To explore the immune function of C-type lectin in shrimp, one recombinant C-type lectin (LvLec) was injected into Litopenaeus vannamei. There were four treatments in the experiment: saline group (as control group), recombinant C-type lectin group (LvLec, 1 mg mL−1), Vibrio harveyi group (V. harveyi, 106 cfu mL−1) and recombinant C-type lectin combined with Vibrio harveyi group (LvLec + V. harveyi, 1 mg mL−1 + 106 cfu mL−1). The sampling time was set at 0, 3, 6, 9, 12, 24 h after the injection. The results showed that the total hemocyte count decreased significantly and the phagocytic activity improved notably after the injection of LvLec, V. harveyi or LvLec + V. harveyi. Prophenoloxidase (proPO) activity decreased, while phenoloxidase (PO) activity increased and the changing degree of each group exhibited a significant difference. The hemagglutinating activity and bacteriolytic activity improved significantly, while the antimicrobial activity did not show a remarkable change in all of the groups. There were also changes that occurred in the levels of second messengers (cAMP, cGMP) and protein kinase (PKA, PKG). After the injection of LvLec, V. harveyi or LvLec + V. harveyi, the concentration of cGMP and PKA increased significantly, while the concentration of cAMP and PKG did not change remarkably. The results above suggested that rLvLec could induce nonspecific immune response, including phagocytosis, release of PO, hemagglutination and bacteriolysis through cGMP-PKA pathway in vivo.