This experiment was conducted to culture broiler intestinal lymphocytes by the technology of in vitro cell culture, establish in vitro stress model of lipopolysaccharides (LPS) and H2O2, study the modulation of different levels of glutamine (Gln) on proliferative activity and antioxidation of broiler lymphocytes, and finally provide a theoretical basis for the improvement of intestinal health by Gln. Separate lymphocytes from broiler jejunum and make them into cell suspension. Culture the cells for 24 h in 200 μL solutions with 0, 10, 50, 100, 200 μg/mL Gln, respectively. Add LPS into the solutions and test cell proliferation by MTT method. Imitate the condition of oxidative stress by stimulating intestinal mucous membrane with 0.3 mmol/L H2O2. Collect the supernatant and test the oxidative and immune stress parameters. The results showed as follows: 1) the 100 μg/mL level of Gln had the most obvious inhibiting effect on lymphocyte proliferative activity. 2) The 50 and 100 μg/mL levels of Gln could significantly increase the activities of catalase (CAT) and superoxide dismutase (SOD) in intestinal lymphocytes of broilers (P<0.05), while the activities of CAT and SOD began to decrease when the Gln level was 200 μg/mL; the malondialdehyde (MDA) content in 0 μg/mL Gln group was significantly higher than that in the other groups (P<0.05), and there were no significant differences among Gln groups (P>0.05). 3) The immunoglobulin A (IgA) content in intestinal lymphocytes of broilers in 100 μg/mL Gln group was significantly higher than that in the other groups (P<0.05). In conclusion, the 100 μg/mL level of Gln has the most obvious effect on suppressing proliferative activity of intestinal lymphocytes of broilers, and it increases IgA content, which is good for the maintenance of immune system balance; 50 and 100 μg/mL level of Gln can significantly increase the activities of CAT and SOD, which is good for the maintenance of antioxidant function of broiler intestine.［Chinese Journal of Animal Nutrition，2011，23（2）：274-279］