| As the second most common malignancy in women worldwide, cervical cancer badly threatens the health of female. Previous studies have shown that it is closely correlated with the infection of Human papillomaviruses (HPV). Thus prevention and treatment of cervical cancer can be achieved through control of HPV infection. Recently, prophylactic vaccine of HPV has been presented abroad. HPV16L2E6E7 (HPV vaccine) is a fusion protein vaccine that is being developed in China, and now application of clinical trial has been in progress. However, like other protein vaccine, it still has disadvantage of poor immunogenicity. Addition of adjuvants in vaccine is commonly used to enhance immune response. This study was designed to evaluate the immunological effect of docetaxel and ginsensiode on HPV vaccine so as to provide references for clinical application.1 Adjuvant effects of docetaxel in miceObjective:To determine if docetaxel has immunoadjuvant effects for HPV vaccine. Methods: A total of 2 experiments were carried out and described as follows. In experiment A, C57BL/6 mice were subcutaneously (s.c.) challenged with 104TC-1 cells (100 μl) into the right groin on day-1, and then intramuscularly (i.m.) injected with saline,200 μg docetaxel or 120 μg of HPV vaccine in 200 μl of saline solution containing docetaxel (0,100,200 or 400 μg) at hind limbs on days 0,3 and 7. On day 21, blood samples were collected for determination of IgG and the isotypes responses; splenocytes were harvested for analysis of CTL activity, IFN-y and IL-10 secreting cells, Treg cells; tumor nodules were collected for measurement of the weight and volume as calculated by length/3 x width x height. In experiment B, the immunization and groups were carried out and described as experiment A. Results:When TC-1 tumor-bearing mice were injected with HPV vaccine dissolved in the solution containing docetaxel, significantly less volume and weight of the tumors, and more percent survival were detected than those treated with HPV vaccine alone. The inhibition of tumors was associated with significantly activated CTLs, increased IFN-y secreting T cells, serum antigen-specific IgG and the isotypes as well as suppressed Treg cells and IL-10 secreting cells.2 Adjuvant effects of Ginsenoside Rgl in miceObjective:To determine if Ginsenoside Rgl has immunoadjuvant effects for HPV vaccine. Methods:C57BL/6 mice were intramuscularly (i.m.) injected with saline,100 μg of HPV vaccine, or 10 μg of HPV vaccine in 200 μof saline solution containing Rg1 (0,25,50,100 or 200 μg) at hind limbs on days 0,3 and 7. On day 21, blood samples were collected for determination of IgG and the isotypes responses; splenocytes were harvested for analysis of lymphocyte proliferation responses, mRNA expression of IL-4 and IL-12, IFN-γ secreting T cells. Results:Rg1 augmented significantly higher responses of serum specific IgG and IgG isotypes responses as well the number of IFN-γ secreting cell, lymphocytes proliferation as mRNA Expression of IL-4 and IL-10 than immunization of HPV in mice.3 Adjuvant effects of Ginsenoside Rg3 in miceObjective:To determine if Ginsenoside Rg3 has immunoadjuvant effects for HPV16 vaccine. Methods:Mice were intramuscularly (i.m.) injected with saline or 120μg of HPV vaccine in 200 μl of saline solution containing Rg3 (0,25,50 or 100 μg) at hind limbs on days 0,3 and 7.On day 21, blood samples were collected for determination of IgG; splenocytes were harvested for analysis of IFN-γ secreting T cells. Results:Rg3 augmented significantly higher responses of serum specific IgG and the number of increased IFN-γ secreting T cell.4 Adjuvant effects of the formulation of Rg3 and docetaxel in miceObjective:To determine if the formulation of Rg3 and docetaxel has immunoadjuvant effects for HPV vaccine. Methods:A total of 2 experiments were carried out and described as follows. In experiment A, mice were subcutaneously (s.c.) challenged with 104TC-1 cells (100μl) into the right groin on day-1, and then intramuscularly (i.m.) injected with 200μl of saline with (0,200 μg of docetaxel,100 μg of Rg3, or the formulation including 200 μg of docetaxel and 100 μg of Rg3) or 120 μg of HPV vaccine in 200 μl of sanline with (0,200 μg of docetaxel,100 μg of Rg3 or the formulation containing 200 μg of docetaxel and 100 μg of Rg3) at left (100 μl) and right (100 μl) hind limbs on days 0,3 and 7.On day 21, blood samples were collected for determination of IgG; splenocytes were harvested from sacrificed mice for IFN-γ secreting T cells; tumor nodules were collected for measurement of the weight and volume as calculated by length/3×width×height. In experiment B, the immunization and groups were carried out and described as experiment A. To observe survival of animals, mice were monitored daily until day 60. Results:When TC-1 tumor-bearing mice were injected with HPV vaccine dissolved in the solution containing docetaxel and Rg3, less volume and weight of the tumors, more percent survival and the number of IFN-γ secreting T cell, higher responses of serum specific IgG were detected, but there was no deference between the formulation of Rg3 and docetaxel and docetaxel only.5 Effects of docetaxel on cell mediated immunityObjective:To determine if docetaxel can improve cell mediated immunity. Methods:C57BL/6 mice intramuscularly (i.m.) injected with 120 μg of HPV vaccine in hind limbs on days 0,3 and 7. On day 21, splenocytes were harvested from sacrificed mice for analysis of CTL activity and mRNA express of Fas、FasL、M6PR、perforin、GzB and caspase-3. Results:Docetaxel can enhance CTL cytotoxicity against TC-1 cells. It was referred to increase of mRNA expression of Fas, M6PR, Caspase-3, FasL, GzB gene by docetaxel.6 Effects of docetaxel on tumor microenvironmentObjective:To determine if docetaxel can improve the tumor microenvironment. Methods: C57BL/6 mice were subcutaneously (s.c.) challenged with 104 TC-1 cells (100 μl) into the right groin on day-1, and then intramuscularly (i.m.) injected with saline,200 μg docetaxel or 120 μg of HPV vaccine in 200 μl of saline solution containing docetaxel (0,200 μg) at hind limbs on days 0,3 and 7. On day 21, Mononuclear cells were isolated from tumor nodules for analysis of Treg cells and IL-10, IFN-y, VEGF and STAT-3 mRNA levels. Results:Docetaxel can decrease Treg cells, suppressed IL-10, VEGF and STAT-3 mRNA levels and upregurated IFN-y mRNA levels in tumors.In conclusion, docetaxel could significantly strengthen the anti-tumor ability of HPV vaccine and length the survival time in tumor-bearing mice. Rgl and Rg3 have immunoadjuvant effects for HPV vaccine. However, the immune effect was not increased when it was the formulation of Rg3 and docetaxel compared with the latter was used alone in tumor bearing mice; The adjuvant effect of docetaxel may be due to enhance cell mediated immunity by the increasing of GzB, M6PR, Fas, FasL and caspase-3 mRNA expression and improve the tumor microenvironment by deceased Treg cells, suppressed IL-10, VEGF and STAT-3 mRNA levels and upregurated IFN-y mRNA levels in tumors. The study has important significance for using docetaxel and ginsenoside as adjuvant to enhance the immune effect of HPV vaccine. |
PDF Full Text Request