الفهرس | Only 14 pages are availabe for public view |
Abstract Cancer is a major cause of death worldwide. Radiotherapy and chemotherapy also affect normal cells and results in side effects that limit treatment. The rapid increase in knowledge of the immune system and its regulation has led to a resurgence of interest in immunological approaches as a target to eliminate cancer. Xenogeneic placental protein extract sterilized either using 0.2 micron Millipore filters (Centricon) or 2Gy gamma irradiation and injected in mice as a vaccine stimulus to the immune system before challenge with viable Ehrlich tumor. This study aims to investigate the efficiency of the prepared vaccine in immunotherapy against tumor cells. To fulfil this aim, the MTT viability test which was assayed as a cytotoxicity assay by using spleen cells, tumor size, animal weight was also measured, lymphocyte count, CD8, granzyme B, caspase-3, (MMP-2 & MMP-9) activities and protein fractionation were investigated in blood of the different studied groups.In order to fulfil the target of this study, a total of 255 female swiss albino mice weighing 22-25g were used. They were divided into four groups, G1: is the untreated group (control group contains 15 mice), G2: (120 mice) is the Ehrlich treated group (injected with 2.5×105 viable Ehrlich tumor cells/mouse). Third group (G3): is vaccinated with 6 mg/mL xenogeneic placental extract sterilized using 0.2 micron Millipore filters. Fourth group (G4): is vaccinated with 6 mg/mL xenogeneic placental extract sterilized using Gamma irradiation at dose level 2 Gy. Each of the third and fourth group contains 60 mice and divided into 4 subgroups (a, b, c and d) each contains 15 mice. Results obtained from this study demonstrated high significant decrease in tumor size for G3a, G3c, G3d, G4a, G4b, G4c & G4d subgroups, Which was detected 10 days after tumor appearance in Ehrlich treated (G2) group. The animal weight in G3a, G3b, G3d, G4a, G4b, G4c & G4d subgroups, showed significant increase compared to G2 group.Lymphocyte count: This study recorded a significant increase in lymphocyte count in G3d, G4b, G4c and G4d subgroups compared to G1 group. On the other hand, G3d, G4a, G4b, G4c and G4d subgroups showed significant increase in lymphocyte count compared to G2 group. CD8 activity in blood: The results of this study revealed that there was significant increase in G4d subgroup compared to G1group. On the other hand, G4b and G4d subgroups showed significant increase in lymphocyte count compared to G2 group. Granzyme B activity in blood: The present study showed that the granzyme B activity showed significant increase in G4d subgroup compared to G1 and G2 groups. Caspase-3 activity: The study showed a significant increase in caspase-3 activity for G4c and G4d subgroups compared to G1 and G2 subgroups.Serum MMP-(2&9) activities: Gelatin zymography for serum MMP-(2&9) activity (70&90KDa) in all the studied groups demonstrated that MMP2 and MMP9 bands were detected in all the studied groups but with less pronounced bands than control group. The G4d subgroup displays the less pronounced band compared to the other studied groups and to control group. Protein fractionation: Gel electrophoresis for protein fractionation of Ehrlich ascites protein, placental protein extract and in serum of all the studied groups showed that Ehrlich ascites protein (lane 2) and serum sample of placental protein extract (lane 3) are represented by four bands. The first was observed at 7KDa, suggesting that it is the migration inducing gene-7 (Mig-7). A second band was detected between the 29KDa and 45kDa suggesting that it is for trophoblast-derived growth factor. A third band was detected at >67KDa suggesting that it is for integrin and a fourth band was observed at >116KDa suggesting that it is for the epidermal growth factor receptor (EGFR). |