Two experiments were conducted to study the effects of lignocellulose supplementation on immune function in layer pullets at different stages of growth. Four-wk-old pullets (Experiment 1) were fed a control, diet (Diet C); Diet C plus 1% mixed soluble/insoluble fiber (Diet MF), or plus 1% insoluble fiber (Diet IF). At 7.5 wk-of-age, heterophil phagocytosis, and oxidative burst in Groups MF (328.5 beads/100 cells; 4,330.0 ΔRFU; relative fluorescent units) and IF (350.3; 5,264.4) were greater (P < 0.05) than Controls (303.4; 3,509.0). At 8 wk-of-age, Group MF and IF relative weights of bursa of Fabricius (0.57 g/100 g BW; 0.58 g /100 g BW), thymus glands (0.77; 0.78), and areas of Peyer's patches (PP) (2.7 cm2; 2.9 cm2) were higher (P < 0.05) than Controls (bursa, 0.50 g; thymus, 0.70 g; PP area, 1.8 cm2). In Experiment 2, 10-wk-old pullets were fed a control diet or diets containing 1.5% MF or IF for 8 wk. At 14 wk-of-age IF pullets had higher (P < 0.05) heterophil phagocytosis efficiency (447.9 beads/100 cells) than Controls (376.4) and MF and IF had greater (P < 0.05) oxidative burst (1,302.9 and 1,857.7 ΔRFU) than Controls (744.1). At 17 wk-of-age MF and IF had increased (P < 0.05) proliferation of T-lymphocytes (ConcanavalinA-stimulated) (100.4 and 103.1% of unstimulated cells) and B-lymphocytes (lipopolysaccharide-stimulated) (122.4 and 129.0) than Controls (ConA, 79.4; lipopolysaccharide, 106.6). At 18 wk-of-age, IF pullets were heavier (1,607.5 g, P < 0.05) than Controls (1,506.5 g), had heavier (P < 0.05) bursa of Fabricius (1.12 g) than MF and Control groups (0.98 g; 0.92 g) and cecal tonsils of MF (0.38 g) and IF (0.39 g) weighted more (P < 0.05) than Controls (0.33 g). Number of jejunal and ileal PP (10.0) in IF pullets was higher (P < 0.05) than Controls (7.1). These results indicate that both MF and IF can improve development of the immune system of young and grower pullets during periods of maturation and involution of lymphoid organs.