Kent D. Wagoner

Analysis of Human Peripheral Blood Samples from Fatal and Nonfatal Cases of Ebola (Sudan) Hemorrhagic Fever: Cellular Responses, Virus Load, and Nitric Oxide Levels

ABSTRACT Peripheral blood samples obtained from patients during an outbreak of Ebola virus (Sudan species) disease in Uganda in 2000 were used to phenotype peripheral blood mononuclear cells (PBMC), quantitate gene expression, measure antigenemia, and determine nitric oxide levels. It was determined that as the severity of disease increased in infected patients, there was a corresponding increase in antigenemia and leukopenia. Blood smears revealed thrombocytopenia, a left shift in neutrophils (in some cases degenerating), and atypical lymphocytes. Infected patients who died had reduced numbers of T cells, CD8+ T cells, and activated (HLA-DR+) CD8+ T cells, while the opposite was noted for patients who survived the disease. Expression levels of cytokines, Fas antigen, and Fas ligand (TaqMan quantitation) in PBMC from infected patients were not significantly different from those in uninfected patients (treated in the same isolation wards), nor was there a significant increase in expression compared to healthy volunteers (United States). This unresponsive state of PBMC from infected patients despite high levels of circulating antigen and virus replication suggests that some form of immunosuppression had developed. Ebola virus RNA levels (virus load) in PBMC specimens were found to be much higher in infected patients who died than patients who survived the disease. Similarly, blood levels of nitric oxide were much higher in fatal cases (increasing with disease severity), and extremely elevated levels (≥150 μM) would have negatively affected vascular tone and contributed to virus-induced shock.

Lassa Fever in Guinea: I. Epidemiology of Human Disease and Clinical Observations

The arenavirus Lassa is found in West Africa, where it sometimes causes a severe illness called Lassa fever. Lassa fever has been seldom investigated outside of a few hyperendemic regions, where the described epidemiology may differ from that in areas of low or moderate incidence of disease. Through a prospective cohort study, we investigated the epidemiology and clinical presentation of Lassa fever in Guinea, where the disease has been infrequently recognized. A surveillance system was established, and suspected cases were enrolled at five Guinean hospitals. Clinical observations were made, and blood was taken for enzyme-linked immunosorbent assay testing and isolation of Lassa virus. Lassa fever was confirmed in 22 (7%) of 311 suspected cases. Another 43 (14%) had Lassa IgG antibodies, indicating past exposure. Both sexes and a wide variety of age and ethnic groups were affected. The disease was more frequently found, and the IgG seroprevalence generally higher, in the southeastern forest region. In some areas, there were significant discrepancies between the incidence of Lassa fever and the prevalence of antibody. Clinical presentations between those with Lassa fever and other febrile illnesses were essentially indistinguishable. Clinical predictors of a poor outcome were noted, but again were not specific for Lassa fever. Case-fatality rates for those with Lassa fever and non-Lassa febrile illnesses were 18% and 15%, respectively. Seasonal fluctuation in the incidence of Lassa fever was noted, but occurred similarly with non-Lassa febrile illnesses. Our results, perhaps typical of the scenario throughout much of West Africa, indicate Lassa virus infection to be widespread in certain areas of Guinea, but difficult to distinguish clinically.

DEMOGRAPHIC FACTORS ASSOCIATED WITH PREVALENCE OF ANTIBODY TO SIN NOMBRE VIRUS IN DEER MICE IN THE WESTERN UNITED STATES

We used long-term data collected for up to 10 yr (1994–2004) at 23 trapping arrays (i.e., webs and grids) in Arizona, Colorado, Montana, and New Mexico to examine demographic factors known or suspected to be associated with risk of infection with Sin Nombre virus (SNV) in its natural host, the deer mouse (Peromyscus maniculatus). Gender, age (mass), wounds or scars, season, and local relative population densities were statistically associated with the period prevalence of antibody (used as a marker of infection) to SNV in host populations. Nevertheless, antibody prevalence and some of the risk factors associated with antibody prevalence, such as relative population density, gender bias, and prevalence of wounding, varied significantly among sites and even between nearby trapping arrays at a single site. This suggests that local microsite-specific differences play an important role in determining relative risk of infection by SNV in rodents and, consequently, in humans. Deer mouse relative population density varied among sites and was positively and statistically associated with infection prevalence, an association that researchers conducting shorter-term studies failed to demonstrate. Both wounding and antibody prevalence increased with mass class in both males and females; this increase was much more pronounced in males than in females and wounding was more frequent in adult males than in adult females. Prevalence of wounding was greatest among seropositive deer mice, regardless of mass class, but many deer mice without detectable wounds or scars eventually became infected. Many of these patterns, which will be useful in the development of predictive models of disease risk to humans, were only detected through the application of data collected over a long (10-yr) period and with abundant replication.

SIN NOMBRE VIRUS INFECTION OF DEER MICE IN MONTANA: CHARACTERISTICS OF NEWLY INFECTED MICE, INCIDENCE, AND TEMPORAL PATTERN OF INFECTION

Sin Nombre virus (SNV), hosted by the deer mouse (Peromyscus maniculatus), is the principal cause of hantavirus pulmonary syndrome (HPS) in North America. To improve our understanding of factors that contribute to the occurrence of HPS, we conducted an extensive field study of the characteristics of newly infected (as determined by recent acquisition of antibody) deer mice and the temporal pattern of antibody acquisition (seroconversion) from 1994 through 2004 in Montana, USA. We sampled 6,584 individual deer mice, of which 2,747 were captured over multiple trapping periods. Among these 2,747 deer mice, we detected 171 instances of seroconversion. There was no relationship between seroconversion and the acquisition of scars. However, recently infected Montana deer mice were more likely to be older, more likely to be males, and more likely to be in breeding condition. In addition, recently infected male deer mice gained less weight over the 1-mo period following seroconversion than did those that did not acquire antibody, suggesting that SNV infection may have negatively impacted the health of infected rodents. Incidence was highly variable among years, and timing of infections was primarily associated with the breeding season (generally early spring through late fall).

Epizootiology of Sin Nombre and El Moro Canyon hantaviruses, southeastern Colorado, 1995-2000.

Sin Nombre virus (SNV) is an etiologic agent of hantavirus pulmonary syndrome. To better understand the natural history of this virus we studied population dynamics and temporal pattern of infection of its rodent hosts in southeastern Colorado (USA) from 1995 to 2000. We present evidence for the presence of two hantaviruses, SNV in deer mice (Peromyscus maniculatus) and El Moro Canyon virus in western harvest mice (Reithrodontomys megalotis), at our study sites. Sin Nombre virus appeared only sporadically in deer mouse populations; overall prevalence of antibody to SNV was 2.6%. El Moro Canyon virus was enzootic: seroconversions occurred throughout the year; antibody prevalence (11.9% overall) showed a delayed–density-dependent pattern, peaking as relative abundance of mice was declining. Males of both host species were more frequently infected than were females. An apparently lower mean survivorship (persistence at the trapping site) for SNV antibody-positive deer mice could indicate a detrimental effect of SNV on its host, but might also be explained by the fact that antibody-positive mice were older when first captured.

POPULATION DYNAMICS OF A DIVERSE RODENT ASSEMBLAGE IN MIXED GRASS-SHRUB HABITAT, SOUTHEASTERN COLORADO, 1995-2000

We followed seasonal and year-to-year population dynamics for a diverse rodent assemblage in a short-grass prairie ecosystem in southeastern Colorado (USA) for 6 yr. We captured 2,798 individual rodents (range, one to 812 individuals per species) belonging to 19 species. The two most common species, deer mice (Peromyscus maniculatus) and western harvest mice (Reithrodontomys megalotis), generally had population peaks in winter and nadirs in summer; several other murid species demonstrated autumn peaks and spring nadirs; heteromyids were infrequently captured in winter, and populations generally peaked in summer or autumn. Interannual trends indicated an interactive effect between temperature and precipitation. Conditions associated with low rodent populations or population declines were high precipitation during cold periods (autumn and winter) and low precipitation during warm periods (spring and summer). Severity of adverse effects varied by species. Heteromyids, for example, were apparently not negatively affected by the hot, dry spring and summer of 2000. Cross-correlations for the temporal series of relative population abundances between species pairs (which are affected by both seasonal and interannual population dynamics) revealed positive associations among most murids and among most heteromyids, but there were negative associations between murids and heteromyids. These results have important implications for those attempting to model population dynamics of rodent populations for purposes of predicting disease risk.

PREDICTION OF PEROMYSCUS MANICULATUS (DEER MOUSE) POPULATION DYNAMICS IN MONTANA, USA, USING SATELLITE-DRIVEN VEGETATION PRODUCTIVITY AND WEATHER DATA

Deer mice (Peromyscus maniculatus) are the main reservoir host for Sin Nombre virus, the primary etiologic agent of hantavirus pulmonary syndrome in North America. Sequential changes in weather and plant productivity (trophic cascades) have been noted as likely catalysts of deer mouse population irruptions, and monitoring and modeling of these phenomena may allow for development of early-warning systems for disease risk. Relationships among weather variables, satellite-derived vegetation productivity, and deer mouse populations were examined for a grassland site east of the Continental Divide and a sage-steppe site west of the Continental Divide in Montana, USA. We acquired monthly deer mouse population data for mid-1994 through 2007 from long-term study sites maintained for monitoring changes in hantavirus reservoir populations, and we compared these with monthly bioclimatology data from the same period and gross primary productivity data from the Moderate Resolution Imaging Spectroradiometer sensor for 2000–06. We used the Random Forests statistical learning technique to fit a series of predictive models based on temperature, precipitation, and vegetation productivity variables. Although we attempted several iterations of models, including incorporating lag effects and classifying rodent density by seasonal thresholds, our results showed no ability to predict rodent populations using vegetation productivity or weather data. We concluded that trophic cascade connections to rodent population levels may be weaker than originally supposed, may be specific to only certain climatic regions, or may not be detectable using remotely sensed vegetation productivity measures, although weather patterns and vegetation dynamics were positively correlated.

ASSOCIATION BETWEEN MOVEMENT AND SIN NOMBRE VIRUS (BUNYAVIRIDAE: HANTAVIRUS) INFECTION IN NORTH AMERICAN DEERMICE (PEROMYSCUS MANICULATUS) IN COLORADO

Capture data from long-term, mark-recapture studies were used to evaluate movements of North American deermice (Peromyscus maniculatus) on mark-recapture webs in Colorado with respect to Sin Nombre virus (SNV) infection status, age, sex, and trapping site. Latitude and longitude coordinates for each capture during the approximately 12-yr study were used to produce an individual minimum convex polygon (MCP) area representing the movements (not home range) of an individual mouse over time. These MCP areas were compared by SNV infection status (as determined by the presence of antibody), age, and sex. Antibody-negative deermice had significantly larger mean MCP areas than did antibody-positive mice. No differences in MCP area were found between male and female mice (either positive or negative). The smaller MCP areas of antibody-positive mice correspond to decreased movement by SNV-infected deermice on the trapping webs. These findings may indicate that SNV has a negative effect on movement, perhaps by reducing the health of infected deermice.

SPATIAL CLUSTERING OF MURID RODENTS INFECTED WITH HANTAVIRUSES: IMPLICATIONS FROM META‐ANALYSES

We applied a rigorous, quantitative methodology to the analysis of local-scale spatial clustering of multiple murid mice (brush mice, Peromyscus boylii; deer mice, P. maniculatus; pinon mice, P. truei; western harvest mice, Reithrodontomys megalotis) infected or uninfected with hantaviruses. Rodents were sampled longitudinally from 1994 to 2001 on 23 trapping webs at 10 locations in the southwestern United States. This study provided an opportunity to apply meta-analysis techniques to an important ecological question. There were sufficient captures by species on 199 occasions (three consecutive nights) to compare general use of space using multi-response permutation procedures (MRPP). The MRPP results were then used in meta-analyses by species to determine if overall effects of spatial clustering of hantavirus-infected mice as well as categorical effects (elevation, season, site, and state) were present. Based on MRPP analyses, overall spatial clustering of hantavirus-infected mice was most pronounced for brush mice, followed by the deer mouse. Meta-analyses indicated significant overall effects of spatial clustering and varying categorical effects (elevation, season, site, state) of infected mice for each species compared. The overlapping space use by rodents might be an important factor affecting the local transmission of several hantaviruses.

Dynamics of Oliveros Virus Infection in Rodents in Central Argentina

Oliveros virus (OLV) is an arenavirus hosted by the sigmodontine rodent, Necromys benefactus, in central Argentina. We report a 3-year longitudinal field study of the dynamics of OLV infection in host populations from 15 localities in two provinces on the central Argentine pampa. There was an overall 3-year period immunofluorescent antibody prevalence of 25% in the host population, and infected hosts were found throughout the study area. Spill-over infection into common sympatric species was rare. Infection dynamics exhibited many of the patterns seen for other rodent-borne arenaviruses and hantaviruses, but had some unique characteristics. Host population density was highest in autumn and lowest in spring, while antibody prevalence was highest in spring and lowest in autumn. Virus transmission was horizontal: infection was strongly associated with age, reaching 45% prevalence in the oldest individuals, and prevalence of infection was equal among male and female hosts. Infection may have been associated with scars, which were also approximately equally distributed among male and female Necromys.