Metarhizium rileyi, a well-known entomopathogenic fungus, could open up new vistas in biological control of insect pests; however, due to its intrinsic shortcomings, such as long pathogenic process, its application is largely limited. To explore which process, the invasion or the following in vivo development, is the main factor responsible for the long pathogenic process, the lethal effect of M. rileyi against Spodoptera litura (Fabricius) was determined by conidial topical application and hyphae body injection, and the host immune response was also monitored. Results showed when larvae were inoculated by conidial topical application, the pathogenicity of M. rileyi varied greatly depending on the larval instar and conidia concentration, and LC₅₀ values ranged from 6.24 × 10⁶ to 6.06 × 10⁹ conidia/ml while LT₅₀ values fluctuated from 4.35 to 9.43 d. However, in vivo study showed when hyphal bodies (Hbs) of M. rileyi were injected into host hemocoel, they would not be recognized by the host's immune system as invaders. There were no significant differences in the hemocytes and phenoloxidase activity between the infected and control larvae at the initial 44 h, indicated that the fungus was able to successfully avoid the attack from the cellular and humoral immune systems, therefore, it could multiply freely in the hemocoel. The in vivo development time of M. rileyi tended to remain constant for 2–3 d regardless of the initial inoculated numbers. Considering no detectable defense response was observed during in vivo development, it can be concluded that host nonself-recognition system does not respond to the hemolymph borne-Hbs.