After hepatitis B virus (HBV) infectionThe incubation period of a viral disease refers to the time between infection and the onset of symptoms, and has been believed to represent the phase of active viral replication before an effective host immune response. This period is generally long after infection with noncytopathic viruses such as hepatitis B virus (HBV), 1 in contrast to other viral diseases. [2][3][4] The development of symptoms, which may occur up to 6 months after HBV infection, 5 has been assumed to mark the onset of viral control by the immune system. 1 However, the observation of marked reductions in virus levels before liver injury in the chimpanzee model of HBV infection, 6 and of cytotoxic T lymphocyte (CTL)-mediated viral clearance in the absence of cytolysis, in a transgenic mouse model of HBV infection, 7 has challenged the evidence that HBV control is predominantly caused by CTL-mediated tissue injury. It is, therefore, possible that immunologic events important for the control of HBV may already be present during the long incubation phase of primary infection. However, patients are usually only diagnosed after the development of symptoms: thus, it has been difficult to study the relationship between viral dynamics and liver injury, and whether the incubation phase of HBV infection in humans represents a period of ongoing viral replication in the presence of a deficient immune response. 8 During a single-source outbreak of HBV infection, in which 30 patients were infected through a skin-piercing procedure known as autohemotherapy, 9 a number of patients were identified before the onset of clinical hepatitis. The recent development of immunologic techniques to directly quantify virusspecific lymphocytes, 10,11 enabling the dynamics of CD8 and CD4 responses during viral infections to be investigated, provided a unique opportunity to study the interaction among virus, clinical disease, and host immune responses from the incubation phase of acute HBV infection.
PATIENTS AND METHODS
Patients.Five patients (all women, mean age 54 years, range 37-71) were identified during the incubation phase of acute hepatitis B, and were longitudinally followed up through the course of the disease. Their infection was diagnosed on the basis of finding a positive serum hepatitis B surface antigen (HBsAg), HBV DNA by polymerase chain reaction, normal or minimally raised serum alanine transaminase (ALT) levels, a marker of hepatocyte damage, 12 and no symptoms of clinical acute hepatitis. Each patient was infected with the same variant of HBV, as assessed by viral genome sequencing and