The current study was conducted to evaluate the effect of different levels of clove bud and ?-tocopherol acetate on performance, egg quality, blood biochemistry parameters, ileal microflora, small intestinal histology, immunological responses and antioxidant indices of blood and egg in laying hens. In this study, a total of 180 Loehmann brown hen layers of 43 wk of age were randomly distributed among 9 experimental of treatments with 4 replicates of 10 birds each. Dietary treatments consisted of three levels of clove bud (0, 0.2 and 0.4%) and three levels of ?-tocopherol acetate (0, 20 and 200 U/kg of diet) that fed during a 80 d feeding trial including 10 days for adaptation period and 70 days as main recording period. Performance parameters including feed intake, feed conversion ratio, egg production percentage, egg weight and egg mass were calculated in three times of d 35, 70 and total period of experiment and egg quality of two periods of d 35 and 70 d of trial. To investigate of immunological response, sheep red blood cells were injected in two times with 10 d interval, hens were bled 6 and 12 d after the first and second injection, respectively. Moreover, 10 d before the first SRBC injection, Newcastle disease virus were injected and then blood sera were taken 6 d after injection. In addition, differential leukocyte counts were evaluated at the end of experiment. Furthermore, at the end of experiment, two birds of each cage were exsanguinated to evaluate the relative weight of liver, spleen as well as pancreas and also to sample ileal content and jejunal morphology. The results showed that inclusion of 0.2 % of clove bud led to a reduction in ileal Escherichia coli and Salmonella population (P 0.05), an improvement in intestinal health index and antibody titer against Newcastle disease virus (P 0.01), a reduction in malondialdehyde concentration of yolk, an increase in relative weight of spleen and pancreas as well as an improvement in feed conversion ratio (P=0.21). Dietary supplementation of 0.4% clove bud powder caused a decrease in cholesterol concentration of plasma and egg (P 0.05), triglyceride (P=0.06) and malondialdehyde of plasma (P 0.05), enzyme activity of SGOT and SGPT (P 0.01), heterophil to lymphocyte ratio (P 0.05), relative weight of liver (P 0.05), an improvement in antibody titer against SRBC (P 0.01) as well as egg quality (P 0.05). supplementation of the highest level of ?-tochopherol acetate led to a reduction in enzyme activity of SGPT and SGOT (P 0.01), a decrease in heterophil to lymphocyte ratio (P 0.05), a decrease in cholesterol (P 0.05) and malondialdehyde of egg yolk (P 0.01), an improvement in intestinal health index as well as relative weight of spleen and pancreas and antibody titer against SRBC (P 0.05). Additionally, an increase in dietary ?-tocopherol acetate significantly increased vitamin E concentration of plasma and egg (P 0.01). In general, results indicated that although inclusion of different levels of clove bud and ?-tocopherol acetate had no significant effects on performance parameters, supplementation of two antioxidant composition could improve intestinal health indices, immunological functions as well as antioxidant responses. Keywords: Laying hens, clove bud, vitamin E, performance, egg quality, immunological responses and lipid peroxidation