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العنوان
The Possible Effect of Metal Nanoparticles
Associated with Cancer Cells Vaccines Formulated
by Gamma Rays to Initiate an Antitumor
Immunomodulatory Response /
المؤلف
Shams El-din, Rokaya Elsayed Maarouf Elsayed.
هيئة الاعداد
باحث / Rokaya Elsayed Maarouf Elsayed Shams El-din
مشرف / Amina M. Medhat
مشرف / Neamat H. Ahmed
مناقش / Ahmed I. El-Batal
تاريخ النشر
2019.
عدد الصفحات
251 P. :
اللغة
الإنجليزية
الدرجة
الدكتوراه
التخصص
Biochemistry
تاريخ الإجازة
1/1/2019
مكان الإجازة
جامعة عين شمس - كلية العلوم - قسم الكيمياء الحيوية
الفهرس
Only 14 pages are availabe for public view

from 251

from 251

Abstract

Cancer vaccines are considered as active immunotherapy as they boost the body’s immune system to defend against cancer. Vaccines containing preparations of whole tumor cells or tumor lysate have the advantage of simultaneously presenting multiple defined and undefined TAAs to the immune system (De Gruijl et al., 2008), thereby expanding the repertoire of both CD4+ (helper) and CD8+ (cytotoxic) T lymphocytes that are activated, this consequently decreases the possibility of immune evasion by the tumor.
When nanomedicines are applied in vivo, they act as foreign materials and induce the immune response, immunosuppression or immunostimulation (Dobrovolskaia and McNeil, 2007). NPs play as a Janus’ double-face in nanomedicine applications. On one hand, the effects to the Immune system may be neit treatment of disease through enhancing immune response. On the other hand, the immunomodulation of NPs may bring harm (Parween et al., 2011).
In the current study; we sought to evaluate the impact of whole tumor cell vaccine, tumor cell lysate vaccine each alone or combined with silver nanoparticles against Ehrlich carcinoma in mice. Silver nitrate (1mM) and polyvinyl pyrrolidone (PVP) solutions were used for the synthesis of silver nanoparticles (Du et al., 2008) and 30.0 kGy gamma radiation at room temperature (Gasaymeh et al., 2010 and El-Batal et al., 2013). After irradiation, the size, morphology and stability of the synthesized AgNPs were characterized using UV-Visible absorption Spectroscopy, fourier transform infrared (FTIR), dynamic light scattering technique (DLS), transmission electron microscopy (TEM) and X-ray study. Tumor cell lysate vaccine was prepared by irradiation of Ehrilch cells at 8 kGy gamma irradiation (Salama and Hassan, 2014) and lysis according to the method of (Scchnurr et al., 2001) with modification (Deutsch et al., 2000).
Whole cell vaccine was prepared by irradiating the cells with 3000 rads (Srikanth et al., 2002). Solid Ehrlich carcinoma was induced by intramuscular inoculation in the left thigh of the lower limb of each mouse with 0.2 ml of EAC cells (Fahim et al., 1997). At next day after tumor induction, mice were injected with 0.1ml of AgNPs (500 µg/ml concentration) intratumoraly at left thigh daily from day 2 to day 5 and a booster dose on 12th day was also administered.
Four doses of whole cell vaccine were injected intraperitonealy at the rate of 100 µl /10 gm body weight of the animal daily from day 2 to day 5. After 7 days’ interval, on 12th day a booster dose of the same strength was administered. Tumor cell lysate vaccine was injected in the right thigh one time /week for 3 successive weeks then after 14 days from the last injection, ehrilch tumor was injected.
In vivo experiments were performed on healthy and ehrlich carcinoma bearing female swiss albino mice. Mice were randomly assigned and equally divided into seven experimental groups of 10 mice each as follows: group (1): normal control group (NC); group (2): Ehrlich group; group (3): Ehrlich carcinoma bearing mice treated with AgNPs (ECAg) group; group (4): Ehrlich carcinoma bearing mice treated with whole cell vaccine (ECWCV) group; group (5): Ehrlich carcinoma bearing mice treated by both silver nanoparticles and whole cell vaccine (ECAgWCV); group (6): Ehrlich carcinoma bearing mice vaccinated with lysate (ECLys) and group (7): Ehrlich carcinoma bearing, lysate vaccinated, AgNPs treated group (EcAgLys).The growth rate of solid tumor in all experimental groups was measured by Caliper.
At the end of the experiment, the whole blood was withdrawn from animals and was centrifuged and the upper layer (serum) was taken for determination of serum nitric oxide level, the activity of catalase enzyme, serum IFN-γ and IL-10 levels by ELISA technique. In addition, CD4+T and CD8+T count, TGF-β level, P53 activity and caspase-3 activity were determined in tumor tissue by flow cytometry analysis. Also, histopathological examination, apoptosis and necrosis of tumor, lymph node and spleen tissues were performed to evaluate the histopathological and apoptotic changes.
The results obtained were statistically analyzed and can be summarized as follows:
• EAC cells treated in vitro with different concentrations of silver nanoparticles, whole cell vaccine or tumor cell lysate vaccine showed significant cell inhibition.
• Significant inhibition in tumor size was observed through the observation period in groups of experimental animals treated with whole cell vaccine or vaccinated with tumor cell lysate vaccine, each alone or combined with silver nanoparticles.
• Significant decrease in serum nitric oxide level and increase in serum catalase activity was observed after treatment of tumor bearing mice with whole cell vaccine, vaccination with tumor cell lysate vaccine, each alone or combined with AgNPs.
• Treatment of EC bearing mice with WCV or vaccination with tumor cell lysate vaccine before tumor induction, each alone or combined with AgNPs treatment, caused reduction in CD4+ population and increase in CD8+population compared to EC group.
• WCV, tumor cell lysate vaccine either each alone or combined with AgNPs cause reducing IFN-γ level and increasing IL-10 protein level in comparison with EC group.
• Decrease in TGF-ß level, decrease in P53 level and elevation of caspase-3 activity, was observed after treatment of tumor bearing mice with WCV, vaccination with cell lysate vaccine before tumor induction, each alone or combined with AgNPs with respect to EC group.
• Apoptotic and histological findings in tissues (tumor, lymph node and spleen) showed an antitumor effect of whole tumor cell vaccine, tumor cell lysate vaccine, each alone or combined with silver nanoparticles.
• According to the obtained results, it could be concluded that tumor cell lysate vaccine and whole cell vaccine each alone or combined with silver nanoparticles exhibited significant antitumor activity and that immunization with the cell lysate vaccine acts more efficiently as an immunomodulator especially when combined with silver nanoparticles.