John Spelbring

Experimental Studies on Subclinical Hepatitis E Virus Infection in Cynomolgus Macaques

Serial subclinical transmission among susceptible humans may serve as a reservoir of hepatitis E virus (HEV) in areas in which HEV is endemic. This hypothesis was investigated in an experimental primate model. Four groups of 4 cynomolgus macaques each were inoculated intravenously with 10(4)-10(5) (group 1), 10-100 (group 2), and 1-10 (group 3) cynomolgus macaque HEV infectious doses. All 4 animals in group 1 had clinical disease marked by alanine aminotransferase (ALT) elevation, fecal virus excretion, viremia, and seroconversion. Of the animals in groups 2 and 3, only 1 had evidence of biochemical hepatitis, although most had virus excretion and viremia (3 animals each in groups 2 and 3), and evidence of seroconversion (1 animal in group 2 and 3 animals in group 3). Viral genomic titers in stool specimens of animals with or without ALT elevation were similar. Infectivity studies confirmed the viability and transmission potential of the virus excreted by animals without ALT elevation. These data suggest that subclinical HEV infection may represent an HEV reservoir.

Protective Efficacy of Hepatitis E Virus DNA Vaccine Administered by Gene Gun in the Cynomolgus Macaque Model of Infection

The protective efficacy of a DNA vaccine against hepatitis E virus (HEV) infection was tested in cynomolgus macaques (cynos) vaccinated with a plasmid containing a full-length HEV open-reading frame 2 (ORF2) sequence (Burmese strain) and subsequently challenged with a heterologous strain of HEV (Mexican strain). Cynos administered vaccine by gene gun developed antibodies to HEV (anti-HEV), whereas cynos administered vaccine by intradermal injections and cynos administered a mock DNA construct did not develop anti-HEV. Anti-HEV-positive cynos were protected from HEV infection after challenge with an inoculum that produced infection in the anti-HEV-negative cynos. These results indicate that DNA vaccine with HEV ORF2 administered by gene gun is protective against a heterologous viral challenge.

Hepatitis E virus infection in chimpanzees: a retrospective analysis

Summary. Different patterns of disease were observed among 11 chimpanzees who were inoculated intravenously with hepatitis E virus (HEV) positive fecal specimens from four different outbreaks (Nepal 1981, Uzbekistan 1981, Pakistan 1985, and Mexico 1986). Five chimpanzees had marginal or no liver enzyme elevations within 70 days of inoculation. Two of the chimpanzees had limited viremia, but did not produce detectable antibody. The four remaining chimpanzees had liver enzyme elevations, viral shedding, viremia, seroconversion to anti-HEV, and detectable HEV antigen in liver biopsy specimens. These results may reflect the range of infection patterns that develop in humans after natural exposure to the HEV.

DNA vaccination against hepatitis E virus infection in cynomolgus macaques

Background : The feasibility of DNA vaccination against hepatitis E in non‐human primates has not been evaluated. In the present study a full‐length hepatitis E virus (HEV) open reading frame (ORF)2 (Burmese strain) was assembled, cloned, and used for genetic immunization of cynomolgus macaques (cynos), which were subsequently challenged with a heterologous HEV strain (Mexico).