Recombinant vaccinia virus coexpressing the F protein of respiratory syncytial virus (RSV) and interleukin-4 (IL-4) does not inhibit the development of RSV-specific memory cytotoxic T lymphocytes, whereas priming is diminished in the presence of high levels of IL-2 or gamma interferon

Abstract

In order to investigate if immune responses to the fusion (F) protein of respiratory syncytial virus (RSV) could be influenced by cytokines, recombinant vaccinia viruses (rVV) carrying both the F gene of RSV and the gene for murine interleukin-2 (IL-2), IL-4, or gamma interferon (IFN-gamma) were constructed. In vitro characterization of rVV revealed that insertion of the cytokine gene into the VP37 locus of the vaccinia virus genome resulted in 100- to 1,000-fold higher expression than insertion of the same gene into the thymidine kinase (TK) locus. In comparison, only a two- to fivefold difference in the level of expression of the F protein was observed when the gene was inserted into either of these two loci. Mice vaccinated with rVV expressing the F protein and high levels of IL-2 or IFN-gamma cleared rVV more rapidly than mice inoculated with a control rVV and developed only low levels of RSV-specific serum antibody. In addition, these recombinants were much less effective at priming RSV-specific memory cytotoxic T lymphocytes (CTL) and IFN-gamma production by spleen cells than rVV expressing the F protein alone. In contrast, mice vaccinated with rVV expressing high levels of IL-4 showed signs of delayed rVV clearance. RSV-specific serum antibody responses were biased in favor of immunoglobulin G1 (IgG1) in these mice, as there was a significant reduction in IgG2a antibody responses compared with serum antibody responses in mice vaccinated with rVV expressing the F protein alone. However, vaccination with rVV expressing the F protein together with high levels of IL-4 did not alter the development of RSV-specific memory CTL or IFN-gamma production by RSV-restimulated splenocytes.

FIG. 1

Effect of cytokines on isotype-specific serum antibody responses to RSV. Mice were vaccinated with (A) VRBF (□), VV-F.mIL-2low (▪), VVF.mIFN-γlow (), or VV-F.mIL-4low () or with (B) VSCF (□), VV-F.mIL-2high (▪), VV-F.mIFN-γhigh (), or VV-F.mIL-4high (). ND, Not determined. Data are given as log₁₀ mean antibody titers ± SD.

FIG. 2

Effect of cytokines on priming of RSV-specific CTL. Shown is the cytotoxic activity of splenocytes from mice immunized with VSCF, VV-F.mIL-2high, VV-F.mIL-4high, VV-F.mIFN-γhigh, or VA-βgal (VAβ-gal) at different effector cell/target cell (E:T) ratios on BCH4 cells (H-2^d) (▪), RSV-infected L-929 cells (H-2^k) (□), or uninfected BALB/c fibroblasts (⧫).