 from ticks in Malaysia, is a naturally occurring nonpathogenic virus with a very close antigenicity to the highly pathogenic tick-borne encephalitis (TBE) Western subtype virus and TBE Far Eastern subtype virus. NS3, the second largest viral protein of LGT virus, is highly conserved among flaviviruses and contains a characteristic protease moiety (NS3 pro). NS3 pro represents an attractive target for anti-protease molecules against TBE virus. We report herein a purification method specially designed for NS3 pro of LGT using a strategy for proper refolding coupled with the enzymatic characterisation of the protein. Different p-nitroanilide substrates, defined on canonic sequences for their susceptibility to Ser-protease, were applied to the proteolytic assays of the protein. The highest values were obtained from substrates containing an Arg or Lys (amino acid) residue at the P1 position. This purification method will facilitate the future development of reliable testing procedures for anti-proteases directed to NS3 proteins.

Mapping of a dengue virus neutralizing epitope critical for the infectivity of all serotypes: insight into the neutralization mechanism.

Dengue virus infections are a growing public health concern and strategies to control the spread of the virus are urgently needed. The murine monoclonal antibody 4E11 might be of interest, since it neutralizes dengue viruses of all serotypes by binding to the 296-400 segment of the major dengue virus envelope glycoprotein (DE). When phage-displayed peptide libraries were screened by affinity for 4E11, phage clone C1 was selected with a 50% frequency. C1 shared three of nine residues with DE(306-314) and showed significant reactivity to 4E11 in ELISA. C1-induced antibodies cross-reacted with DE(296-400) in mice, suggesting that it was a structural equivalent of the native epitope of 4E11 on DE. Accordingly, 4E11 bound to the DE(306-314) synthetic peptide and this reaction was inhibited by DE(296-400). Moreover, DE(306-314) could block dengue virus infection of target cells in an in vitro assay. A three-dimensional model of DE revealed that the three amino acids shared by DE(296-400) and C1 were exposed to the solvent and suggested that most of the amino acids comprising the 4E11 epitope were located in the DE(306-314) region. Since 4E11 blocked the binding of DE(296-400) to heparin, which is a highly sulfated heparan sulfate (HSHS) molecule, 4E11 may act by neutralizing the interaction of DE(306-314) with target cell-displayed HSHS. Our data suggest that the DE(306-314) segment is critical for the infectivity of all dengue virus serotypes and that molecules that block the binding of DE(306-314) to HSHS may be antiviral reagents of therapeutic interest.