scholarly journals Construction of polycistronic baculovirus surface display vectors to express the PCV2 Cap(d41) protein and analysis of its immunogenicity in mice and swine

2020 ◽  
Vol 51 (1) ◽  
Author(s):  
Ya-Yi Chen ◽  
Wei-Chen Yang ◽  
Yu-Kang Chang ◽  
Chi-Young Wang ◽  
Wei-Ru Huang ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Surface Display ◽  
Recombinant Baculovirus ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Challenge Study ◽  
Ifn Γ ◽  
Cellular Immune

Abstract To increase expression levels of the PCV2 Cap(d41) protein, novel baculovirus surface display vectors with multiple expression cassettes were constructed to create recombinant baculoviruses BacSC-Cap(d41), BacDD-2Cap(d41), BacDD-3Cap(d41), and BacDD-4Cap(d41). Our results reveal that the recombinant baculovirus BacDD-4Cap(d41) was able to express the highest levels of Cap(d41) protein. Optimum conditions for expressing the PCV2 Cap(d41) protein were determined, and our results show that 107 of Sf-9 infected with the recombinant baculovirus BacDD-4Cap(d41) at an MOI of 5 for 3 days showed the highest level of protein expression. Mice immunized with the 4Cap(d41) vaccine which was prepared from the recombinant baculovirus-infected cells (107) elicited higher ELISA titers compared to the Cap (d41) vaccine. The 4Cap(d41) vaccine could elicit anti-PCV2 neutralizing antibodies and IFN-γ in mice, as confirmed by virus neutralization test and IFN-γ ELISA. Moreover, the swine lymphocyte proliferative responses indicated that the 4Cap(d41) vaccine was able to induce a clear cellular immune response. Flow cytometry analysis showed that the percentage of CD4+ T cells and CD4+/CD8+ ratio was increased significantly in SPF pigs immunized with the 4Cap(d41) vaccine. Importantly, the 4Cap(d41) vaccine induced an IFN-γ response, further confirming that its effect is through cellular immunity in SPF pigs. An in vivo challenge study revealed that the 4Cap(d41) and the commercial vaccine groups significantly reduce the viral load of vaccinated pigs as compared with the CE negative control group. Taken together, we have successfully developed a 4Cap(d41) vaccine that may be a potential subunit vaccine for preventing the disease associated with PCV2 infections.

scholarly journals Evaluation of Adjuvant Effectiveness of Alum-Propranolol Mixture on the Immunogenicity of Excreted/Secreted Antigens of Toxoplasma gondii RH Strain

2020 ◽  
Vol 11 (3) ◽  
pp. 570-577
Author(s):  
Elyar Meshkini ◽  
Arash Aminpour ◽  
Khosrow Hazrati Tappeh ◽  
Shahram Seyyedi ◽  
Meysam Shokri
Keyword(s):  
Toxoplasma Gondii ◽  
Immune Responses ◽  
Negative Control Group ◽  
Negative Control ◽  
Delayed Type Hypersensitivity ◽  
Control Group ◽  
Cellular Immune Responses ◽  
Ifn Γ ◽  
Cellular Immune ◽  
Phosphate Buffered Saline

Purpose: The introduction of novel adjuvants is an important step in attempts to develop a safe and more efficient vaccine. The present study was performed to determine whether the use of a mixed beta-adrenergic receptor antagonist propranolol (PRP) and aluminum (alum), as an adjuvant, have efficacy for Toxoplasma gondii (T. gondii) vaccine to induce protective immunity in a mouse model. Methods: Female BALB/c mice divided into five groups were immunized with excretory-secretory antigens (ESA) vaccine, alum-ESA vaccine, PRP-ESA vaccine, and alum-PRP ESA vaccine, as well as with phosphate buffered saline (PBS), as a negative control group. The immune responses were evaluated by lymphocyte proliferation assay for measuring delayed-type hypersensitivity (DTH) response and by cytokine assay for evaluating IFN-γ and IL-5 levels. The survival rate of mice in all groups was assessed during a three-week monitoring period after an intraperitoneal challenge with T. gondii tachyzoites. Results: The results showed that mice immunized with PRP, as an adjuvant, could secret a higher level of IFN-γ, which was significant in comparison to other groups. However, mice vaccinated with alum-precipitated ESA antigen had ability to produce an elevated level of IL-5 compared to other mouse groups (p≤0.05). Moreover, alum-PRP co-administration together with ESA vaccine resulted in the longer survival of mice. Conclusions: The findings of this study revealed that the combination of alum-PRP adjuvants and ESA vaccine of T. gondii elicits both humoral and cellular immune responses, which are comparable to either alum or PRP alone.

scholarly journals Antiproliferation Effects of Nanophytosome-Loaded Phenolic Compounds From Fruit of Juniperus Polycarpos Against Breast Cancer in Mice Model: Synthesis, Characterization and Therapeutic Effects

2021 ◽  
Author(s):  
Soheila Moeini ◽  
Ehsan Karimi ◽  
Ehsan Oskoueian
Keyword(s):  
Breast Cancer ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Therapeutic Effects ◽  
Mice Model ◽  
Fruit Extract ◽  
Phenolic Fraction ◽  
Juniperus Polycarpos

Abstract Background: This research was performed to synthesize nanophytosomes-loaded high phenolic fraction (HPF) from Juniperus polycarpos fruit extract and investigate its antiproliferation effects against breast cancer in mice model. Results: The nanophytosomes-loaded HPF from Juniperus polycarpos fruit extract was synthesized. The mice trial was conducted to determine the possible toxic effects of the synthesized nanophytosomes. The anticancer, pro-apoptotic, and antioxidative activities of the nanophytosomes were determined. The nanophytosomes-loaded HPF had a spherical structure with a size of 176 nm and a polydispersity index coefficient of 0.24. The in-vivo study manifested that nanophytosomes-loaded HPF significantly improved weight gain and food intake compared to the negative control group (p<0.05). The nanophytosomes-loaded HPF significantly enhanced the expression of bax (3.4-fold) and caspase-3 (2.7-fold) genes but reduced bcl2 (3.6-fold) gene expression in tumor cells. The average tumor size was significantly decreased in mice treated with nanophytosomes-loaded HPF (p<0.05). The expression of GPX (2.3-fold) and SOD (2.7-fold) antioxidants in the liver of mice supplemented with nanophytosomes-loaded HPF was significantly developed compared to the negative control (p<0.05). The nanophytosomes-loaded HPF did not show toxicity on normal cells. Conclusion: Our results indicated that nanophytosomes-loaded HPF might be a potential anticancer agent for the breast cancer treatment.

scholarly journals The influence of whole sonicated Pseudomonas aeruginosa antigens on experimental p. aeruginosa arthritis in rabbits

2014 ◽  
Vol 38 (1) ◽  
pp. 1-10
Author(s):  
Ahmed Q. Al-Awadi
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Inflammatory Reaction ◽  
Rabbit Model ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Post Inoculation ◽  
Bacterial Isolation ◽  
Internal Organs ◽  
Ifn Γ

To study the influence of whole sonicated Pseudomonas aeruginosa antigens (WSPAgs) on experimental arthritis induced by this bacteria, 15 rabbits were divided into 3 equal groups. The 1st group was inoculated intraarticular with 0.2 ml of p. aeruginosa suspension (2×108 cfu/ml), the 2nd group was immunized with WSP Ags, and inoculated intraarticular as in the 1st group. The 3rd group was served as negative control group. At 30 day post inoculation the immunized (2nd) group showed increase in the cellular (DTH and IFN-γ) and humeral (IgG) immunity and moderate bacterial isolation from joints, blood and internal organs comparing with other groups. The 1st group showed sever symptoms and inflammatory reaction as well as very obvious gross and microscopical lesions in their joints including supportive reaction, pyogranulomatous lesions, necrosis, pannus reaction and destruction of the articular cartilage and the lesion extended to the subchondral bone leading to osteomyelitis, the 2nd group (immunized group) expressed mild to moderate inflammatory reaction and the microscopic examination indicate that the lesion was confined in the articular capsule. In conclusion the whole Pseudononas aeruginosa sonicated Ags (WSPAgs) protect the joint from the experimental infection by P. aeruginosa in a rabbit model.

scholarly journals THE PROTECTIVE ROLE OF OMEGA-3 AGAINST GENOTOXICITY AND REPRODUCTIVE TOXICITY OF COBALT OXIDE NANOPARTICLES ACUTE TREATMENT IN MALE MICE

2018 ◽  
Vol 11 (5) ◽  
pp. 423
Author(s):  
Nahed A Hussien ◽  
Hanan R. H. Mohamed
Keyword(s):  
Cobalt Oxide ◽  
Negative Control Group ◽  
Protective Role ◽  
Negative Control ◽  
Control Group ◽  
Omega 3 ◽  
Genotoxic Potential ◽  
Polychromatic Erythrocytes

Objective: Cobalt nanoparticles (NPs), especially cobalt oxide NPs (Co3O4 NPs) are attracting unique shaped NPs that are used in different biomedical applications and medicine. Different in vitro studies report their toxic and carcinogenic effect but limited in vivo studies were present on its genotoxic potential. The present study was aimed to evaluate the genotoxic potential of Co3O4 NPs on bone marrow cells and sperms and the protective role of omega-3 in male albino mice.Methods: Animals were segregated into four groups that were orally treated for 3 consecutive days, Group 1: Negative control; Group 2: Omega-3 (250 mg/kg); Group 3: Co3O4 NPs (20 mg/kg); and Group 4: Combined group (250 mg/kg Omega-3 and Co3O4 NPs 20 mg/kg).Results: The present results show that Co3O4 NPs administration significantly increased number of micronucleated polychromatic erythrocytes (PCEs)/1000 PCEs, sperm abnormalities, and DNA damage, significantly decreased sperm motility and concentration in comparison to negative control group. However, Omega-3 administration in the combined group modulates the genotoxic potential of Co3O4 NPs in comparison to Co3O4 NPs group.Conclusion: The present study reports the genotoxic potential of Co3O4 NPs in vivo and assesses the protective role of Omega-3 administration due to its antioxidant effect.

dSLIM Immunomodulators Induce Anti-Tumor Responses Both In Vitro and In Vivo.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1727-1727
Author(s):  
Manuel Schmidt ◽  
Javier de Cristobal ◽  
Astrid Sander ◽  
Bernadette Brzezicha ◽  
Sven A. König Merediz ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Tumor Cells ◽  
Neutralizing Antibodies ◽  
De Novo ◽  
Tumor Cell Line ◽  
Control Group ◽  
Γδt Cells ◽  
Ifn Γ

Abstract Cytosine-guanine (CpG) motifs containing oligonucleotides (ODN) are commonly used for immunomodulatory purpose in cancer therapy and for the treatment of allergic diseases since they resemble bacterial DNA and serve as “danger signals”. These CpG-ODNs promote predominately a TH1-response with secretion of IL-12 and IFN-γ, In addition their broad potential includes activation of B-cell proliferation, monocyte stimulation and secretion of IgM and IL-6, and stimulation of plasmacytoid DC to produce IFN-α/-β and thus γδT-cells and NK-cells to express CD69 and secrete IFN-γ. Usually phosphorothioate (PS) modifications are to enhance the stability, but these are leading to several side-effects, like severe organ enlargements, morphological changes and immunosuppression in mice. We designed immunomodulatory molecules based on short covalently-closed dumbbell-like structures (dSLIM) to stabilize the DNA without the otherwise necessary PS-modification. To evaluate the anti-tumor effect of the dSLIM molecules we developed an in vitro anti-tumor assay. This assay uses supernatant from dSLIM-activated human PBMCs for incubation with tumor cells in vitro. We observed increased apoptosis and necrosis of the HT-29 tumor cell line after incubation with supernatant from dSLIM-treated PBMC which was significantly higher than the effect of supernatant from non-treated PBMC. In addition, supernatant from dSLIM-treated PBMC increased the expression of HLA-ABC on the tumor cells, a pre-requisite for tumor cell recognition by the immune system. These effects were confirmed with human HEK293 and murine Renca cell lines. Analyzing the effect with neutralizing antibodies to various apoptosis-related cytokines, we observed a crucial role of IFN-γ but not IFN-α or TNFα. To investigate the anti-tumor effects of dSLIM in vivo, we employed a SKH1 murine model which is prone to spontaneous development of papillomas. Using chemicals for initiation and weekly promotion of de novo papilloma development we compared groups of weekly s.c. or i.p. dSLIM injections, respectively, with the PBS control group. The number of papilloma developing mice was significantly lower in the dSLIM groups and the total number of papillomas on all mice was reduced by approximately 50%. In conclusion, we showed that dSLIM immunomodulators exhibit potent anti-tumor effects in vitro and in vivo.

scholarly journals BacMam virus-based surface display for HCV E2 glycoprotein induces strong cross-neutralizing antibodies and cellular immune responses in vaccinated mice

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Ebrahim Kord ◽  
Farzin Roohvand ◽  
Jean Dubuisson ◽  
Thibaut Vausselin ◽  
Hosein Nasr Azadani ◽  
...  
Keyword(s):  
Immune Responses ◽  
Mammalian Cells ◽  
Neutralizing Antibodies ◽  
Surface Display ◽  
Immunogold Electron Microscopy ◽  
Cellular Immune Responses ◽  
Boost Immunization ◽  
Ifn Γ ◽  
Cellular Immune ◽  
And Control

Abstract Background Despite recent advancements, limitations in the treatment and control of hepatitis C virus (HCV) infection reprioritized the studies for invention of an efficient HCV vaccine to elicit strong neutralizing antibodies (NAbs) and cellular responses. Methods Herein, we report molecular construction of a BacMam virus-based surface display for a subtype-1a HCV gpE2 (Bac-CMV-E2-gp64; Bac) that both expressed and displayed gpE2 in mammalian cells and bacouloviral envelope, respectively. Results Assessments by western blotting, Immunofluorescence and Immunogold-electron microscopy indicated the proper expression and incorporation in insect cell and baculovirus envelope, respectively. Mice immunized in three different prime-boost immunization groups of: Bac/Bac, Bac/Pro (bacoulovirus-derived gpE2) and Bac/DNA (plasmid DNA (pCDNA)-encoding gpE2) developed high levels of IgG and IFN-γ (highest for Bac/Bac group) indicating the induction of both humeral and cellular immune responses. Calculation of the IgG2a/IgG1 and IFN-γ/IL-4 ratios indicated a Th1 polarization of immune responses in the Bac/Bac and Bac/DNA groups but a balanced Th1-Th2 phenotype in the Bac/Pro group. Sera of the mice in the Bac/Bac group provided the highest percentage of cross-NAbs against a subtype-2a HCVcc (JFH1) compared to Bac/Pro and Bac/DNA groups (62% versus 41% and 6%). Conclusions Results indicated that BacMam virus-based surface display for gpE2 might act as both subunit and DNA vaccine and offers a promising strategy for development of HCV vaccine for concurrent induction of strong humoral and cellular immune responses.

INFLUENCE OF JAMU MADURA “EMPOT SUPER” ON THE VAGINAL EPITHELIUM THICKNESS OF WHITE MICE (Rattus norvegicus) – AN IN VIVO STUDY

2017 ◽  
Vol 2 (1) ◽  
pp. 47
Author(s):  
Anik Listiyana
Keyword(s):  
Rattus Norvegicus ◽  
Negative Control Group ◽  
Negative Control ◽  
Vaginal Epithelium ◽  
In Vivo Study ◽  
Control Group ◽  
Control Group Design ◽  
Post Test ◽  
Epithelium Thickness

<p><em>The aim of this research is to determine the influence of jamu Madura “Empot Super” (JMES) on the vaginal epithelium thickness of Rattus norvegicus in vivo. This research is kind of “true experimental-post test only control group design”. The rats were given drinking JMES once daily PS (Per-Sonde) for a month, then the vagina was taken to be sample for HE colouring. The sample was observed by the binocular microscope (100 times magnification) to identify the changes in the thickness of their vaginal epithelium. Calculation of the vaginal epithelium thickness was counted on the 10 field of view chosen randomly by the blind method. The result show that the vaginal epithelium thickness increased with dose 0,17mg/BW, 0,34mg/BW, and 0,68mg/BW of JMES compared with negative control group. But, the vaginal epithelium thickness decrease at the dose 0,51mg/BW compared with negative control group.</em></p><p> </p><p><strong>Keywords</strong><strong>: </strong>Jamu Madura “Empot Super” (JMES), vaginal epithelium thickness, white mice (<em>Rattus norvegicus</em>), In Vivo study</p>

In vivo Antitumoral Effect of Plantago major L. Extract on Balb/C Mouse with Ehrlich Ascites Tumor

2007 ◽  
Vol 35 (05) ◽  
pp. 841-851 ◽  
Author(s):  
Mehmet Ozaslan ◽  
I. Didem Karagöz ◽  
M. Emin Kalender ◽  
I. Halil. Kilic ◽  
Ibrahim Sari ◽  
...  
Keyword(s):  
Weight Gain ◽  
Negative Control Group ◽  
Negative Control ◽  
Ehrlich Ascites Tumor ◽  
Control Group ◽  
Plantago Major ◽  
Ascites Tumor ◽  
Treatment Groups ◽  
Ehrlich Ascites

The aim of this study is to investigate the antitumor activity of Plantago major L. extract in Ehrlich ascites tumor (EAT) bearing Balb/C mice in vivo. Thirty male Balb/C mice were divided into 5 groups: 3 treatment groups and 2 control groups (6 per group). Treatment groups and the negative control group were injected with EAT (1 × 106 cells) intraperitoneally to develop ascites tumor. P. major L. extract (1%, 2% and 3% concentration extracts, 0.1 ml/day/mouse) were given p.o. for 10 alternate days. The control group was treated with 0.9% NaCl solution (0.1 ml/day/mouse). The changes of body weight in animals were recorded. On the 11th day, all of the mice were sacrified and their tissues were stained with haematoxylen and eosin for pathological studies. Body weights of in 3 treatment groups and the negative control group were elevated because of tumor burden. The maximal weight gain was recorded in the negative control group and the minimal weight gain was recorded in Group I. Pathological studies showed that P. major L. extract (especially 1% concentration) has inhibitive effect on EAT. P. major has an inhibitory effect on EAT in a dose dependent manner.

scholarly journals Preparation and Evaluation of Azelaic Acid Topical Microemulsion Formulation: In Vitro and In Vivo Study

Pharmaceutics ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 410
Author(s):  
Wan-Hsuan Hung ◽  
Ping-Kang Chen ◽  
Chih-Wun Fang ◽  
Ying-Chi Lin ◽  
Pao-Chu Wu
Keyword(s):  
Azelaic Acid ◽  
Skin Irritation ◽  
Topical Administration ◽  
Negative Control Group ◽  
Negative Control ◽  
Commercial Product ◽  
Control Group ◽  
Drug Delivery Carrier

The aim of this study was to design oil in water (O/W) microemulsion formulations for the topical administration of azelaic acid. The permeability of azelaic acid through rat skin and the anti-inflammatory activities of the formulations were conducted to examine the efficacy of the designed formulations. Skin irritation and stability tests were also performed. The permeability of azelaic acid was significantly increased by using O/W microemulsions as carriers. The edema index of ear swelling percentage was significantly recovered by the 5% drug-loaded formulation and a 20% commercial product, demonstrating that the experimental formulation possessed comparable effect with the commercial product on the improvement of inflammation. The experimental formulation did not cause significant skin irritation compared to the negative control group. Moreover, the drug-loaded formulation also showed thermodynamic stability and chemical stability after storage for 30 days. In conclusion, the O/W microemulsion was a potential drug delivery carrier for azelaic acid topical application.

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