Detection of substance P immunoreactivity in human peripheral leukocytes

The neuropeptide substance P (SP) has a marked proinflammatory effect and modulates the immune response. In this study, we tested the hypothesis that human peripheral leukocytes contain SP. Resting peripheral leukocytes collected from healthy volunteers (n = 20) were studied by applying a SP rabbit polyclonal antiserum to both flow cytometry and immunohistochemistry. To identify possible changes in Sp expression, we also analyzed both activated T-lymphocytes (cell cultures; n = 5 normal subjects) and neoplastic hematologic samples of different types of leukemias. Flow cytometry showed that normal granulocytes and monocytes contained SP, whereas lymphocytes were generally negative (or weakly positive) with the exception of a few (10-20%) positive subsets. In comparison, activated T-lymphocytes were markedly immunolabeled by SP as well as samples from neoplastic patients demonstrated strong SP immunoreactivity in all cell lineages. This pattern was confirmed by immunohistochemistry on cytospins. Our results support a potential role for SP-mediated immunomodulatory mechanisms both in normal and pathological conditions.