To understand the potential mechanisms of dietary protein on intestinal and host health, we studied the immunomodulatory effects of isocaloric diets with high or low crude protein (CP) contents on young adult Sprague Dawley (SD) rats. A total of 180 healthy male rats were randomly assigned to six groups (six replicate pens per treatment with five rats per pen) and fed diets with 10% CP, 14% CP, 20% CP (control), 28% CP, 38% CP, and 50% CP. Compared with the control diet, the rats fed the 14% CP diet significantly elevated lymphocyte cell counts in the peripheral blood and ileum, whereas the 38% CP diet significantly activated the expression of the TLR4/NF-κB signaling pathway in the colonic mucosa (p + T, B, and NK cells in the peripheral blood and the colonic mucosal expression of IL-8, TNF-α, and TGF-β. Overall, rats fed the 14% CP diet enhanced host immunity by increasing the numbers of immune cells, and the immunological state and growth of SD rats were negatively impacted by the diet containing 50% CP.