We have proposed that inappropriate induction of progrmmed cell death (PCD) or apoptosis, a physiological cell-suicide process, may play a role in the pathonesis of AIDS. This model has been supported by several reports of abnormal levels of PCD in vitro in both CD4+ and CD8+ T cells from human immuno iency virus type 1 (HlV-1)-infected persons. To further assess the nce of such a process In

Despite recent technical improvements, the construction of recombinant adenovirus vectors remains a time-consuming procedure which requires extensive manipulations of the viral genome in both Escherichia coli and eukaryotic cells. This report describes a novel system based on the cloning and manipulation of the full-length adenovirus genome as a stable plasmid in E. coli, by using the bacterial homologous recombination machinery. The efficiency and flexibility of the method are illustrated by the cloning of the wild-type adenovirus type 5 genome, the insertion of a constitutive promoter upstream from the E3 region, the replacement of the E1 region by an exogenous expression cassette, and the deletion of the E1 region. All recombinant viral DNAs were shown to be fully infectious in permissive cells, and the modified E3 region or the inserted foreign gene was correctly expressed in the infected cells.

E1-deleted adenovirus (Ad) vectors expressing the human to the induction of a cellular immune response to the lacZ coagulation factor IX (hFIX) or the bacterial ␤-galactoantigen. In contrast, absence of detectable levels of serum sidase (lacZ) were injected intravenously into various hFIX in immunocompetent animals was not associated with strains of immunocompetent (C57Bl/6, BALB/c, CD1, a loss of viral DNA but was strictly correlated with the CBA/J, C3H) and immunodeficient (BALB/c-nu/nu, induction of anti-hFIX antibodies. Surprisingly, anti-hFIX C57Bl/6-nu/nu, SCID, NIH-bg-nu-xid) mice. Regular analyantibodies were never detected in C57Bl/6 mice, leading sis of mouse sera and tissues showed a persistent to prolonged detection of hFIX. These results suggest that expression of both transgenes in immunodeficient mice, cellular immunity to viral antigens plays a minor role in the while detection diminished very rapidly in immunocompetearly extinction of transgene expression and illustrate the ent mice. The mechanisms responsible for the transient influence of the cellular (eg lacZ) or humoral (eg hFIX) detection of the two transgenes were however not identimmunity to transgene-encoded products on the persistical. Rapid decline of lacZ expression was correlated with a ence of transgene expression. rapid decrease of viral DNA sequences, and consequently

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