Lymphocytic choriomeningitis virus meningitis after needlestick injury: a case report.

Background: Needlestick accidents while handling of infectious material in research laboratories can lead to life-threatening infections in laboratory personnel. In laboratories working with the lymphocytic choriomeningitis virus (LCMV), the virus can be transmitted to humans through needlestick injury and lead to serious acute illness up to meningitis. Case presentation: We report of a case of LCMV meningitis in a laboratory worker who sustained a penetrating needlestick injury with a LCMV-contaminated hollow needle whilst disposing of a used syringe into the sharps waste bin. Four days after needlestick injury the laboratory worker developed a systemic disease: 11 days after exposure, she was diagnosed with meningitis with clinical signs and symptoms of meningismus, photophobia, nausea and vomiting, requiring hospitalisation. The PCR was positive for LCMV from the blood sample. 18 days after exposure, seroconversion confirmed the diagnosis of LCMV-induced meningitis with an increase in specific LCMV-IgM antibodies to 1:10'240 (day 42: 1:20'480). Ten weeks after exposure, a follow-up titre for IgM returned negative, whereas IgG titre increased to 1:20'480. Conclusions: This is the first case report of a PCR-documented LCMV meningitis, coupled with seroconversion, following needlestick injury. It highlights the importance of infection prevention practices that comprise particularly well established safety precaution protocols in research laboratories handling this pathogenic virus, because exposure to even a small amount of LCMV can lead to a severe, life-threatening infection.

Biological roles and functional mechanisms of arenavirus Z protein in viral replication.

Arenaviruses can cause severe hemorrhagic fever diseases in humans, with limited prophylactic or therapeutic measures. A small RING-domain viral protein Z has been shown to mediate the formation of virus-like particles and to inhibit viral RNA synthesis, although its biological roles in an infectious viral life cycle have not been directly addressed. By taking advantage of the available reverse genetics system for a model arenavirus, Pichinde virus (PICV), we provide the direct evidence for the essential biological roles of the Z protein's conserved residues, including the G2 myristylation site, the conserved C and H residues of RING domain, and the poorly characterized C-terminal L79 and P80 residues. Dicodon substitutions within the late (L) domain (PSAPPYEP) of the PICV Z protein, although producing viable mutant viruses, have significantly reduced virus growth, a finding suggestive of an important role for the intact L domain in viral replication. Further structure-function analyses of both PICV and Lassa fever virus Z proteins suggest that arenavirus Z proteins have similar molecular mechanisms in mediating their multiple functions, with some interesting variations, such as the role of the G2 residue in blocking viral RNA synthesis. In summary, our studies have characterized the biological roles of the Z protein in an infectious arenavirus system and have shed important light on the distinct functions of its domains in virus budding and viral RNA regulation, the knowledge of which may lead to the development of novel antiviral drugs.

Like-acetylglucosaminyltransferase (LARGE)-dependent modification of dystroglycan at Thr-317/319 is required for laminin binding and arenavirus infection.

α-dystroglycan is a highly O-glycosylated extracellular matrix receptor that is required for anchoring of the basement membrane to the cell surface and for the entry of Old World arenaviruses into cells. Like-acetylglucosaminyltransferase (LARGE) is a key molecule that binds to the N-terminal domain of α-dystroglycan and attaches ligand-binding moieties to phosphorylated O-mannose on α-dystroglycan. Here we show that the LARGE modification required for laminin- and virus-binding occurs on specific Thr residues located at the extreme N terminus of the mucin-like domain of α-dystroglycan. Deletion and mutation analyses demonstrate that the ligand-binding activity of α-dystroglycan is conferred primarily by LARGE modification at Thr-317 and -319, within the highly conserved first 18 amino acids of the mucin-like domain. The importance of these paired residues in laminin-binding and clustering activity on myoblasts and in arenavirus cell entry is confirmed by mutational analysis with full-length dystroglycan. We further demonstrate that a sequence of five amino acids, Thr(317)ProThr(319)ProVal, contains phosphorylated O-glycosylation and, when modified by LARGE is sufficient for laminin-binding. Because the N-terminal region adjacent to the paired Thr residues is removed during posttranslational maturation of dystroglycan, our results demonstrate that the ligand-binding activity resides at the extreme N terminus of mature α-dystroglycan and is crucial for α-dystroglycan to coordinate the assembly of extracellular matrix proteins and to bind arenaviruses on the cell surface.

The role of the vascular endothelium in arenavirus haemorrhagic fevers.

Viral haemorrhagic fevers (VHF) caused by arenaviruses are among the most devastating emerging human diseases. The most important pathogen among the arenaviruses is Lassa virus (LASV), the causative agent of Lassa fever that is endemic to West Africa. On the South American continent, the New World arenavirus Junin virus (JUNV), Machupo (MACV), Guanarito (GTOV), and Sabia virus (SABV) have emerged as causative agents of severe VHFs. Clinical and experimental studies on arenavirus VHF have revealed a crucial role of the endothelium in their pathogenesis. However, in contrast to other VHFs, haemorrhages are not a salient feature of Lassa fever and fatal cases do not show overt destruction of vascular tissue. The functional alteration of the vascular endothelium that precede shock and death in fatal Lassa fever may be due to more subtle direct or indirect effects of the virus on endothelial cells. Haemorrhagic disease manifestations and vascular involvement are more pronounced in the VHF caused by the South American haemorrhagic fever viruses. Recent studies on JUNV revealed perturbation of specific endothelial cell function, including expression of cell adhesion molecules, coagulation factors, and vasoactive mediators as a consequence of productive viral infection. These studies provided first possible links to some of the vascular abnormalities observed in patients, however, their relevance in vivo remains to be investigated.

Fibroblastic reticular cells and their role in viral hemorrhagic fevers.

Viral hemorrhagic fevers (VHFs) caused by Ebola, Marburg and Lassa viruses often manifest as multiple organ dysfunction and hemorrhagic shock with high mortality. These viruses target numerous cell types, including monocytes and dendritic cells, which are primary early targets that mediate critical pathogenetic processes. This review focuses on fibroblastic reticular cells (FRCs), another prevalent infected cell type that is known as a key regulator of circulatory and immune functions. Viral infection of FRCs could have debilitating effects in secondary lymphoid organs and various other tissues. FRCs may also contribute to the spread of these deadly viruses throughout the body. Here, we review the salient features of these VHFs and the biology of FRCs, emphasizing the potential role of these cells in VHFs and the rapid deterioration of immune and hemovascular sytems that are characteristic of such acute infections.

[Venezuelan hemorrhagic fever: eight years of observation]. / Fiebre hemorrágica venezolana: ocho años de observación.

Venezuelan hemorrhagic fever (VHF) is a severe disease characterised by fever, malaise, sore throat, followed by abdominal pain, diarrhea, a variety of hemorrhagic manifestations and convulsions. The arenavirus Guanarito is the causal agent and the virus natural reservoir is the rodent Zygodontomys brevauda (cane mouse). The disease affect agricultural male workers, between 14-54 years of age, mainly from Guanarito municipality of Portuguesa state and adjacent regions of Barinas State. Since VHF emergency in 1989 up till 1997, 220 cases have been reported with a fatality rate of 33%. Epidemiological informations suggest that VHF has a cyclic behaviour, with epidemic periods of high incidence, every 4-5 years. During the interepidemic periods few VHF cases are reported.

Arboviruses Pathogenic for man in Brazil

Os mais importantes aspectos clinicicos e ecoepidemiologicos e aspectos preventivos acerca das arboviroses associadas com doenca humana no Brasil sao discutidos.Trinta e seis arbovirus dentre os tipos presentemente isolados no Pais tem sido incriminados como causadores de doenca humana. Destes, cinco sao importantes em termos de saude publica pois estao associados com epidemias , sao os virus Dengue (DEN), Mayaro(MAY), Oropouche (ORO), Rocio (ROC) e Febre amarela (FA). DEN e ORO estao associados com doenca humana epidemica em areas urbanas enquanto MAY, ROC e FA especialmente em areas rurais. Basicamente, o virus ORO determina um quadro febril algumas vezes acmpanhado por meningite asseptica. MAY e DEN sao responsaveis por quadros exantematicos, sendo que DEN, nos ultimos anos tem sido associado com quadros de febre hemorragica, o que sabidamente e o mecanismos pelo qual o virus FA determinaa sua apresentacao clinica classica e o ROC esta associado com graves quadros de encefalite. Trinta e um outros arbovirus tem sido associados com doenca febril benigna em poucos e esporadicos casos. Afora DEN e os Arenavirus Flexal e Sabia ( nao sao arbovirus), todos os arbovirus envolvidos com doenca humana na AmazoniaBrasileira, sao mantidos em natureza atraves de um ciclo silvestre desenvolvido na floresta, onde diversas especies de insetos hematofagos e vertebrados silvestres atuam como vetores e hospedeiros, respectivamente.O virus DEN tem um ciclo urbano em que o mosquito Aedes aegypti e o vetor e o homem atua como hospedeiro. Os arenavirus sao transmitidos diretamente ao homen atraves de excretas de roedores que sao seus principais hospedeiros.Excetuando os cinco virus associados com epidemias que causam um grande impacto socio-economico, inclusive levando a morte, casos verificados com FA, DEN e ROC, o verdadeiro papel dessesvirus como agentes sistematicos de doencas humanas e ainda puco conhecido. Novos estudos sao necessarios para esclarecer aspectos ainda obscuros acerca da epidemiologia da maioria desses arbovirus

Fiebre hemorrágica Venozolana: ocho anos de observacion / Venezuelan hemorrhagic fever: eight of observation

La fiebre hemorrágica Venezolana (FHV) es una enfermedad severa, caracterizada por fiebre, malestar general, dolor de cabeza dolor de garganta, seguida por dolor abdominal, diarrea, una variedad de manifestaciones hemorrágicas y convulsiones. El agente causal es el arenavirus Guanarito, el cual es mantenido en la naturaleza por el roedor de la especie Zygodontomys brevicauda: ratón de la caña de azúcar. La enfermedad afecta principalmente a agricultores del sexo masculino, en edades comprendidas entre 14- 54 años, procedentes del Municipio Guanarito del estado Portuguesa y áreas adyacentes del Estado Barinas. Desde la emergencia de la FHV en 1989 hasta 1997, se han reportado 220 casos con una letalidad del 33 por ciento. Tiene un comportamiento epidemiológico cíclico, registrándose periodos epidémicos cada 4-5 años, e interepidémicos con un numero reducido de casos.

Arenavirus infection--Connecticut, 1994.

On August 20, 1994, the Connecticut Department of Public Health and Addiction Services received a report of a case of acute illness in a virologist suspected to be associated with Sabiá virus, a newly described arenavirus. This report preliminary findings from the case investigation.

Pathogenesis of Pichinde virus infection in strain 13 guinea pigs: an immunocytochemical, virologic, and clinical chemistry study.

Pichinde virus has been adapted to produce lethal infection of Strain 13 guinea pigs. Viral replication and presence of viral antigen in frozen tissues stained by immunofluorescence has been previously described. Further investigation into the pathogenesis of this disease has been hampered by the lack of a light microscopic method for correlating histologic lesions and the presence of Pichinde viral antigens. For this purpose, we developed a sensitive immunocytochemical technique for staining Pichinde viral antigens in formalin-fixed, paraffin-embedded tissue. Enhancement of the immunocytochemical staining with nickel chloride markedly improved detection of viral antigens. We examined frozen and formalin-fixed tissues from Strain 13 guinea pigs for viral antigens by light microscopy and immunocytochemistry at various intervals after infection with Pichinde virus. Progressive involvement of different tissues correlated with organ injury measured by serum biochemical abnormalities. Pichinde viral antigen was first detected in splenic macrophages five days after infection and their subsequent destruction facilitated persistent viremia. The inability to clear virus led to multiple organ infection and vascular involvement. Ensuing infections involved particularly the liver, spleen, adrenal glands, lungs, and intestines. Gastroenteritis developed, with extensive involvement of the muscularis mucosa throughout the gastrointestinal tract. Water and food intake decreased rapidly after day 8, leading to marked weight loss. Fatty changes of the liver suggested metabolic derangement that was further exacerbated terminally by adrenal infection and pulmonary impairment.

Evidence for the involvement of sulfidopeptide leukotrienes in the pathogenesis of Pichinde virus infection in strain 13 guinea pigs.

Strain 13 guinea pigs inoculated subcutaneously with Pichinde virus developed fever, lost body weight, decreased water and food consumption, and died at 14 +/- 0.6 days. After FPL-55712, a leukotriene D4 antagonist, was administered beginning on PID (post-inoculation day) 3 for 18 days, the magnitude of body weight loss decreased and food intake increased, despite a persistent fever. The treated guinea pigs also survived significantly longer than infected animals receiving placebo injection (21 vs 14 days). Using guinea pig ileum bioassay and radioimmunoassay, we detected significant levels of plasma leukotrienes in Pichinde virus-infected guinea pigs on PID 11 and possibly PID 14. These findings strongly suggest that leukotrienes play a role in the pathogenesis of arenavirus infection and may account in part for the observed cardiac depression, pulmonary edema, and eventual death.

Generation of memory cell-mediated immune responses after secondary infection of mice with pichinde virus.

Pichinde virus (PV), a member of the arenavirus group, was found to elicit strong cell-mediated immune responses in various strains of mice. After primary i.v. inoculation, augmentation of natural killer (NK) cell activity occurred and peaked 3 to 4 days after infection. The NK response was followed by a second peak of cytotoxic activity that was found to be H-2 restricted, virus specific, and mediated by Thy-1.2+, Lyt-2.2+ lymphocytes. This cytotoxic T lymphocyte (CTL) response peaked 7 days post infection. Neutralizing antibodies were not detectable after PV infection of the mice. In light of this, we investigated the generation and kinetics of secondary cell-mediated immune responses after reinjection of homologous virus in vivo. Slight but significant augmentation of NK activity was observed 1 day after secondary virus challenge. As in the primary response, effectors of this NK activity rapidly became sensitive to anti-Thy-1.2 and complement treatment. NK activity rapidly returned to background levels and was followed by an anamnestic CTL response that peaked 4 days after reinjection of the virus. Thus, cell-mediated immune responses appeared more rapidly after secondary challenge in vivo, and the temporal relationship between NK and CTL generation was maintained. Both secondary NK and CTL responses were generated in mice that had been pretreated with cyclophosphamide (CY), suggesting that memory cell-mediated immune responses can be reactivated in vivo without undergoing cell division. In contrast, treatment with CY before primary infection delayed the appearance of virus-induced NK activity and abrogated the generation of H-2-restricted virus-specific CTL. Rechallenge of these CY-treated NK-primed mice resulted in the rapid generation of a secondary NK response that was not followed by either a primary or secondary CTL response. The data suggest that cells mediating a nonspecific effector function may possess specific memory. We discuss our results with respect to possible NK-CTL relationships.