John J. Cox

Phase I Trial of a CD8+ T-Cell Peptide Epitope-Based Vaccine for Infectious Mononucleosis

ABSTRACT A single blind, randomized, placebo-controlled, single-center phase I clinical trial of a CD8+ T-cell peptide epitope vaccine against infectious mononucleosis was conducted with 14 HLA B*0801-positive, Epstein-Barr virus (EBV)-seronegative adults. The vaccine comprised the HLA B*0801-restricted peptide epitope FLRGRAYGL and tetanus toxoid formulated in a water-in-oil adjuvant, Montanide ISA 720. FLRGRAYGL-specific responses were detected in 8/9 peptide-vaccine recipients and 0/4 placebo vaccine recipients by gamma interferon enzyme-linked immunospot assay and/or limiting-dilution analysis. The same T-cell receptor Vβ CDR3 sequence that is found in FLRGRAYGL-specific T cells from most EBV-seropositive individuals could also be detected in the peripheral blood of vaccine recipients. The vaccine was well tolerated, with the main side effect being mild to moderate injection site reactions. After a 2- to 12-year follow-up, 1/2 placebo vaccinees who acquired EBV developed infectious mononucleosis, whereas 4/4 vaccinees who acquired EBV after completing peptide vaccination seroconverted asymptomatically. Single-epitope vaccination did not predispose individuals to disease, nor did it significantly influence development of a normal repertoire of EBV-specific CD8+ T-cell responses following seroconversion.

Peptide based cytotoxic T-cell vaccines; delivery of multiple epitopes, help, memory and problems

Synthetic CD8+ cytotoxic T-lymphocyte (CTL) peptide epitope based vaccines are being developed against a number of human diseases. Here we describe extensive preclinical testing of peptide epitope vaccines formulated with a protein as a source of CD4 help and Montanide ISA 720, an adjuvant currently in human clinical trials. Such water-in-oil formulations could effectively co-deliver several peptide epitopes and simultaneously induce multiple independent CTL responses. The efficiency of CTL induction by some peptides was, however, dependent on the aqueous buffer conditions, with poor performance correlating with non-covalent peptide oligomerisation. Any of a number of proteins currently used in human vaccines could supply CD4 help and no difference in CTL induction was obtained if the CD4 response was amnestic or a primary. Peptide immunisation was found to induce long term CTL memory and the recall of protective responses did not depend on an amnestic CD4 response. Slow pyroglutamic acid formation and rapid oxidation of methionine residues was observed in water-in-oil formulations, however, the latter had no effect on CTL induction. These data highlight the need to monitor for potential deleterious chemical events and interpeptide interactions, but illustrate that peptide based vaccination can effectively deliver multiple epitopes, in conjunction with any protein, and induce protective memory.

Demographic Characteristics of a Reintroduced Elk Population in Kentucky

We used radiotelemetry and field observations to study survival and reproduction of 718 reintroduced elk (Gervus elaphus) in eastern Kentucky, USA, from 1997 to 2001. Capture-related injuries accounted for 49% of the transit and post-release mortality. Annual survival was high across all age and sex classes, and ranged from 0.90 (adult females) to 0.97 (yearling females). Calving rates increased from 66% in 1998 to 92% in 2000. A high nutritional plane may explain the relatively high reproduction among females bred as yearlings, consecutive-year pregnancies, and twinning. Such high survival and reproductive rates are characteristic of colonizing ungulate populations in areas devoid of predators and competitors. Future research should focus on Kentucky-born calves to more accurately determine the effects of meningeal worm (Prelaphostrongylus tenuis) and other factors on recruitment, colonization, and population establishment.

Florida Panther Habitat Use: New Approach to an Old Problem

Abstract Although the Florida panther (Puma concolor coryi) has a history of study spanning more than 2 decades, some researchers have criticized previous habitat analyses of the panther for using daytime telemetry locations to infer 24-hour habitat use, selective use of radiotagged animals, comparison of animal locations to an inappropriate set of available resources, use of land cover maps for time periods that poorly matched some animal locations, use of locations instead of the individual animal as the sampling unit, and failure to account for telemetry error. To address all but the first of these concerns, we analyzed over 57,000 radiolocations of 100 Florida panthers and 8 introduced Texas cougars (Puma concolor stanlyana) collected from 1981 to 2003 using a Euclidean distance-based analysis (EDA), a technique that measures linear distances between telemetry locations and habitat types to determine nonrandom habitat use. We matched panther locations to 1 of 3 land cover maps reflecting cover conditions spanning 23 years to determine second- and third-order habitat selection. Panthers selected a mosaic of habitats when establishing a home range, and selected forests and avoided open wetlands within their home range. We recommend that managers give forests highest consideration when conserving this endangered species and suggest the further evaluation of the habitat value of natural openings that occur in close proximity to forested panther range.

Vaccination strategies for managing brucellosis in Yellowstone bison.

Concerns over migratory bison (Bison bison) at Yellowstone National Park transmitting brucellosis (Brucella abortus) to cattle herds on adjacent lands led to proposals for bison vaccination. We developed an individual-based model to evaluate how brucellosis infection might respond under alternate vaccination strategies, including: (1) vaccination of female calves and yearlings captured at the park boundary when bison move outside the primary conservation area; (2) combining boundary vaccination with the remote delivery of vaccine to female calves and yearlings distributed throughout the park; and (3) vaccinating all female bison (including adults) during boundary capture and throughout the park using remote delivery of vaccine. Simulations suggested Alternative 3 would be most effective, with brucellosis seroprevalence decreasing by 66% (from 0.47 to 0.16) over a 30-year period resulting from 29% of the population receiving protection through vaccination. Under this alternative, bison would receive multiple vaccinations that extend the duration of vaccine protection and defend against recurring infection in latently infected animals. The initial decrease in population seroprevalence will likely be slow due to high initial seroprevalence (40-60%), long-lived antibodies, and the culling of some vaccinated bison that were subsequently exposed to field strain Brucella and reacted positively on serologic tests. Vaccination is unlikely to eradicate B. abortus from Yellowstone bison, but could be an effective tool for reducing the level of infection. Our approach and findings have applicability world-wide for managers dealing with intractable wildlife diseases that cross wildlife-livestock and wildlife-human interfaces and affect public health or economic well-being.

Factors affecting reproduction and population growth in a restored elk Cervus elaphus nelsoni population

In 1997, the Kentucky Department of Fish and Wildlife Resources began a restoration program intended to translocate 200 elk Cervus elaphus nelsoni per year over a nine-year period. Initially, the age structure of males in this restored elk population was heavily skewed toward the yearling age class. We examined the reproductive performance of this elk herd for two years. During 1998, the male:adult female ratio was 35:65 (corresponding to 54:100). In 1999, the male:adult female ratio was 45:62 (corresponding to 73:100). The proportion of yearling males declined from 89% in 1998 to 31% in 1999. We used radio telemetry to locate males and females during the rut and to document calf production. Calving rates during 1998 and 1999 were 53 and 92%, respectively. Post-release movements of adult cows (N = 22) to areas devoid of males averaged 21 km and ranged within 7–57 km. Temporary Allee effects may have been responsible for annual differences in calving rates. The calving season was 67 days when breeding was dominated by yearlings and 37 days when breeding was dominated by adults. A male age structure heavily skewed toward yearlings does not appear to limit population growth. Calving rates could be improved by reducing initial post-release wanderings of adult females. Distribution of potential mates may be more important to population growth rather than balanced sex ratios and age structures.

Small Mammal Response to Vegetation and Spoil Conditions on a Reclaimed Surface Mine in Eastern Kentucky

Abstract Ecologically effective mine reclamation is characterized by the return of pre-mining floral and faunal communities. Excessive soil compaction typically results in delayed succession and low species diversity on reclaimed mine lands. We compared small mammal abundance and diversity among three levels of compaction in reforestation plots on an eastern Kentucky surface mine during 2004 and 2005. Compaction levels included 1) no compaction (loose-dumped), 2) light compaction (strike-off), and 3) high compaction (standard reclamation). Peromyscus leucopus (White-footed Mouse) made up 98% (295 of 300) of all individuals captured. In 2004, loose-dumped plots had more White-footed Mice (n = 108, mean = 36, SE = 0.58) than high-compaction plots (n = 62, mean = 20.6, SE = 3.10). Strike-off plots had more White-footed Mice (n = 59; mean = 19.6, SE = 0.66) than loose-dumped (n = 46, mean = 15.3, SE = 1.45) or high-compaction (n = 20, mean = 6.6, SE = 2.19) plots in 2005. Canopy cover and large rocks that created crevices appear to have been the factors that most influenced White-footed Mouse abundance on our study sites. Low small-mammal species diversity across all treatments was likely due to the presence of low quality habitat resulting from a poorly developed ground layer and soil compared to that found in undisturbed forest. Additionally, an insufficient amount of time since reclamation for small-mammal colonization from surrounding forests and a relatively large matrix of non-forested reclaimed mineland between our plots and potential source habitats may have also limited small-mammal diversity. To promote biodiversity and provide better wildlife habitat, we suggest that mine operators consider using reclamation methods that promote surface and vegetation heterogeneity and connectivity to source habitats.

Food Habits of Reintroduced Elk in Southeastern Kentucky

Abstract Based on microhistological examinations of feces, Cervus elaphus (elk) from a reintroduced herd on the Cumberland Plateau in southeastern Kentucky exhibited an annual diet of grasses (24%), forbs (27%), and browse (32%). Diets shifted seasonally, possibly in response to availability and palatability. Forbs dominated the summer diet (34%), whereas grasses, forbs, and woody browse accounted for approximately equal portions of the fall diet. Grasses (40%), and browse (46%) dominated the diet during winter and spring, respectively. Grasses were eaten less during spring (10%) than during any other month. Nutritional quality does not appear to be limiting in this growing population.

MENINGEAL WORM IN A REINTRODUCED ELK POPULATION IN KENTUCKY

Meningeal worm (Parelaphostrongylus tenuis) has been implicated in the failure of several elk (Cervus elaphus) restoration attempts in the eastern United States. However, limited post-release monitoring and a paucity of published literature prevents a clear understanding of this parasites role in past failures. During winters of 1997–2001, the Kentucky Department of Fish and Wildlife Resources translocated 1,044 elk from western states to eastern Kentucky (USA) in an effort to restore a free-ranging population. We monitored 521 radio-collared elk over 4 yr to determine the impact meningeal worm had on population establishment. Thirty (23%) of 129 non-capture related mortalities were attributed to meningeal worm. Twenty-two (73%) of these meningeal worm-caused mortalities were animals <3 yr old. If younger elk born in Kentucky suffer higher mortality rates than older translocated elk, the population growth observed during the initial years of restoration may be temporary. Additional research is necessary to determine the influence meningeal worm will have on elk population growth in Kentucky.

Space and habitat use of moose in southwestern Sweden

Increasingly, wildlife managers and land managers are challenged to maintain the viability of large mammal populations. Although the moose (Alces alces) is an ecologically, economically, and culturally important ungulate species found throughout most of Sweden, little is known about its ecology throughout the southern part of the country. We collected baseline ecological data on spatial and habitat use patterns in urbanized southwestern Sweden by fitting 22 adult moose (13 F, nine M) with global positioning system (GPS) radio collars. Home range size of cows did not differ among seasons (P > 0.10); however, bulls had larger home ranges during fall than all other seasons (P < 0.010). Mean home range size of males during fall and spring was larger than females during any season (P < 0.010). We used a Euclidean distance-based approach to analyze multi-scale habitat selection by moose. Moose of both sexes and during all seasons selected boreal forest and mires when establishing a home range (P < 0.10). Moose had seasonal differences in habitat selection within their home range (P 0.001), and generally selected clear-cuts and early successional forests, mature coniferous forests, and glades, but avoided agricultural areas and open water. Habitat and space use characteristics of moose in our urbanizing study area were similar to characteristics reported for moose in forest-dominated landscapes of Fennoscandia.