Regulatory peptide immunoreactivities reported in the upper respiratory system of man include vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM), which are co-localized in a network of fine varicose nerve fibers. The present study was undertaken to examine the possible occurrence and distribution of the recently described VIP-like peptide helospectin. Double immunofluorescence labelling showed that helospectin is co-localized with VIP and PHM. In addition to nerve fibers containing all three peptides, scattered nerve fibers were detected that were only immunoreactive to helospectin but not to VIP and/or PHM. The distribution of helospectin/VIP/PHM immunoreactive nerve fibers around blood vessels and in close relationship to seromucous glands indicates their possible involvement in the regulation of blood flow and secretion.