The aim of this study was to investigate the effects of deficient or excess of dietary threonine (Thr) levels on intestinal integrity and barrier function of broilers. A total of 432 1-day-old commercial broilers (Arbor Acre) were assigned to four experiment groups consisting of six replicates of 18 birds. The treatments were designed as follows: 85%, 100%, 125% and 150% of NRC (Nutrient requirements of poultry (9th edn). Washington, DC: The National Academies Press, 1994) recommendations. The results indicated that expressions of jejunal and ileal secretory immunoglobulin A (sIgA) mRNA were increased linearly or quadratically by increasing Thr (p<.05), and the highest sIgA mRNA abundance was obtained in 125% Thr level. Likewise, the intestinal sIgA content showed similar increasing trend with the intestinal sIgA gene expression in this instance. The high level of Thr inclusion upregulated mucin 2 (MUC2) mRNA expression in the jejunum and ileum (p<.05). In addition, on day 21, the expression levels of jejunal zonula occludens-2 (ZO-2) and ileal zonula occludens-1 (ZO-1) decreased then increased with increasing Thr level (p<.05), whereas, the mRNA expressions of occludin in the jejunum and ileum had no significant difference amongst groups (p>.05). On day 42, Thr treatments did not affect the mRNA abundance of measured genes in the jejunum and ileum (p>.05). These findings suggested that Thr might be a nutrient immunomodulator that affects intestinal barrier function, moreover, 125% of the NRC (1994) recommendations Thr level was optimum.