The objective of this study was to investigate the effects of yeast culture for white distiller's grains on performance, immune function and intestinal mucosal structure of laying hens. Three hundred and twenty-four healthy 27-week-old Hyline laying hens with the similar body weight and egg production rate were randomly divided into three groups containing 6 replicates of 18 birds each. Laying hens in 3 groups were fed a corn-soybean meal type diet supplemented with 0 (control), 1% and 2% yeast culture for white distiller's grains, respectively. The trial contained one-week adaptation period and eight-week formal trial period. The results showed as follows:1) egg producing rate did not differ significantly among experimental groups of each period of time (P>0.05), but the egg producing rate at 1 to 4 weeks, 5 to 8 weeks and 1 to 8 weeks in 1% yeast culture for white distiller's grains group was increased by 1.79%, 2.07% and 1.93% compared with control group, respectively. Average daily feed intake at 5 to 8 weeks and 1 to 8 weeks in 1% yeast culture for white distiller's grains group had a higher trend than that in the control group (P<0.10). 2) At the end of week 4, serum globulin content in 2% yeast culture for white distiller's grains group was increased by 19.0% (P>0.05) and 27.2% (P<0.05) compared with control group and 1% yeast culture for white distiller's grains group, respectively, and serum phosphorus content in 1% and 2% yeast culture for white distiller's grains groups was significantly higher than that in control group (P<0.05). 3) At the end of week 4, serum immunoglobulin (Ig) G and IgM contents in 2% yeast culture for white distiller's grains group were significantly increased compared with control group and 1% yeast culture for white distiller's grains group (P<0.05); at the end of week 8, serum IgA and IgM contents in 2% yeast culture for white distiller's grains group and serum lysozyme (LZM) activity in 1% yeast culture for white distiller's grains group were significantly increased compared with control group (P<0.05). 4) Compared with control group and 2% yeast culture for white distiller's grains group, supplementation of 1% yeast culture for white distiller's grains significantly increased the villus height and crypt depth (P<0.05), and had a trend to increase the villus height to villus crypt ratio (V/C) in jejunum of laying hens (P<0.10). The overall results indicate that appropriate dosage (1% to 2%) yeast culture for white distiller's grains in the diet of laying hens can enhance the immune function and improve the intestinal mucosal structure.