Detection of IL-20R1 and IL-20R2 mRNA inC57BL/6 Mice Astroglial Cells and Brain Cortex Following LPS Stimulation

Astrocytes, which comprise 90% of overall brain mass, are involved in brain immunity. These cells represent the non-professional class of CNS-resident APCs and may promote or inhibit CNS inflammation depending on the cytokines they secrete. IL-10 family of cytokines and their receptors, IL-20R1 and IL-20R2, may have a role in shifting astrocytes to a neuroprotective or neurodegenerative function. To address the expression of IL-20R1 and IL-20R2 cytokine receptors in astrocytes and brain cortex of C57BL/6 mice. We investigated the expression of IL-20R1 and IL-20R2 in C57BL/6 mice astroglial cells and brain cortex in response to lipopolysaccharide [LPS], using reverse-transcription polymerase chain reaction [RTPCR] method. Astrocytes were able to express IL-20R1 and IL-20R2 mRNA not only in response to LPS stimulation but also in the absence of LPS. Furthermore, we found the expression of IL-20R1 and IL-20R2 mRNA in the cortex of adult C57BL/6 mice. IL-20R1 and IL-20R2 are constitutively express in the brain. Since most neuropathological processes involve astrocytes and inflammatory cytokines, these findings have important implications for future therapeutic strategies

Study of the expressions of IL-20R1 and IL-20R2 in C57BL/6 mice astroglial cells

Astrocytes are the most abundant glial cell type. They may promote or inhibit CNS inflammation depending on which cytokines are secreted. Astrocytes also have immune roles. IL-19, IL-20, and IL-24 activate a heterodimer receptor composed of the IL-20R1 alpha-chain and the IL-20R2 beta-chain. It has long been considered that signaling by these receptor complexes affects immunological reactions, however the biological functions of IL-20R1 and IL-20R2 in the brain remain unclear. As the first step to address the role of these cytokine receptors in the brain, in this study we have researched the expressions of IL-20R1 and IL-20R2 in C57BL/6 mice astrocytes. We examined expressions of IL-20R1 and IL-20R2 proteins in mice astroglial cells and in the 1321N1 astrocytoma cell line in response to MOG, LPS and GM-CSF by flow cytometry. The effect of LPS on mRNA expression of IL-20R1 and IL-20R2 was investigated by RT-PCR. We provide, for the first time, evidence that astrocytes expressed IL-20R1 and IL-20R2 mRNA not only in response to LPS stimulation but also in unstimulated astrocytes. We did not observe the expressions of IL-20R1 and IL-20R2 proteins in mice astroglial cells and the 1321N1 astrocytoma cell line. IL-20R1 and IL-20R2 mRNA are constitutively expressed in astrocytes. Because the majority of neuropathological processes involve astrocytes and inflammatory cytokines, the results of this study, which are reported for the first time, have important implications for future research

NOS2A gene polymorphism and its association with nitric oxide production in peripheral blood leukocytes of normal individuals

In peripheral blood polymorphnuclear and mononuclear cells Nitric Oxide [NO] could be synthesized by an enzyme called inducible NO synthase [iNOS]. iNOS gene [NOS2A] is located on chromosome 17 at position 17q11.2-q12. NO is released during inflammatory responses. In the present studies the frequency of NOS2A gene polymorphisms and their effects on NO production were investigated in the Peripheral Blood polymorphonuclear and mononuclear cells of normal individuals. In this study the frequency of NOS2A gene polymorphisms at positions -1659 C/T and +150 C/T of 232 normal subjects were investigated using PCR-Allele specific and PCR-RFLP methods, respectively. To study the effect of 1659C/T and +150C/T polymorphisms on NO production, polymorphonuclear and mononuclear cells of peripheral blood of 92 normal subjects were isolated and then stimulated by E.coli culture supernatants [ATCC 25922] for induction of iNOS enzyme and NO production. After 24h, the level of NO production in the culture supernatant were measured by Griess reaction. Polymorphisms as mentioned above were also studied in these normal cases. The results showed no significant difference in the level of NO production of various genotypes in the polymorphonuclear and mononuclear cells of peripheral blood of normal subjects. Our results indicated no significant correlation between NOS2A genotypes and NO production. No significant difference was observed between Gambia and China normal population and normal subjects of this study in NOS2A- 1659 C/T and +150 C/T polymorphisms. In Iran these differences are due to the genetic and ethnic differences among the studied populations, which indicates the importance of NOS2A polymorphism in the NO production, suggesting further studies in other ethnic groups