Survival of all animals depends on effective protection against infection. In Drosophila, opportunistic infection kills larvae if they lack the Rel/NF-κB proteins Dorsal and Dif. We have used tissue-specific expression of Dif and Dorsal to reveal that these Rel proteins act in three different tissues to defend larvae from infection. Dif and Dorsal act in circulating blood cells, where they are required autonomously to promote blood-cell survival and phagocytosis of microorganisms. We show that a major transcriptional target of Dorsal and Dif in blood cells is Drosophila IAP1, a gene protecting these cells from death. We find that in addition to their autonomous role in blood-cell survival, Dif and Dorsal also act in the fat body to produce factors that promote blood-cell viability. These Rel proteins act in the epidermis to prevent infection by maintaining a barrier to microbial entry. Dorsal or Dif in any one of the three tissues is sufficient to defend the animal from opportunistic infection. Thus Drosophila has a multi-pronged system of defense and each branch of this network requires Rel proteins. Based on similarities between Drosophila and mammals, we propose that a Rel-dependent network is an ancient and robust framework of animal immune systems.