The C3 subtypes are differentially regulated after immunostimulation in rainbow trout, but head kidney macrophages do not contribute to C3 transcription

The complement system plays key roles in innate and adaptive immunity through mediating phagocytosis, chemotaxis and cell lysis. Complement component C3 is a central component in the complement cascade and belongs to the acute-phase proteins whose synthesis increase immediately upon inflammatory stimuli. The liver is the main producer of C3 and it is a well-known fact that the mammalian monocyte-macrophage lineage is a major contributor to extrahepatic C3. Immunomodulators, such as LPS and β-glucan, can stimulate complement, lysozyme, natural killer cells and antibody responses in fish, thus enhancing the resistance to bacterial pathogens and parasitic infections. The aim of this study was to assess the effects of LPS and β-glucan on the expression of interleukins (IL-1β1, IL-1β2 and IL-6) and the modulated expression of C3 subtypes (C3-1, C3-3 and C3-4) in the rainbow trout (Oncorhynchus mykiss) using real-time RT-PCR. From in vitro studies, we demonstrated that head kidney macrophages from rainbow trout and Atlantic salmon showed no basal transcription of C3. After immunostimulation, the cells responded by increased levels of ILs, but transcription of C3 was not induced. In contrast to the in vitro findings, the rainbow trout complement C3 subtypes were differentially regulated 48 h after in vivo stimulation with LPS and β-glucan. These results support the previous findings of absence of C3 in macrophages of the spotted wolffish (Anarhichas minor) and is the first functional study showing differential regulation of the C3 subtypes in any vertebrate.