Lassa fever encephalopathy: clinical and laboratory findings.

Clinical and laboratory findings are reported in nine patients who developed acute encephalopathy during the course of Lassa fever. The encephalopathy manifested 3-17 days after disease onset with confusion, followed rapidly by tremor (seven patients), grand mal convulsions (seven), abnormal posturing (three) and coma (eight); focal neurological signs and evidence of raised intracranial pressure were not seen. Eight patients died, most commonly from respiratory arrest following a protracted fit. Development of encephalopathy did not correlate with the presence of virus in cerebrospinal fluid (CSF), nor with virus antibodies in CSF and/or serum; thus, neither direct cytopathic nor immune-mediated mechanisms seem to be involved in its pathogenesis.

[Experimental Lassa fever in hamadryas baboons]. / Eksperimental'naia likhoradka Lassa u pavianov gamadrilov.

Intramuscular or aerogenic inoculation of baboons with Lassa virus reproduced a severe form of the disease which clinically, pathoanatomically and virologically resembles the severe form of human Lassa fever reported in the literature. The severity of monkey infection is demonstrated by rapid development of symptoms of general toxicity in the presence of fever, manifestations of hemorrhagic diathesis, high level of viremia and isolation of the causative agent from nasopharyngeal washings.

[The therapeutic efficacy of ribamidil and virazole in experimental Lassa fever in monkeys]. / Terapevticheskaia éffektivnost' ribamidila i virazola pri éksperimental'noi likhoradke Lassa u obez'ian.

The therapeutic efficacy of ribamydil and virasol was evaluated in experimental Lassa fever in monkeys which received these drugs at various intervals after the onset of the clinical illness. Ribamydil or virasol administered in the first day of fever protected from death 60% to 66% of the infected animals, but when the drugs were given 2 days after the onset of fever the number of survivors declined to 0.20%. When the treatment was started 4 days after the onset of fever none of the drugs prevented deaths of Lassa virus-infected baboons.

Detection of Lassa virus RNA in specimens from patients with Lassa fever by using the polymerase chain reaction.

Suitable oligonucleotide primers and probes were synthesized to amplify Lassa virus (Josiah strain)-specific nucleoprotein and glycoprotein gene fragments by using reverse transcription combined with the polymerase chain reaction (PCR). Our primers did not amplify the related lymphocytic choriomeningitis virus. By using PCR, about 50 50% tissue culture infective doses could be detected in the supernatant of infected cells. Furthermore, in all five serum specimens and four of five urine specimens of patients with acute Lassa fever, viral RNA could be demonstrated. Negative results were obtained with all serum and urine specimens of healthy subjects. Our data suggest that PCR may be applied as an alternative to virus isolation in the rapid diagnosis of Lassa fever.

[Lassa and Mozambique viruses: cross protection in experiments on mice and action of immunosuppressants on experimental infections]. / Virusy lassa i mozambik: perekrestnaia zashchita v opytakh na myshakh i deistvie immunosupressantov na éksperimental'nuiu infektsiiu.

The pathogenicity of Mozambique virus for random-bred mice and CBA mice was studied. In contrast to Lassa virus, intracerebral inoculation of newborn zandom-bred mice with Mozambique virus (1000 PFU/mouse) results in death of the animals. The pathogenic properties of both viruses for adult CBA mice were found to be similar: intracerebral inoculation of the viruses caused death of the animals within 6-8 days, but not intraperitoneal inoculation. With the latter, they produced a population of immunocompetent cells protecting syngeneic recipient mice against the lethal intracerebral inoculation of the homologous virus. Cross-protection experiments demonstrated that intraperitoneal inoculation of Lassa virus protected 70% of mice against the lethal infection with Mozambique virus, and intraperitoneal inoculation of Mozambique virus protected 45% of mice against Lassa virus. Cyclophosphamide exerted no protective effect in Mozambique virus-infected mice. Cyclosporin A exerted no therapeutic effect in mice lethally infected with Lassa or Mozambique virus.

[Effect of immunosuppression on the development and outcome of an acute infection in mice caused by administration of the Lassa virus]. / Vliianie immunosupressii na razvitie i iskhod ostroi infektsii, vyzyvaemoi u myshei vvedeniem virusa Lassa.

In experimental infection of mice with Lassa virus, the infectious virus could be detected in all the organs and brain tissues tested. Histopathological lesions were demonstrated in cerebral and spinal cord tissues only. Roentgen irradiation in a dose of 500 R and cyclophosphamide protected mice against a lethal Lassa virus dose. Cyclosporin A in various doses exerted no effect on the outcome of the acute infection. The adoptive transfer of splenocytes from mouse donors inoculated intraperitoneally prevented the development of lethal disease symptoms and death of mice-recipients. It is suggested that immunocompetent cells are involved in the development and outcome of experimental infection of mice with Lassa virus.

Identification of messenger RNA for nucleocapsid protein of Lassa virus in infected cells.

Single-stranded RNAs from Lassa virus-infected cells were fractionated by affinity chromatography on an oligo(dT)-cellulose column as well as by sucrose gradient centrifugation. Fractionated RNAs were translated in rabbit reticulocyte lysates, and translation products were precipitated with monoclonal antibodies to the nucleocapsid protein of Lassa virus. It has been shown that mRNA for the nucleocapsid protein is 15-16S and the RNA does not contain long if any poly(A) sequences.

Clinical virology of Lassa fever in hospitalized patients.

We measured levels of virus in sequential specimens from 137 patients with Lassa fever. The probability of fatal disease increased significantly with the level of viremia measured either on admission or during the course of illness. The odds ratio of death in patients with viremia greater than 10 TCID50/ml was 3.7 (90% confidence interval, 1.9-7.2). The same ratio in patients with viremia greater than 10 TCID50/ml and with levels of aspartate aminotransferase greater than or equal to 150 IU/liter was 21.5 (95% confidence interval, 5.2-99.0). Virus was found in throat cultures from 39% of viremic patients, compared with 14% of nonviremic patients (P less than .002); however, the level of virus was usually less than or equal to TCID50/ml. Fewer than 3% of patients were viruric during acute illness, and virus was isolated from three of three samples of cerebrospinal fluid. On admission, 53% of patients had IgG antibodies, and 67% had IgM antibodies. Recovery was not associated with the presence of either IgG or IgM. Virus was isolated from greater than 100 serum specimens that also contained high titers of IgG. Clinical Lassa fever was shown to be a disseminated systemic, primary viral infection, with an outcome highly associated with viremia but not with development of antibody.

Lassa virus hepatitis: a study of fatal Lassa fever in humans.

In order to explore the significance of a previous observation that the most important pathologic changes in fatal Lassa fever are hepatic, we have studied postmortem liver biopsies from 19 patients with fatal Lassa fever. We observed a vigorous macrophage response to cellular damage, but we found no evidence of lymphocyte infiltration in infected hepatic tissues. Using semi-quantitative estimates of liver cell damage, we found a wide range in the severity and progression of Lassa virus hepatitis in our fatal cases. We have classified for descriptive purposes three general nosopoeitic phases: active hepatocellular injury (less than 20% necrosis), continued damage and early recovery, and mitotic activity representing hepatic recovery. We conclude that the liver goes through cellular injury, necrosis and regeneration and any or all may be present at death. In no instance was the degree of hepatic damage sufficient to implicate hepatic failure, and all three phases were represented among our cases. We conclude that the hepatitis of Lassa fever in humans is not the primary cause of death.

[Pathogenicity of the Lassa virus for laboratory mice]. / Patogennost' virusa Lassa dlia laboratornykh myshei.

Pathogenicity for randombred and inbred mice of various age groups of the standard Lassa virus and the virus enriched with defective interfering particles (DIP) was studied. The standard Lassa virus inoculated intracerebrally caused 100% death of C3H/Sn mice aged up to 4 weeks and 60%-70% death of randombred white mice aged 3-4 weeks. BALB/C mice were found to be nonsusceptible to the virus, and its lethality for C57BL/6 and AKR mice varied within the range of 30%-60%. Lassa virus enriched with DIP caused no death of the susceptible animals and showed poor protective activity against the standard virus.

Kinetic study of platelets and fibrinogen in Lassa virus-infected monkeys and early pathologic events in Mopeia virus-infected monkeys.

The rhesus monkey, an established model of Lassa fever, was used to study hematologic and hemostatic aspects of Lassa fever and whether Mopeia (also known as Mozambique) virus induces any cellular damage in this model. Six days after subcutaneous injection of 10(3.48) plaque forming units (PFU) of Lassa virus (Josiah strain) one group of monkeys received an intravenous injection of 111In-labeled allogeneic platelets and another group received 125I-labeled alogeneic fibrinogen. Lassa virus-infected monkeys developed a severe clinical illness with high viremia and typical pathology. Lassa antigen was found in most tissues using a Lassa nucleocapsid-specific monoclonal antibody. Platelet counts remained within normal limits. Platelet and fibrinogen kinetics were similar in infected and control animals. Hematologic and hemostatic changes indicate that disseminated intravascular coagulation plays no role in this model of Lassa fever. Levels of plasma fibronectin were reduced in Lassa-infected monkeys. Mopeia virus-infected monkeys were normothemic, aviremic, and there was no detection of Mopeia antigen in any tissues using polyclonal or monoclonal antibodies. Mopeia virus was recovered from the spleen of one monkey. Mopeia virus was associated with hepatocellular and renal tubular damage.

Endemic Lassa fever in Liberia. III. Characterization of Lassa virus isolates.

Sixty-three virus isolates were obtained by inoculation of Vero cells with sera from 50 hospital in-patients in Liberia with acute febrile illnesses. 57 of the isolates were presumptively identified as Lassa virus (LV) by direct fluorescent antibody (DFA) staining of inoculated Vero cells. These, and six additional isolates obtained only by titration of supernatant fluids from inoculated Vero cells, were definitively identified as LV in a neutralization test. Two additional LV isolates were obtained from a patient's sera from Nigeria. By cross-neutralization tests, the Nigerian LV strains were serologically identical to the prototype Nigerian LV strain (PP) but were distinct from both a reference LV strain from Sierra Leone (SL), and from the Liberian (LIB) strains isolated in this study. The LIB and SL strains were closely related to each other, but not to the Nigerian LV strains. LIB LV strains were tested for virulence in strain 2 and 13 guinea-pigs, and a spectrum of virulence was observed which correlated only approximately with disease severity for human patients. Two human-lethal isolates killed all inoculated strain 2 and 13 guinea-pigs, whereas nine isolates from mildly ill patients were benign for guinea-pigs. Yet some LV isolates from severely ill or lethally infected patients, especially those from pregnant women and infants, were totally benign for guinea-pigs. These data suggest that antigenically distinct LV strains exist in nature, and that antigenically indistinguishable LV isolates may differ in virulence potential for various hosts.