Skewing of the Th1/Th2 responses in mice due to variation in the level of expression of an antigen in a recombinant BCG system

Abstract

In spite of rapid developments in the study of mycobacteria during the last two decades, tuberculosis (TB) has maintained its status as the leading killer among all infectious diseases. Extensive evidence exists to support a central role for a T-helper type 1 (Th1) immune response for protection against TB in mice and humans. Bacille Calmette-Guerin (BCG), the only vaccine against TB, although not perfect in its ability to protect against the adult form of TB, is a strong inducer of Th1 responses and is being increasingly used as a delivery vehicle for the presentation of foreign antigens to the immune system. It has been proposed that expression of immunodominant antigens or cytokine genes in BCG can enhance the ability of BCG to induce a Th1 immune response. Since dose of the antigen is considered as one of the parameters that influence the Th cell responses, the level of expression of the candidate antigen should influence the final Th response against the recombinant BCG (rBCG). In the present study, the effect of over-expression of a candidate antigen Antigen 85B (Ag 85B) in a rBCG system, on the Th-priming ability of BCG has been investigated in the murine model. BALB/c mice were immunized with three different rBCG constructs expressing Ag 85B to various levels. Induction of Th1/Th2 responses was analyzed by measuring levels of interferon-γ (Th1) and interleukin-10 (Th2) in antigen-stimulated splenocyte cultures and by quantifying the antigen-specific IgG2a (Th1) and IgG1 (Th2) antibody responses. By varying the level of expression of Ag 85B, specific immune responses against Ag 85B were observed to range from mixed Th1/Th2 to Th1. However, the BCG-specific immune responses in case of all rBCG-immunized animals remained predominantly Th1.