Expression of the murine interleukin-4 gene in an attenuated aroA strain of Salmonella typhimurium: persistence and immune response in BALB/c mice and susceptibility to macrophage killing

Abstract

Cytokines are potentially useful in vaccination as adjuvants or modulators of the type of response induced. The work below describes the expression of a cloned cytokine gene for murine interleukin-4 (mIL-4) by a live vaccine vector, an attenuated aroA strain (SL7207) of Salmonella typhimurium, in a murine model system. SL7207 was used as a carrier for two different high-level expression vectors. Both resulting strains, designated SL7207(pOmpAmIL-4) and SL7207(pKKmIL-4), expressed the cloned gene product as monitored by both immunological and biological assays. However, SL7207(pOmpAmIL-4) produced mIL-4 at higher levels and was more stable in vitro than SL7207(pKKmIL-4). When SL7207(pOmpAmIL-4) was used as a live vaccine in BALB/c mice, this strain grew and survived at higher levels than the parental attenuated strain or empty plasmid-carrying strain in spleens, livers, and intestines. This difference in growth and survival did not appear to be caused by alterations in specific lymphocyte-mediated anti-Salmonella immune responses such as delayed-type hypersensitivity or serum antibody as measured by enzyme-linked immunosorbent assay; such alterations have been induced by IL-4 administration in other in vivo systems, and the lack of effect here may reflect the fact that IL-4 is not secreted from the bacteria in large quantities, most of the cytokine being in the cytoplasmic-membrane-bound fraction. Conversely, the ability of mouse macrophages to kill the bacteria in vitro was inhibited by bacterial production of mIL-4. This reduction in macrophage killing activity suggests that bacterial production of mIL-4 may be detrimental to host defense against Salmonella infection and may explain the enhanced bacterial growth and survival in vivo.