Different routes of vaccination with rAd induce protective anti-tumor immunity. B6 mice were immunized intraperitoneally (i.p.) (n = 8), intramuscularly (i.m.) (n = 8), or subcutaneously (s.c.) (n = 8) with rAd-1, immunized with PBS/BSA (n = 8), or immunized i.p. with rAd-2 (n = 8). Two weeks later, the mice received a subcutaneous challenge of 0.4 × 106 Ad5E1A + Ras cells. Mice immunized intraperitoneally, intramuscularly, or subcutaneously with rAd-1 are protected against Ad5E1A + EJras cells.