| Cytokines are important immune-active molecules, and their roles in CNS have been paid much attention. Studies have shown that cytokines may be important humeral messengers that mediate immune information to CNS. It is important for neuroimmunomodulation to studying the sites and cells in the brain that inflammatory cytokines acts on. It is known that cytokines exert their biological functions through their high-affinity receptors that exist on the surface of cell. Thus, studies on change of inflammatory cytokines receptors expressions in the brains of immune model animals have important significance for revealing the nuclei and cells participating neuroimmunomodulation. The thesis first reviewed the recent progress in the study of three kinds of inflammatory cytokines (IL-1, IL-6 and TNF-a) and their receptors. And then in two parts including four experiments, we made a series of studies about inflammatory cytokine receptors.In the first part, we observed the changes of expressions of type I receptor of IL-1 in the rat and mouse brain after intraperitoneally administration of different kinds and doses antigens respectively. In the second part including two experiments we cloned rat IL-6R'S genes by PCR, expressed them in E.coli DH5 a and cos-7 cell, and produced IL-6R'S polyclonal antibody which is proved having more high liter, affinity and specificity1. The changes of expression of IL-1 receptor type I in the rat brain were observed by immunohistochemistry after LPS or SEB was administeredintraperitoneally. The results showed that the expressions of IL-1 receptor type6 ％ttI in normal adult rat brain were very widely, and there were intense immuno-positive cells of IL-1 receptor type I in septal area, medial preoptic area, neocortex, hippocampus, paraventricular nucleus of the hypothalamus, supraoptic nucleus, hypothalamic ventromedial nucleus, arcuate nucleus and median eminence. Comparing with controls, the amount and dyeing of immunopositive cells of IL-1 receptor type I in rat paraventricular nucleus of the hypothalamus and the supraoptic nucleus were significantly enhanced by intraperitoneally LPS or SEB injection (PO.05), and the dyeing areas and processes of positive cells were enlarged. The present results indicated that the paraventricular nucleus of the hypothalamus and the supraoptic nucleus might have important roles in neuroimmunomodulation.2.Following LPS or SEB was administered intraperitoneally, the expression of PCNA of splenic cells and IL-1 receptor type I in PVN and SON were observed by using immunocytochemistry in the mice. Double fluorescent labeling technique was used to determine the relationship of IL-1 receptor typeI co-expressions with arginine vasopressin or oxytocin. The results showed the expression of PCNA of mouse splenic cells was significantly enhanced by LPS or SEB compared with the controls (.PO.05 or 0.01) . The expressions of IL-1 receptor type I in mouse brain were widely distributed. And the expressions in mouse PVN and SON were significantly enhanced by an intraperitoneal administration of LPS or SEB (P<0.05 or 0.01) .The arginine vasopressin- and oxytocin-containing neurons in the PVN and SON also displayed immunofluo-rescence labeling for IL-1 receptor type I . The results suggest that the hypothalamic PVN and SON might participate in neuroimmunomodulation, and partial arginin-vasopressin- and oxytocin-containing neurons in these twonuclei might have a pivotal role.3. IL-6R gene was amplified by PCR from the brain cDNA library of the normal adult rat. Expression vectors of whole IL-6R gene and its carboxyl terminal partial gene sequence were constructed by PCR and gene recombination technique, respectively. After sequencing the vectors were transformed into E.coli DH5 a and recombinant fusion protein was expressed via induction of IPTG. Fusion protein coded by carboxyl terminal gene sequence was purified through glutathione agarose column. Finally we obtained normal adult rat IL... |
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