Anti-Pol Protein polyclonal antibody

SIV vaccination studies in monkeys have revealed that Env-directed antibodies are critical for protection against virus acquisition, whereas CD8(+) T-cell responses against the Gag and Pal proteins contribute to control of viremia postinfection. However, designing a vaccine that strongly activates both arms of the immune system, is challenging for a variety of reasons, one being interference of Gag and Env responses. Bockl et al. have studied how to optimize CD8 T-cell responses against Gag, Pol and Nef in the presence of Env co-vaccination. Although Env coadministration suppressed anti-Gag CD8(+) responses, this effect could be counteracted by adjusting the molar ratio of the vaccines and by spatial or temporal separation of the Gag and Env antigens. These results demonstrate that optimal induction of antiviral CD8(+) responses requires careful optimization of vaccine design, composition and administration.

A synthetic gene consisting of the reverse transcriptase (RT) and integrase (IN) domains of human immunodeficiency virus type 1 (HIV-1) pol was constructed using codons most frequently used in humans. The humanized pol gave dramatically improved levels of Rev-independent, in vitro protein production in mammalian cells and elicited much stronger cellular immunity in rodents than did virus-derived gene. Specifically, BALB/c mice were immunized with plasmids and/or recombinant vaccinia virus constructs expressing the synthetic gene. High frequencies of Pot-specific T lymphocytes were detected in these animals by the gamma interferon enzyme-linked immunospot assay against pools of short overlapping peptides. Characterization of the stimulatory peptides from these pools indicates that the optimized gene constructs are able to effectively activate both CD4(+) and CD8(+) T cells. Immunization of rhesus macaques with DNA vaccines expressing the humanized pol coupled to a human tissue plasminogen activator leader sequence led to pronounced in vitro cytotoxic T-lymphocyte killing activities and enhanced levels of circulating Pol-specific T cells, comparable to those observed in HIV-1-infected human subjects. Thus, optimizing the immunogenic properties of HIV-1 Pot at the level of the gene sequence validates it as an antigen and provides an important step toward the construction of a potent pol-based HIV-1 vaccine component.