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Brucella is a facultative intracellular pathogen, and brucellosis is commonest zoonotic disease worldwide. Brucella species, isolated from domestic animals, are important pathogen for humans. Annually, more than 500,000 new cases of brucellosis are reported, and this figure is an underestimate due to extended under-reporting cases in several endemic countries. Brucella has a variety of virulence mechanisms that prevent detection and activation of innate immunity, but protection against intracellular pathogen is represented by cell-mediated immunity. As yet, much research has been performed to develop a safe Brucella vaccine to control the disease in human and animals. Despite the availability of several live attenuated vaccine for animals, currently, no effective human vaccine is available. Moreover, due to the potential use of Brucella in bioterrorism or biowarfare, development of an effective vaccine against brucellosis for human use is necessary. In this paper, we aimed to review and discuss the efforts of researchers to develop vaccines against Brucellosis.

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Background: Group B streptococcus (GBS) is the major cause of serious life threatening infections in neonates, pregnant women, and other adults with underlying diseases. Capsular polysaccharide typing is a significant way for epidemiological studies of GBS, the pathogenesis, and other studies associated with GBS infections including surveillance programs and vaccine development in future. Molecular serotyping (MS) methods offer more accurate and reliable typing of bacteria. The aim of current study was to differentiate genotypes of clinical GBS isolates based on PCR assay to acquire information about the distribution of GBS types in Hamadan, Iran.
Materials and Methods: A total of 62 clinical GBS strains including vaginal swabs, urine cultures, and blood culture isolates were examined for genotyping using multiplex PCR assay.
Results:Among the 62 GBS isolates examined, all capsular types, except VI, VII, and VIII, were found. Type III was the predominant type with 35 isolates (56.5%), followed by Type V with 11 isolates (17.7%), Type II with 7 isolates (11.3%), Type Ia with 5 isolates (8.1%), and Types Ib and IV with similar prevalence of 2 isolates (3.2%) for each type.
Conclusion: The results of the current study demonstrated that Type III is the predominant type in Hamadan, followed by Types V, II, Ia, Ib, and IV, respectively. Using MS method leads to accurate, sensitive, specific, and fast typing of GBS isolates. The advantages of MS method allow it to analyze various populations and to examine invasive and colonizing isolates in extensive epidemiological studies and surveillance activities. In fact, MS will facilitate the proper formulation of candidate GBS vaccines.

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Newcastle disease is a fatal viral disease which is highly contagious that affects most species of birds and is a major economic threat in the poultry industry. Both the HN and F glycoproteins of Newcastle disease virus (NDV) are essential for pathogenicity and virus infectivity. This study describes immunization of DNA vaccines encoding the HN, F or both the genes of New castle disease virus. In our previous study, the antigen expression of the insert genes has been validated in vitro by Western Blotting and Indirect Immunosenest. In this study, ELISA and HI analysis of the in vivo experiment on SPF (specific Pathogen Free) chickens showed the induction of humoral responses by the DNA vaccines. Our finding indicated that twice vaccination with pDNA was able to elicit significant antibody titers (P< 0.05) by either monocistronic (pIRES/HN and pIRES/F) or bicistronic (pIRES/F/HN) plasmid, after one week of second pDNA vaccination (booster). The results proposed that DNA immunization of chickens at second vaccination had enhanced the antibody response successfully. Also, it revealed that vaccination with the co-expression plasmid pIRES/HN/F can induce a stronger antibody response than vaccination with pIRES/HN or pIRES/F alone.

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Backgrounds: A short sequence of viral protein or peptide, can be used as a potential vaccine for the treatment of that virus. Considering all variants of concern (VOC), vaccine design with peptide for Severe Acute Respiratory Syndrome coronavirus 2 (SARS CoV2) is a challenging job for scientists.
Materials & Methods: In this current study, an epitope containing peptide vaccine for nonstructural protein 4 (nsp 4) of SARS CoV2 coronavirus has been predicted. With the help of a modified method for both B and T call epitope prediction, verified by molecular docking studies, linear B cell and T cell epitopes for nsp4 protein, are predicted here. Predicted epitopes are analyzed further with population coverage calculation and epitope conservancy analysis.
Findings: A short peptide sequence   ⁷⁴QRGGSYTNDKA⁸⁴  has been selected as B cell epitope considering the scores for surface accessibility, hydrophilicity, beta turn prediction for each amino acid residues.
Similarly, the peptide sequences ³⁵⁹ FLAHIQWMV³⁶⁷ and   ³⁵⁹ FLAHIQWVMFTPLV³⁷³ are predicted as T cell epitopes for MHC-I and MHC-II molecules. These two potential epitopes can interest with HLA-A*02:01 and HLA-DRB*01:01, MHC allelic proteins respectively with lowest IC₅₀ values.
Furthermore, no amino acid mutations are observed in GISAD Global initiate on sharing all influenza data) database for alpha, beta, gamma and delta variance of concerns (VOC). Among seven amino acid point mutation of nsp 4 protein in Omicron variant, none of them is present in the peptide sequences of predicted epitope-based vaccines.
Conclusion: The short peptide sequences can be predicted as vaccines to prevent coronavirus infections for all variants of concerns.
 

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Backgrounds:  Although conventional therapies have played an essential role in the treatment of many diseases, emerging diseases require new treatment methods with less complications. Therefore, it is important to develop an effective vaccine for infections caused by the coronavirus to prevent mortality and create immunity the community.
Materials & Methods: In this research bioinformatics tools were used to design a vaccine against the  M membrane protein of SARS-CoV-2.  A total of 27 epitopes confined to B cells and MHC I and II alleles were structurally constructed in M protein for immune stimulation and antibody recognition which were used in the construction of a chimeric peptide vaccine .
Results: The vaccine was predicted to be a stable, antigenic, and non-allergenic compound. TRL5/vaccine complex  analysis  and docking simulation indicated a sufficiently stable binding with appropriated receptor activation. The immune response simulation following hypothetical immunization indicated the potential of this vaccine to stimulate the production of active and memory B cells, CD8 + T and, CD4 + T cells, and effective immunological responses induced by Th2 and Th1.
Conclusion:  The analysis of in-silico processes showed that the vaccine structure induced high antigenicity and good cellular immunity in the host body and stimulates various immune receptors such as TLR5, MHC I, and MHC II. Vaccine function was also associated with an increase in IgM and IgG antibodies and a set of Th1 and Th2 cytokines. But the final confirmation of the effectiveness of the designed vaccine requires  clinical processes.

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Backgrounds: Several studies have elucidated vitamin D as an important immunomodulatory factor regulating immune responses to different viral infections and vaccines. This study aimed to evaluate the impact of 25(OH) D serum levels on immune responses to hepatitis B virus (HBV) vaccine.
Materials & Methods: This study was conducted on 134 healthy individuals aged 18-35 years, referring to health centers for HBV vaccination in Mane and Samalghan city in North Khorasan, Iran from June to September 2021. Demographic data were collected through a questionnaire. Serum 25(OH) D levels were analyzed using commercial sandwich ELISA kits. Anti-hepatitis B surface antibody (anti-HBsAb) levels were determined in blood samples 4-6 weeks post-vaccination.
Findings: The prevalence of vitamin D deficiency and insufficiency among the participants was 46.3 was 34.3%, respectively. The level of 25(OH) D was insignificantly higher in women than in men. There was no significant association between serum 25(OH) D levels and participants' ethnicities and BMI ranges. Anti-HBsAb titer was significantly higher in participants with sufficient vitamin D levels compared to those with insufficient and deficient levels (1835 ± 252.55 vs. 1129 ± 120.7 and 1363 ± 0.125 ng/ml). Serum anti-HBsAb levels post HBV vaccination were significantly higher in women and younger individuals than in men and older individuals, respectively.  
Conclusion: This study findings suggest that participants with different serum vitamin D levels produce seroprotective antibody titers post HBV vaccination, while those with sufficient vitamin D levels may produce higher titers against HBV vaccine.
 

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Aims: The innovation ecosystem states that innovation through interactive networks occurs at different levels. The network has a wide range of stakeholders that are complex in the innovation process as part of the innovation ecosystem. Considering the importance of the issue of prevention in the health sector and the importance of the role of biotechnology in this field, the aim of this study was to examine the innovation ecosystem of human vaccines in Iran.
Participants and Methods: In this qualitative, exploratory, and descriptive research, while investigating the dimensions of the ecosystem of innovation in literature and its main characteristics, the status of the innovation ecosystem of human vaccines was investigated in Iran. This study was carried out through content analysis of the current documents and deep and semi-structured interviews with experts in this field. Subsequently, a description of the current state of the vaccine innovation ecosystem was presented.
Findings: Most of the graduates did not have enough familiarity with the techniques needed to attend the industry. The existence of two major vaccine manufacturers, the Pasteur and Razi Institutes, were of important properties of ecosystems. The small number of service providers and existing service companies with knowledge-based organizations were of shortcomings. Shortcomings in the characteristics of stable and dynamic interaction in the innovation ecosystem of human vaccines in Iran were evident and the making policies to create or strengthen these characteristics was one of the important issues of Iran in this area.
Conclusion: Despite the abundance of elements and actors in this field, the innovation ecosystem of vaccines in Iran has not yet been formulated in a structured way, and its creation and development requires the characteristics of the innovation ecosystem and the resolution of its challenges.
 

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Aims: Reports show that vaccination against influenza could elicit nonspecific immune reactions against coronavirus disease-19 (COVID-19). The present research aimed to evaluate the prevalence of COVID-19 disease among the staff of Shahid Beheshti hospital in Kashan despite vaccination against influenza.
Materials & Methods: This study was performed on 1400 employees of Shahid Beheshti hospital in Kashan from February to August 2020. Personnel whose disease was confirmed by PCR test or CT scan were considered to have COVID-19. In the present research, the relationship between influenza vaccination and the incidence of COVID-19 infection was evaluated. The collected data were analyzed by SPSS software Version 26.
Findings: Out of a total of 1400 hospital personnel participating in this study, 272 people were diagnosed as COVID-19. Among 272 patients, 23 (8.45%) cases were vaccinated. The average age of vaccinated patients was 33.48 ± 12.72 years, of whom 14 (60.87%) patients were female. Vaccination was significantly associated with prevention of COVID-19 infection (p< .05). The study of odds ratio (OR) to evaluate the effect of vaccination showed that the OR was 0.61 (95% CI: 0.39- 0.97). There was a significant difference in SpO₂, type of treatment, and lung involvement based on CT between the two groups of vaccinated and unvaccinated COVID-19 patients (p< .05).
Conclusion: In vaccinated group, COVID-19 was lower than of the no influenza vaccinated group. According to the results, the use of influenza vaccine as an effective vaccine against the new coronavirus strains could be helpful in controlling the disease.

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Objective: The global HIV epidemic continues to expand and exceeding previous predictions. An effective vaccine represents the best hope to curtail the HIV epidemic. DNA vaccines induce humoral and cellular responses and mimic live vaccines without their pathogenic potential. The importance of CD8+ CTL responses in controlling HIV and SIV viremia has led to production of a series of vaccine candidates that effectively induce these responses. It is now widely believed that an HIV vaccine strategy must stimulate both a strong humoral (antibody) as well as cell-mediated (CTL) immune response.The p24 and gp41 play many important roles in host-virus interaction and pathogenesis. These proteins are considered as attractive vaccine candidate in which their immunogenecity and immunomodulatory effects have been confirmed. Materials and Methods: In this study, a construct, pcDNA3.1Hygro- (p24-gp41), was evaluated as a DNA vaccine candidate in Balb/C mice for generation of effective cellular immune responses. For immunizing, we used dendrosome, a novel family of vehicles for transfection and therapy. IFN-γ cytokine production and total antibody were detected by ELISA. Lymphoprolifration assay was performed by MTT test. Results: ELISA and MTT assays confirmed that the cited p24-gp41 fusion gene is able to enhance immune responses in mice. Conclusion: The construct that was used in this research can be a good candidate for DNA vaccine against HIV-1, if the future complementary tests demonstrate the same trends of immunogenic responses shown in this study.

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Aims: One of the challenges of today's world and also global health priorities is pandemicity of AIDS. Studies have shown that the scope and breadth of the immune responses induction are very effective to protect against HIV. Moreover, simultaneous induction of humoral and cellular immunity responses increases the effectiveness of candidate HIV vaccines. Hence, new approaches such as polyepitopic vaccine strategy and addition of different adjuvants in HIV vaccines’ formulations have been recently considered.
Materials and Methods: In the present study, eukaryotic expression vector (pcDNA3.1-tat/pol/gag/env) was transformed and amplified in the prokaryotic host cells E. coli (DH5α). After vector extraction, it was concentrated and formulated alone and in combination with Alum adjuvant and used as DNA candidate vaccines. DNA candidate vaccines were, then, subcutaneously injected to the BALB/c mice on 0, 14, and 28 days and elicited humoral and cellular immunity responses were finally evaluated.
Findings: The results showed that the candidate DNA vaccine could not efficiently induce immunity responses (both humoral and cellular responses) by subcutaneous route injection.
Conclusion: This observation can be due to a defect in each of the steps of vector harvesting by the target cell to express the surface presentation of the epitopes on the one hand, or the inefficiency of the subcutaneous injection method on the other. Therefore, other vaccines’ injection and deliveries routes along with addition of other adjuvants in vaccine’s formulations could induce immunity responses efficiently and increase vaccine efficacy.

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Aims: COVID-19 has harmed people's lives and efforts are being made to speed up vaccinations. The growing problem of vaccine uncertainty may affect the uptake of the COVID-19 vaccine. The objectives of this study were to examine the determinants of COVID-19 vaccine acceptance.
Materials & Methods: From July 3 to September 25, 2021, we conducted a web-based, cross-sectional study among the citizens of Ardabil with a snowball sampling strategy under a highly restricted environment. A questionnaire was designed and filled out by 768 participants through social media and email. Associations between COVID-19 vaccine acceptance and determinants were explored using the chi-squared test. Key determinants that predict vaccine acceptance among respondents were modeled through logistic regression analysis.
Findings: Of the 932 survey invitees, 768 responded to the questionnaire (response rate, 82.4%). The majority (55.2%) of the study participants were female. Of the 768 respondents, 486 (63.2%) showed interest to accept the COVID-19 vaccine. Willingness to get the vaccine is relatively high among older age groups (59.4% among 40+ year old), being married (56.9%), and city dwellers participants (83.09%). In multivariate model, respondents who were above 40 years (OR: 0.7; 95% CI:0.5-0.94), and married (OR: 1.43; 95% CI: 0.97-2.09) were significantly associated with vaccine acceptance (p<0.05). Besides, people having trust in the health system and vaccine were most likely to accept the vaccine (OR: 1.26; 95% CI: 1.01-1.56), and those having a higher perceived risk of acquiring infection were 4.83 times (OR: 4.83; 95% CI: 3.78-6.17) higher odds of accepting the vaccine.
Conclusion: Our study identified religious/personal beliefs and risk perceptions as the most important predictors that would be affecting COVID-19 vaccine uptake.
 

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Aims: The success of COVID-19 vaccination depends on public acceptance of the vaccine. It is necessary to evaluate the factors affecting vaccine acceptance to increase the acceptance of vaccination. The current study aimed to determine the relationships between the three components of the COM-B (capability, motivation, and opportunity) model and the explanatory domains of each component.
Instrument & Methods: In this cross-sectional study, 1102 adults aged 18 years and older were selected through multi-stage sampling and received an online questionnaire on the WhatsApp platform in February 2021. Structure equation modeling was used to investigate the factors affecting vaccine acceptance.
Findings: Of the 1102 respondents, 938 respondents (85.1%) wanted to get vaccinated. The main indicators for the COM-B components were "behavioral regulation"(capability), "subjective norms and social support" (opportunity) and "social role" (motivation). Opportunity strongly predicted motivation (93%) and Covid-19 vaccine acceptance (74%). Motivation and capability were mediator for opportunity on vaccine acceptance.
Conclusion: Providing environmental and interpersonal conditions by creating capability and motivation in people increases vaccine acceptance.
 

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Aims: The COVID-19 pandemic has led to the global distribution of vaccines, but there are concerns regarding potential side effects. Hair loss is one of the less commonly reported side effects. The present study aimed to investigate the effect of COVID-19 vaccinations on hair loss.
Instruments & Methods: A cross-sectional descriptive study was conducted with 580 participants aged between 20 to 72 years, consisting of 270 males and 310 females. Machine learning techniques were employed to analyze the data and determine any potential relationship between COVID-19 vaccines and hair loss. A logistic regression analysis was used to assess the odds ratio and 95% confidence interval for hair loss.
Findings: Of the total participants, 17.6% reported experiencing hair loss after receiving the COVID-19 vaccine. This percentage was higher in females (19.4%) compared to the males (15.2%). There was a significant association between the COVID-19 vaccine and hair loss in both males and females. The odds ratio for developing hair loss after receiving the COVID-19 vaccine was 1.34 (95% CI: 1.04-1.73) for females and 1.12 (95% CI: 0.81-1.54) for males.
Conclusion: Hair loss is a rare but possible side effect of COVID-19 vaccination in both males and females, which its prevalence is higher in females than in males. Individuals with certain comorbidities, such as hypertension and diabetes, may be at a higher risk for experiencing hair loss after COVID-19 vaccination.
 

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In this study,the immunogenicity of streptococcosis/lactococcosis and yersiniosis vaccine in rainbow trout was investigated in farm.900 fish with an average of 50±5 g were divided into three treatments and three replications (injection treatment, immersion treatment and control group).Fish were kept for 60 days and samples were taken on the 30^th and 60^th days.Then the fishes were challenged for 14 days with three bacteria, Streptococcus iniae, Lactococcus garvieae, and Yersinia ruckeri.Sampling was done to evaluate lysozyme activity,complement, antibody titer and survival rate. The results indicated a significant increase in serum complement and lysozyme activity on the 30^th and 60^th days of sampling in the vaccinated groups compared to the control group (P<0.05).Antibody titers against S. iniae, L. garvieae and Y. ruckeri on the 30^th and 60^th days of sampling in the vaccinated groups had a significant increase compared to the control group (P < 0.05).The  relative percentage survival after 14 days of challenge with S. iniae, L. garvieae and Y. ruckeri in the injected group was (70, 60, and 76.6%),respectively, which was significant compared to the control group (P<0.05).Also, the relative percentage survival in the immersion group with S. iniae, L. garvieae and Y. ruckeri was(30, 36.6 and 53.3%),respectively, which was significant only in the group immersed with S. iniae (P<0.05).In general,it can be concluded that the use of polyvalent vaccine by injection and immersion has significant effects on the immunity and survival rate of rainbow trout. However, the injection method is more effective and suitable than the immersion method.

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Objective: Toxoplasma gondii is an obligate intracellular protozoan that causes Toxoplasmosis in human and animal. In recent years, significant progress has been made in the identification of vaccine candidates which can induce protective responses. In this study we used complete Rhoptry protein 2 gene of Toxoplasma gondii as a single DNA vaccine and evaluated its immune responses in comparison with control groups. Materials and Methods: BALB/c mice were immunized intramuscularly with three weaks time interval with pcROP2 (as case group) and pc-DNA3 and PBS (as control groups). After immunization, we evaluated the immune response using cytokine and antibody measurements. Results: The results of cytokine (IFN-γ, IL-4) assays showed that mice immunized with pcROP2, elicited stronger Th1-type cellular immune responses than those immunized with empty plasmid, or PBS (high level of IFN-γ and low-level of IL-4).Also Anti-T. gondii IgG titres (OD) increased markedly in the pcROP2 group, which was significantly higher than those of control groups (P<0.05). When challenged with the highly virulent Toxoplasma gondii RH strain, mice immunized with pcROP2 had siginificantly higher survival rates compared to control groups (P<0.05). Conclusion: This study showed that pc-ROP2 as a single DNA vaccine is effective to prime enhanced and balanced cellular and humeral immunity responses, and relatively improved mice survival time against toxoplasmosis.

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Objective: The use of bacterial plasmids carrying specific genes of pathogens as genetic vaccines is a relatively new technique for induction of cellular immune responses against microbial pathogens. Mechanisms of production of specific immune responses against these vaccines are not still completely understood. Therefore, it is necessary to examine various routes of inoculation to find the best way of immunization for specific antigens. In this research, intramuscular method of inoculation of influenza vaccine nucleoprotein (NP) encoding vector was compared with that of intra-dermal method. Materials and Methods: In this study, the ability of two different methods of immunization (intramuscular and intra-dermal) in induction of CTL responses as well as their efficiency in clearance of influenza virus from the lung of BALB/c mice was compared. Female BALB/c mice were immunized with influenza virus NP expressing plasmids on days 0, 14 and 28. CTL activity of mice was evaluated by lactate dehydrogenase technique two weeks after the last inoculation. In addition, the mice were challenged by live influenza virus and the viral titer was measured 4 days post-challenge in the lungs of animals. The results of experiments demonstrated that intramuscular immunization of mice induces a stronger CTL response. Mice immunized by intramuscular route also showed a higher ability in virus clearance from the lungs. Conclusion: Results of this study showed that different routes of immunization of influenza NP genetic vaccine induce different levels of cell-mediated immune responses and protection from the live virus.

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Objective: Several vaccines against HIV have been investigated but none has been approved as an effective HIV vaccine. An approach that could induce stronger immune response against the pathogen is utilizing a multi-epitopic vaccine. This strategy was used in the design of several vaccines and resulted in improved immune responses. Materials and Methods: In this study a multi-epitopic fusion peptide including parts of HIV-1 Nef and P24 as a vaccine candidate was injected into mice and immune humoral responses measured with total antibody and IgG sub-classes using ELISA. Also measurement of cellular immune responses through evaluation of spleen cells proliferation response using MTT and cytotoxicity by LDH were performed. Finally, the cytokine pattern of IFN-γ and IL-4 were also determined with ELISA. Results: The results indicate that candidate vaccine stimulated mouse splenic lymphocyte proliferation response and also induced strong cytotoxicity responses. Analysis of humoral immune response has shown that the candidate vaccine has induced specific antibody production mainly of the IgG2a sub-class. Also cytokine pattern evaluation has shown that IFN-γ secretion was dominant. Conclusion: The use of immunogen and conserved epitopes from P24 and Nef induced strong humoral and cellular immune responses and this construct could be candidate for further studies in animal models.

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Objective: Toxoplasmosis can lead to severe pathological effects in both infected humans and animals. The various DNA vaccines against Toxoplasma compose of single or cocktail antigens have been investigated but they have partial protective against disease. In this study, we used pcROP1 as a DNA vaccine and aluminium phosphate and aluminium hydroxide to compare their efficacy as mineral adjuvants. Materials and Methods: BALB/c mice immunized with pcROP1 alone or with co-administration of Alpo4 or Alum and the effectiveness of these two adjuvants were compared using lymphocyte proliferation assay, cytokine and antibody assay and survival time. Results: The group co-administered alum elicited stronger humoral and Th1-type cellular immune responses than the group co-administered Alpo4, while immune response in group administered with pcROP1 alone is higher than them. When challenged with Toxoplasma gondii RH strain, mice immunized with or without alum had significantly higher survival rates, whereas there was no notable enhancement of survival rate in Alpo4 group (P≤0.05). Conclusions: Our result suggest that pcROP1 plus alum and aluminium phosphate not strongly potentiate the efficacy of this DNA.

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Given the global epidemic of COVID-19, it is important to design a vaccine for prevention. The virus belongs to the beta-coronavirus family and forms appendages on the surface of the glycoprotein spike virus membrane. Studies on SARS-CoV-1 and related MERS-CoV vaccines have shown that the spike protein on the virus surface is a suitable target for the vaccine. In this experimental study, we compare the recombinant fragment of spike protein (rfsp) expressed in the eukaryotic host CHO-K1 cell and prokaryotic  E. coli in terms of immunogenicity, neutralizing activity, and epitopes recognition Similar to the virus strain and the ability to bind to the serum of improved patients, in two types of alpha and delta variants. The results showed that both rfSP proteins are a potential new antigen candidate for the development of the Covid 19 vaccine, but the CHO cell maintains the biological activity of the protein by performing post-translational modification processes such as glycolysis. This increases the present likelihood of developing virus-like epitopes and increases the titer of rfsp-specific antibodies in the serum of immunized mice. Therefore, priority is given to rfsp expressed in CHO cells to evaluate vaccine efficacy.

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Objective: The aim of this study was to construct a pcDNA3.1+ vector containing FMDV type O/IRN/1/2007-VP1 gene, protein expression in BHKT7 cells and evaluation of immune response in BALB/c mice. Materials and Methods: FMDV type O/IRN/1/2007 was isolated from a cattle in Ray in 2007 and serotyped. The purified VP1 gene was sub-cloned into the PTZ57R/T vector and pcDNA3.1+ expression vector. The PCR product of Vp1 gene without stop codon was sub-cloned upstream of EGFP gene into the pEGFP-N1 vector to evaluate VP1-GFP fusion protein expression. The pcDNA3.1-VP1 and pEGFP-VP1 vectors were transfected into BHKT7 cell line. The expression of VP1 protein was evaluated by SDS-PAGE, western blotting and florescent analysis of VP1-GFP fusion protein. The mice were injected subcutaneously by pcDNA3.1-VP1 vector as DNA vaccine and titration of neutralizing antiserum and T cell proliferation assay were done to evaluate the immune response. Results: Insertion of VP1 gene was confirmed by double digestion of sub-cloned PTZ57R/T, pcDNA3.1+ and pEGFP-N1 vectors. The specific band in western blotting was also confirmed the VP1 protein expression in BHKT7 cells. The expression of VP1-GFP fusion protein was observed under the immune-florescent inverted microscopy as more green florescent spots versus expression of GFP protein, alone. The neutralizing antiserum titer and T cell proliferation increased significantly in the group of mice vaccinated with pcDNA3.1+-VP1 vector verses control groups (P<0.05). Conclusion: The results showed that the target gene was amplified, cloned in the cloning and expression vectors and protein expression was confirmed successfully. According to the confirmed VP1 protein expression and increasing neutralizing antiserum titer and T cell proliferation by pcDNA3.1+-VP1 vector (P<0.05), it can be used as DNA vaccine against FMDV type O/IRN/2007.