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This study compares the oncolytic effect of vesicular stomatitis virus (VSV) wild type and M51R M-protein on the colorectal tumors of different invasive intensity on SW480 and HCT116 cell lines and 114 fresh colorectal cancer primary cell cultures. Fresh tumor samples were divided into two groups of lower stages (I/II) and higher stages (III/IV) regarding the medical records. The presence of two mutations in the PIK3CA gene and the expression of NEBL and AKT1 genes were evaluated. The cells were transfected with a plasmid encoding VSV wild-type and M51R mutant M-protein. Results showed either wild type or M51R mutant can kill SW480 and stage I/II primary cultures while mutant M-protein had no apoptotic effects on HCT116 cells and stage III/IV primary cultures. NEBL and AKT1 expression were significantly higher in resistant cells. Elevated caspase-9 activity confirmed that the intrinsic apoptosis pathway is the reason for cell death in lower-stage cells. Different tumors from the same cancer exhibit different treatment sensitivity due to genetic difference. NEBL and AKT1 gene expression may be responsible for this difference, which may be the target of future investigations. Therefore, tumor staging should be considered in oncolytic viral treatment as an interfering factor., Graphical Abstract, There are controversial reports about the effectiveness of a same oncolytic virus on a certain cancer type. This study explains a probable mechanism for this difference as the effect of cancer differentiation stages. NEBL and Akt1 are introduced as a molecular difference between resistant and sensitive cells to the treatment.

Background: Curcumin, a compound derived from the root of the Curcuma longa, has been confirmed as an anticancer, chemoprotective, and gene/protein regulatory agent. Nanoformulation of curcumin has been developed to increase its targeting efficiency, solubility, controlled release, and physical and chemical stability. Objectives: This study investigated the effect of new nano-type curcumin, oleic acid-derived dendrosome (OA400 nanoparticles), on the expression of E6 and E7 human papillomavirus oncogenes and P53 and Rb factors in the HeLa cell line. After preparing nano-curcumin by mixing OA400 nano-carrier and curcumin, its effect was considered on the human cervical cancer cell line (HeLa cell line RRID: CVCL_003) and normal fibroblast cells. Methods: MTT assay and flow cytometry were used to evaluate cell viability and apoptosis. Furthermore, real-time RT-PCR and western blot analyses assessed RNA and protein expression of E6, E7, P53, and Rb. Statistical analyses were performed by GraphPad Prism 7 software. Results: The nanoformulation of curcumin could reduce the expression of E6 and E7 oncogenes and increase P53 and Rb tumor suppressors in HeLa cancerous cells at 15 μM concentration; however, it had no significant effect on the viability of normal fibroblast cells. On the other hand, curcumin altered the expression of these genes at a 50-μM concentration. Gene and protein expression analysis indicated the up-regulation of P53 and Rb factors and the down-regulation of E6 and E7 under the influence of nano-curcumin treatment more than curcumin. Conclusions: These data indicate the potential of curcumin-loaded OA400 nanoparticles to be considered as a treatment option in cervical cancer investigations.

Coronavirus disease 2019 (COVID-19) is an infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), but the pathogenesis is unclear. Host genetic background is one of the main factors influencing the patients' susceptibility to several viral infectious diseases. This study aimed to investigate the association between host genetic polymorphisms of two genes, including vitamin D receptor (VDR) and vitamin D binding protein (DBP), and susceptibility to COVID-19 in a sample of the Iranian population. This case-control study enrolled 188 hospitalized COVID-19 patients as the case group and 218 suspected COVID-19 patients with mild signs as the control group. The VDR (rs7975232, rs731236 and rs2228570) and DBP (rs7041) gene single nucleotide polymorphisms (SNPs) were genotyped by Polymerase Chain Reaction Restriction - Fragment Length Polymorphism (PCR-RFLP) method. A significant association between rs2228570 SNP in the VDR gene and the susceptibility of COVID-19 was found between case and control groups. The CT genotype (Heterozygous) of rs2228570 C T polymorphism showed significant association with a 3.088 fold increased odds of COVID-19 (p .0001; adjusted OR: 3.088; 95% CI: 1.902-5.012). In addition, a significant association between CC genotype of rs2228570 CT polymorphism and increased odds of COVID-19 in male and female groups (p = .001; adjusted OR: 3.125; 95% CI: 1.630-5.991 and p = .002; adjusted OR: 3.071; 95% CI: 1.485-6.354 respectively) were determined. Our results revealed no significant differences in the frequency of genotype and allele of VDR (rs7975232 and rs731236) and DBP (rs7041) between SARS-CoV-2-infected patients and controls (p .05). Our results showed that polymorphism of VDR (rs2228570) probably could influence individual susceptibility to COVID-19. The polymorphisms of VDR (rs7975232 and rs731236) and DBP (rs7041) were not associated with SARS-CoV-2 infection susceptibility.

Human cosavirus (HCosV) is a newly recognized virus that seems to be partly related to nonpolio flaccid paralysis and acute gastroenteritis in pediatric patients. However, the relationship between HCosV and diseases in humans is unclear. To assess an investigation for the occurrence of HCosV among pediatric patients involved in meningitis and encephalitis, we implemented a real-time quantitative polymerase chain reaction assay for detection and quantification of HCosV in stool specimens.In this study, a total of 160 cerebrospinal fluid samples from September 2019 to October 2020 were collected from presenting pediatric patients with meningitis and encephalitis in a Karaj hospital, Iran. After viral RNA extraction, the real-time quantitative polymerase chain reaction was performed to amplify the 5'Un-Translated Region region of the HCosV genome and viral load was analyzed.Of the 160 samples tested, the HCosV genomic RNA was detected in 2/160 (1.25%) of samples. The minimum viral load of HCosV was 3.5 × 103 copies/mL from 4 years male patient. The maximum viral load was determined to be 2.4 × 105 copies/mL in one sample obtained from 3.5 years female patient.This is the first documentation of HCosV detection in cerebrospinal fluid samples that better demonstrates relation of HCosV with neurologic diseases including meningitis and encephalitis. Also, these results indicate that HCosV has been circulating among Iranian pediatric patients.

Background: Oncolytic virotherapy is a promising therapeutic approach for several cancers which however has been reported that is ineffective in some cases of the same cancer. A well-known oncolytic virus is the Vesicular stomatitis virus (VSV) due to activating the apoptosis mechanism in tumor cells through its Matrix (M) protein. This study compares the different invasive intensity Colorectal tumors for the oncolytic effect of VSV wild type (wt) and M51R M-protein.Methods: 114 fresh colorectal tumor primary cell cultures plus SW480 and HCT116 colorectal cancer cell lines were examined. Fresh tumor samples were divided into two groups of lower stages (I/II) and higher stages (III/IV) regarding the medical records. The presence of two mutations in the PIK3CA gene, and the expression of NEBL and AKT1 genes were evaluated to biologically compare the staging classification. The cell lines and the stable primary tissue cultures were transfected with a plasmid encoding VSV wild type and M51R mutant M-protein. Western blotting assay confirmed the expression of the protein. Results: MTT assay results showed either wild type or M51R mutant can kill SW480 and stage I/II primary cultures while mutant M-protein had no apoptotic effects on HCT116 cells and stage III/IV primary cultures. Morphological apoptosis features were observed by a phase-contrast microscope. NEBL and AKT1 expression were significantly higher in resistant cells. Elevated Caspase 9 activity confirmed that the intrinsic apoptosis pathway is the reason for cell death in SW480 cells. The apoptosis rate was quantified and reconfirmed by Annexin V FITC/PI multicolor flow-cytometry. Conclusions: Different tumors from the same cancer exhibit different sensitivity to M51R M-proteins due to genetic difference. NEBL and AKT1 gene expression may be responsible for this difference which maybe the target of future investigations. Therefore, tumor staging should be considered in oncolytic viral treatment as an interfering factor.

Cervical cancer is a common cancer in women around the world. Human papillomavirus infection is main cause of this cancer. Currently treatment methods include chemotherapy, radiation therapy and surgery. These methods usually cause severe reactions and drug resistance in the patients, so finding a safe natural drug for treatment can be very helpful. Curcumin is a compound derived from the root of the Curcuma longa plant and is the active ingredient in a food spice. Various studies have confirmed the anti-cancer, chemoprotective and genes and proteins regulatory effects of this component. Insolubility, low cellular uptake and low bioavailability are common limitations of curcumin use. To overcome these limitations, a nano-formulation of curcumin has been developed in our laboratory. In this study, the effect of nanocurcumin on cellular apoptosis induction of Hela cancer cells was compared with curcumin. It has demonstrated that nanocurcumin function is three time more effective than curcumin in same concentration. This nano-formulation of curcumin in 15µM concentration was able to reduce the expression of E6 and E7 oncogenes, and increase P53 and Rb tumor suppressors in HeLa cancerous cells with p-value = 0.001 − 0.0001, whereas curcumin altered these genes expression in 50µM concentration with p-value = 0.05 − 0.01. Nanocurcumin has no significant effect on the viability of normal fibroblast cells. Therefore, this compound has no effect on normal cells. These findings are consistent with the results of previous studies about nanocurcurcumin in other types of cancer. Therefore, nanocurcumin may be a suitable option to consider for cervical cancer treatment.

Cervical carcinoma is the third leading cancer in women. There is a possibility of treatment failure of conventional chemotherapy due to the resistance of cancer cells to the apoptotic effects of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). The use of herbal drugs for the treatment of cancer has been considered an effective solution. Chelidonium majus L. (C. majus) alkaloid extract can induce apoptosis on cancer cells. This study examined how the crude alkaloid extract of C. majus on HeLa cells is mediated apoptosis by evaluating DR4, DR5, and FADD as death receptors. The cytotoxicity of Chelidonium majus L. alkaloid extract on HeLa and human fibroblast cells was assessed using an MTT assay. The percentage of apoptotic cells was evaluated by flow cytometry. The gene expression levels of DR4, DR5, FADD, caspase 8, and caspase 3 were evaluated by real-time reverse transcription-polymerase chain reaction (RT-PCR) assay. The half-maximal inhibitory concentration (IC50) of Chelidonium majus L. alkaloid extract was determined 75 μg/ml and 860 μg/ml for HeLa and fibroblast cells, respectively, after 48 h. The subtoxic concentration of extract with 65% viability was determined 50 μg/ml for the HeLa cell line. Quantitative RT-PCR analysis demonstrated a significant increase in DR4, DR5, caspase 8, and caspase 3 mRNA expression at both 50 μg/ml and 75 μg/ml concentrations of extract compared to control. In contrast, FADD gene expression did not significantly differ at any of 50 μg/ml and 75 μg/ml concentrations. It was demonstrated in this study crude alkaloid extract of C. majus induces an extrinsic apoptotic pathway through DR4, DR5 in the HeLa cell line and can be a natural product candidate for exerting synergism for cervical carcinoma chemotherapy. Further investigations are needed to clarify the essential components of C. majus alkaloid extract that induces apoptosis.

The human T-lymphotropic virus type 1 (HTLV-1) can cause ATL or TSP. This study evaluates the prevalence of HTLV-1 infection in blood donors in Golestan province. The study was conducted among 4226 blood donors and ELISA test was performed for the initial HTLV-1 screening. Reactive samples were confirmed by Western blot and Electrochemiluminescence tests. Then recalling donors with reactive results was done and genomic DNA from the new sample was extracted and tested using the Nested PCR method and phylogenetic analysis was performed. At first, 8 samples were reactive with ELISA test and 4 samples were confirmed with western blot, Electrochemiluminescence and Nested PCR tests.The sequences of isolates was related to the HTLV-1 virus and subtype a (cosmopolitan) subgroup A.The prevalence of HTLV-1 virus in Golestan province was about 0.09 %.The genotype of virus isolates had a common ancestor with isolates of the Khorasan region.

In the present study, we developed a novel approach for functionalization of gelatin nanofibers using the plasma method for tissue engineering applications. For this purpose, tannic acid-crosslinked gelatin nanofibers were fabricated with electrospinning, followed by treatment with argon and argon&ndash, oxygen plasmas in a vacuum chamber. Samples were evaluated by using scanning electron microscopy (SEM), atomic force microscopy (AFM), attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, contact angle (CA) and X-ray diffraction (XRD). The biological activity of plasma treated gelatin nanofibers were further investigated by using fibroblasts as cell models. SEM studies showed that the average diameter and the surface morphology of nanofibers did not change after plasma treatment. However, the mean surface roughness (RMS) of samples were increased due to plasma activation. ATR-FTIR spectroscopy demonstrated several new bands on plasma treated fibers related to the plasma ionization of nanofibers. The CA test results stated that the surface of nanofibers became completely hydrophilic after argon&ndash, oxygen plasma treatment. Finally, increasing the polarity of crosslinked gelatin after plasma treatment resulted in an increase of the number of fibroblast cells. Overall, results expressed that our developed method could open new insights into the application of the plasma process for functionalization of biomedical scaffolds. Moreover, the cooperative interplay between gelatin biomaterials and argon/argon&ndash, oxygen plasmas discovered a key composition showing promising biocompatibility towards biological cells. Therefore, we strongly recommend plasma surface modification of nanofiber scaffolds as a pretreatment process for tissue engineering applications.

Background Host genetic factors and single nucleotide polymorphism (SNP) in host genes play important roles in the progression of disease stages and response to treatment of viral infections, including hepatitis B infection. Vitamin D plays a key role in a wide range of biological activities such as the immune response, proliferation and cellular differentiation. Since the biological effects of this vitamin are affected by the Vitamin D receptor (VDR) and the vitamin D-Binding Protein (DBP), in this study we investigated the association between VDR and DBP SNPs with the risk of chronic hepatitis B (CHB) infection in Iranian patients. Methods Our case-control study consisted of two groups including 100 patients with CHB and 100 healthy controls. SNP of ApaI (rs7975232), TaqI (rs731236), BsmI (rs1544410) and FokI (rs2228570) VDR-related and DBP (rs7041) gene loci were determined by PCR-RFLP method. Serum ALT and AST levels and HBV virus load were determined and its association with SNP was investigated. Statistical analysis was performed using SPSS software (V22.0). Results The results of this study showed that the TT (Homozygous Mutant) genotype of FokI locus (C>T; rs2228570) compared with CC (Homozygous Wild Type) and CT (Heterozygous) genotypes may be associated with an increased risk of CHB infection (P; 0.012, OR 2.674 (1.233–5.800), 95% CI). The T allele was also significantly associated with CHB infection (P; 0.009, OR 1.687 (1.136–2.506), 95% CI) compared to the C allele FokI. In this study, no significant relationship was found between CHB infection and rs7975232, rs731236, rs1544410 and rs7041 polymorphisms (P > 0.05). Also, there was no significant relationship between serum ALT and AST levels and HBV Viral load in patients with rs2228570 polymorphism (P > 0.05). Discussion According to the results, VDR FokI (rs2228570) polymorphism has an impact on the chance of chronicity of hepatitis B in Iranian patients.

Herpes simplex virus type 1 (HSV-1) causes serious infections particularly in immunocompromised patients. Methanolic extract of four plants were evaluated for their anti-viral effects against acyclovir resistant HSV-1 in HeLa cell line. The 50% cytotoxic concentration (CC50) as well as the effective minimal cytotoxic concentration of each plant extract were evaluated by MTT assay. Antiviral effects of the plant extracts on HSV-1 were examined at different concentrations of the extract. The effective minimal cytotoxic concentration was evaluated at different times of virus replication after infection. Virus titration was assessed by tissue culture infectious dose 50 (TCID50) method. Among the 4 plant extracts evaluated only Mentha pulegium L. extract was shown to exert the highest antiviral activity, with selectivity index (SI) 10.25. Direct treatment of HSV-1 with Mentha pulegium L. extract resulted in 1.7 log10 TCID50 reduction in virus titers after one hour. The highest reduction in HSV-1 infectivity was obtained 1 hour after the infection of the cells with virus resulting in 2.1 log10 TCID50 reduction as compared to the control. The antiviral effects of Mentha pulegium L. extract on HSV-1 after virus infection was more remarkable than the virucidal activity.

Background Acute respiratory infection plays an important role in hospitalization of children in developing countries; detection of viral causes in such infections is very important. The respiratory syncytial virus (RSV) is the most common etiological agent of viral lower respiratory tract infection in children, and human metapneumovirus (hMPV) is associated with both upper and lower respiratory tract infections among infants and children. Objectives This study evaluated the frequency and seasonal prevalence of hMPV and RSV in hospitalized children under the age of five, who were admitted to Aliasghar children’s hospital of Iran University of Medical Sciences from March 2010 until March 2013. Patients and Methods Nasopharyngeal or throat swabs from 158 hospitalized children with fever and respiratory distress were evaluated for RSV and hMPV RNA by the real-time polymerase chain reaction (PCR) method. Results Among the 158 children evaluated in this study, 49 individuals (31.1%) had RSV infection while nine individuals (5.7%) had hMPV infection. Five (55.5%) of the hMPV-infected children were male while four (44.5%) were female and 27 (55.2%) of the RSV-infected patients were females and 22 (44.8%) were males. The RSV infections were detected in mainly < one year old children and hMPV infections were detected mainly in > one year old children. Both RSV and hMPV infections had occurred mainly during winter and spring seasons. Conclusions Respiratory syncytial virus was the major cause of acute respiratory infection in children under one-year of age while human metapneumovirus had a low prevalence in this group. The seasonal occurrence of both viruses was the same.

Although CD8 + cytotoxic T lymphocyte (CTL) epitope-based DNA vaccination is valuable experience on vaccine research but many attempts are still continued to achieve acceptable protective response. To study the role of full length antigen in CTL epitope immunization, we evaluated cellular immunity of diverse patterns of complete Herpes simplex virus type 1 ( HSV-1 ) glycoprotein B (gB) and the immunodominant CTL epitope (498–505) DNA injection in C57BL/6 mice. Optimal immune response was observed in the group immunized with the full length of gB in the first injection and CTL epitope in the second and third vaccination as assessed by lymphocyte proliferation assay (MTT), cytokine assay (ELISA) and CTL assay. B cell and spatially CD4 + T cell epitopes in full length protein might be important for appropriate priming of CTL immune response. These findings may have important implication for the improvement of CTL epitope based DNA vaccine against HSV and other pathogens.

The immunomodulatory effects of exogenous opioids on induction of acquired immunity during microbial infection are now well known; however, our knowledge about the relationship between endogenous opioid response and microbial infections is rudimentary. Here, we report the effect of administration of Naloxone (NLX), an opioid receptor antagonist, on induction of acquired immunity during primary herpes simplex virus type 1 (HSV-1) infection. BALB/c mice received NLX, twice daily, 2 h before infection with HSV-1 until 7 days after infection. Cell-mediated immunity was assessed by evaluating lymphocyte proliferation, interferon- γ (IFN- γ ) production, delayed type hypersensitivity (DTH) and mortality rate after acute HSV-1 challenge. The findings showed that a higher level of cell-mediated immunity was induced in the NLX-treated animals compared to the control group after induction of HSV-1 infection. However, the data indicate similar neutralizing antibody production in NLX-treated animals and control animals. This observation and further studies in this field may lead to the use of NLX as an adjuvant for designing microbial vaccines and adjunctive therapy of viral infections.

Gamma interferon (IFN- γ ) is among the most important immune factors for limiting of herpes simplex virus (HSV) infections. However, our knowledge about the kinetics of IFN- γ production after HSV infection is limited. The present study examines the kinetics of IFN- γ expression following secondary infection with HSV-1. Using semiquantitative RT-PCR assay, the expression of IFN- γ in spleen lymphocytes was significantly detected on 14 days but not 7 days after intraperitoneal inoculation of HSV-1, while ELISA detected IFN- γ on both days. At various hours after in vitro re-stimulation of spleen cells, RT-PCR showed a decreasing pattern of mRNA transcripts, whereas, ELISA assayed an increasing amount of secreted protein through the experiment. Despite the contrast results of ELISA and RT-PCR, regarding the short half-life of mRNA, the data are in correlation with each other and need to interpret.

Improving vaccine potency in the induction of a strong cell-mediated cytotoxicity can enhance the efficacy of vaccines. Necrotic cells and the supernatant of necrotic tumor cells are attractive adjuvants, on account of their ability to recruit antigen-presenting cells to the site of antigen synthesis as well as its ability to stimulate the maturation of dendritic cells.To evaluate the utility of supernatant of necrotic tumor cells as a DNA vaccine adjuvant in a murine model.The supernatant of EL4 necrotic cells was co-administered with a DNA vaccine expressing the glycoprotein B of Herpes simplex virus-1 as an antigen model under the control of Cytomegalovirus promoter. C57BL/6 mice were vaccinated three times at two weeks intervals with glycoprotein B DNA vaccine and supernatant of necrotic EL4 cells. Five days after the last immunization, cell cytotoxicity, IFN-γ and IL-4 were evaluated.The obtained data showed that the production of IFN-γ from the splenocytes after antigenic stimulation in the presence of the supernatant of necrotic EL4 cells was significantly higher than the other groups (p0.002). The flow cytometry results showed a significant increase in the apoptosis/necrosis of EL4 cells in the mice immunized with DNA vaccine and supernatant of necrotic EL4 cells comparing to the other groups (p0.001).The supernatant of necrotic cells contains adjuvant properties that can be considered as a candidate for tumor vaccination.

Studies on efficacy of various vaccines that prevent or reduce the primary and recurrent HSV-1 infection have demonstrated the importance of cellular immunity for protection against the infection. We previously used DNA vaccination to induce cellular immunity against HSV-1 infection in mice.The aim of our study was to evaluate the effect of LIGHT; a member of TNF super family, on the kinetic of CTL response induced by HSV-1 glycoprotein B based DNA vaccine.Using a granzyme B ELISA for detection and analysis of CD8+ T cells, CTL activity was determined in the spleen of BALB/c mice at various time points after primary and booster dose of vaccination. The kinetics of CTL response to primary and secondary HSV-1 infection and DNA vaccination were compared to those induced by DNA vaccination in combination with LIGHT adjuvant in the present study.In primary and secondary immunization, the CTL activity in the HSV injected group peaked 7 days and 12 hours post immunization, respectively. After 5 days, LIGHT could neither accelerate the CTL response compared to DNA vaccination alone nor could enhance the CTL activity in the primary and the first peak of memory response, the amount of granzyme B induced by the LIGHT containing vaccine was significantly higher than that induced by the vaccine without the adjuvant.Although LIGHT enhances the cellular response in the booster dose of vaccination, it does not accelerate the CTL response.

Many approaches have so far been tried to enhance the immunogenicity of DNA vaccine. These include the use of various factors that induce apoptosis or anti-apoptosis effects when co-delivered with DNA vaccine. In the present study, the effects of pro-apoptotic Bax encoding plasmid (pBax) and anti-apoptotic Bcl-X(L) encoding plasmid (pBcl-xl), intradermally co-injected with glycoprotein B (gB) of Herpes Simplex Virus (HSV)-1 encoding plasmid (pgB) into the C57BL/6 mice were evaluated. Immune responses of the mice to the antigen were assessed by antibody assay, lymphoproliferative responses as well as cytokine and cytotoxic T-lymphocyte (CTL) assay. Analysis of the humoral and cellular responses showed that the mice immunized with pBax and pgB induced higher levels of antibody and Interleukin-4 as well as stronger lymphocyte proliferative responses and cytotoxic activity compared to those mice received pgB alone. pBcl-xl when intradermally co-injected with pgB showed no significant enhancement in immune responses comparing to pgB.

Although the role of various cytokines on stimulating the immune responses is characterized well, the importance of LIGHT, a member of TNF superfamily, is less clear. In the current study, we administrated LIGHT expression plasmid as an adjuvant to HSV-1 gB DNA vaccine. HSV-1 gB DNA can elicit vigorous humoral and cell mediated immunity in BALB/c mice. LIGHT could potentiate the proliferation of T lymphocytes and induction of T CD8+ cells performing by measuring Granzyme B, a specific marker of CMI immunity and virus neutralization antibody titer. In this study, timing effect of cytokine administration on the resultant immune pattern was evaluated in three different timing groups. The group received LIGHT 3 days before DNA vaccine, demonstrated significant increase in cell mediated immunity. So, utilization of an adjuvant to DNA vaccine can significantly influences the induced immune response consequently and this phenomenon could be important to obtain the optimal response in DNA vaccine strategy. Given the growing use of plasmid-based immune adjuvants to improve the immunogenicity and efficacy of DNA vaccines, these findings support the need for further detailed study of this class of agent.

Herpes simplex virus type 1 is one of the most common viruses among human population. Studies demonstrate the essential role of cell mediated immunity, especially CD8+ T cells, in prevention and clearance of HSV1.It is of great importance to improve our knowledge about the kinetics of CTL responses to primary and secondary HSV-1 infection.Using a sensitive technique for detection and analysis of CD8+ T cells, granzyme B ELISA, the CTL activity in the spleens of Balb/c mice at various time points after intraperitoneal administration of HSV1 (strain KOS) in primary and secondary infections were determined.During acute HSV-1 infection, virus specific cytotoxic T cells were detected at day 5 post virus inoculation and peaked at day 7. Six hours after secondary infection the activity of memory CD8+ T cells was detected and peaked at 12 hours post infection.The peak of CTL activity was found to be day 7 post infection in primary HSV-1 infections which decreased with time. In secondary infections, the activity of CTLs reached the highest level at 12 hours post infection.

The vast majority of the world's population is infected with Herpes simplex virus (HSV). Although antiviral therapy can reduce the incidence of reactivation and asymptomatic viral shedding, and limit morbidity and mortality from active disease, it cannot cure infection. Therefore, the development of an effective vaccine is an important global health priority. In this study, the induction of IFN-gamma production was compared by different herpes simplex virus 1 (HSV1) vaccines. Glycoprotein D (gD1) as a major immunogenic HSV1 glycoprotein was chosen to our study. Balb/c mice were administered with DNA vaccine encoding gD1, subunit glycoprotein vaccine including insect cells infected by a gD1 recombinant Baculovirus, prime DNA vaccine boosted by subunit glycoprotein vaccine, inactivated KOS strain as a positive control, PcDNA3 plasmid and Sf9 cells as a negative control. Evaluation tests showed kinetics of IFN-gamma mRNA at 8, 16 and 32 hours after restimulation sharply decreased whereas, IFN-gamma protein is significantly increased. Our results revealed that at 14 days after immunization IFN-gamma secretion of stimulated cells in all of the vaccinate groups dramatically raised rather than secreted IFN-gamma levels in mice that were analyzed at 7 days after vaccination. In comparison to other groups; Prime-Boost immunization dramatically caused vigorous and prompt IFN-gamma production at 7 days after immunization and 8 hours after restimulation.