ABSTRACT
In this study, capability of Vero cells for growth on FibraCel disks were compared on 3 kinds of microcarriers including Cytodex-1, Cutodex-3, and Sigma Solohill in 500 ml Spinner flasks in both serum contained medium [DMEM+10% Fetal Calf Serum, FCS] and serum-free medium [VP-SFM]. The propagation of fixed PV [Pasteur Virus] strain of rabies in Vero cells, for production of rabies vaccine, grown under the above conditions were studied and compared. Stepwise perfusion mode in growth phase and batch mode were applied in the virus production step by the use of M199 +0.2% Bovine Serum Albumin [BSA] and VP-SFM as serum containing and serum-free media, respectively. The available surface area provided by the carriers, and primary cell density in the experiments were assumed the same [about 12,000 cm[2] and 12,500 cells/cm[2], respectively]. The highest cell density was achieved on FibraCel disks in DMEM equal to 7.1 +/- 0.7_10[6] cells /ml on day 9, while the lowest cell density was obtianed on Cytodex-3 in VP-SFM equal to 2.91 +/- 0.2_10[6] cells/ml. The highest virus titer [55.18 +/- 4.4_10[6] Fluorescent Focus Unit, FFU/ml] was gained in VP-SFM containing FibraCel disks, and the lowest titer [3.66 +/- 0.4 _10[6] FFU/ml] was resulted on Cytodex-3 in M199. In these experiments, FibraCel disks supported the growth of Vero cells better than the microcarriers, and the use of DMEM for propagation of Vero cells and VP-SFM for proliferation of rabies virus showed better results. The experimental vaccine prepared by collected virus from VP-SFM has an acceptable potency of 2.75 IU. Based on these results and to the relative ease for making FibraCel disks, we recommend the use of this carrier for propagation of Vero cells and production of rabies virus
ABSTRACT
Multiple sclerosis and its murine model, experimental autoimmune encephalomyelitis [EAE], are chronic inflammatory demyelinating diseases of CNS. This study aimed to examine the effects of IL-27coding plasmid on disease status and certain immunological parameters in EAE-affected C57BL/6 mice. IL-27 gene was subcloned in P240 plasmid. The recombinant P240-mIL27 and P240 plasmids were injected two times, each time 200 micrograms, to test and control EAE mice, respectively. The clinical signs of the treated mice were evaluated daily and scored according to a standard method. One week after the last injection, all mice were sacrificed. The ELISA and MTT tests were performed to evaluate the production of IL-4, IFN-? and IL-17 from and proliferative response of splenocytes against specific antigen challenge, respectively. Furthermore, to demonstrate the immune cells infiltration, histopathological exam was performed on the brainstem of mice. The P240-mIL27 plasmid could significantly improve clinical course of EAE in the test group. Also in this group, the level of IL-4 was greater than that in the control group, while the levels of IFN-? and IL-17 were lower than those of the control group. In MTT test, the splenocytes of the test group showed a significantly less proliferative response than the control group. Finally, less infiltration of immune cells was seen in the brainstem of EAE mice treated with P240-mIL27 plasmid. IL-27 by shifting the immune responses from inflammatory Th1/Th17 towards anti-inflammatory Th2-type responses could be a suitable candidate for the treatment of inflammatory diseases such as MS