Abstract. For a closed, oriented, odd dimensional manifold X, we define the rho invariant ρ(X, E, H) for the twisted odd signature operator valued in a flat hermitian vector bundle E, where H = i j+1 H 2j+1 is an odd-degree closed differential form on X and H 2j+1 is a real-valued differential form of degree 2j + 1. We show that ρ(X, E, H) is independent of the choice of metrics on X and E and of the representative H in the cohomology class [H]. We establish some basic functorial properties of the twisted rho invariant. We express the twisted eta invariant in terms of spectral flow and the usual eta invariant. In particular, we get a simple expression for it on closed oriented 3-dimensional manifolds with a degree three flux form. A core technique used is our analogue of the Atiyah-Patodi-Singer theorem, which we establish for the twisted signature operator on a compact, oriented manifold with boundary. The homotopy invariance of the rho invariant ρ(X, E, H) is more delicate to establish, and is settled under further hypotheses on the fundamental group of X.