The cross-talk between the endocrine and the immune systems mediated by a wide array of hormones, cytokines, and neuromodulators is heightened during disease, stress, and presumably, during pregnancy. Adrenocorticotropin (ACTH) and nitric oxide (NO) are two immunomodulators that are also produced from lymphocytes and contribute to the immunomodulation. Thus, we investigated whether the heightened bidirectional communication between the immune and the endocrine systems observed during pregnancy is reflected in production of ACTH and NO from peripheral bovine lymphocytes and if any temporal correlation exists between them. Adrenocorticotropin was analyzed using a sandwich immunoradiometric assay, and nitrite and nitrate (a measure of NO) were estimated in supernatants of cultured peripheral blood lymphocytes (PBLs) using a colorimetric assay based on the Griess reaction. A significantly high secretion of ACTH and NO was noticed from PBLs in all stages of pregnancy compared to that in cyclic and cystic cows. Increased secretory capacity was noticed as early as 7 days after conception, which reached as much as 600% that of nonpregnant animals between Days 90–120 of gestation. Adrenocorticotropin and NO decline 1 mo before the expected time of parturition. Unlike those from cyclic animals, PBLs from pregnant cows were refractory to stimulation by PHA-M (Phytohemagglutinin) and corticotropin-releasing hormone. A strong correlation was observed between ACTH and NO secretion from PBLs in pregnant, in cyclic, and in cystic cows. To our knowledge, this is the first evidence elucidating the induction of ACTH and NO from PBLs during pregnancy, and it implies a new role for ACTH and NO secreted from PBLs in recognition and, probably, maintenance of pregnancy.