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According to some difficulties against tuberculosis (TB) vaccination, development of new TB vaccines has been noted in recent years. Selection of proper route for vaccination is one of the most important factors for induction of good immune responses. Hence, in this study, the effects of different administration routes, including intranasal (I.N), subcutaneous (S.C) and intramuscular (I.M) on immune responses against Mycobacterium tuberculosis ESAT-6/CFP-10 recombinant protein has been considered. Recombinant ESAT-6/CFP-10 protein with or without adjuvant (MF59 or cholera toxin B (CTB)) was administered by three routes of I.M, I.N and S.C to mice for three times. Then, the levels of specific antibodies, lymphocyte proliferation and IFN-γ/IL-5 cytokine profile have been carried out to evaluate the humoral and cellular responses. The results showed that the titers of specific antibodies were quickly elevated in S.C and I.M groups after first immunization. Otherwise, the raise of antibody has delay in the I.N immunized animals. The levels of IFN-γ and lymphocyte proliferation have been increased in all of vaccinated groups. However, the I.N immunized mice have lower levels of IL-5 production. Based on our finding, the ESAT-6/CFP-10 recombinant protein is a potent stimulator of immune responses in all of three immunization strategies. However intranasal administration of this antigen has tended to reinforcement of cellular immune responses. [ABSTRACT FROM AUTHOR]

Objective: Reduction of cerebral ischemia-reperfusion injury (IRI)/re-oxygenation injury, is defined as the paradoxical exacerbation of the cellular dysfunction and death, following restoration of the blood flow to previously ischemic tissues. The re-establishment of blood flow is essential to salvage the ischemic tissues. As a result, the treatment of IRI with novel therapies, which have fewer side effects, are of great importance. Therefore, this study aimed to investigate the effects of curcumin nanoparticle (CN) pre-treatment on the cerebral I/R rat model. Materials and Methods: In this experimental study, CN was administered to rats orally five days before the bilateral common carotid artery occlusion (BCCAO) and continued for three days. The intensity of oxidative stress, the activities of antioxidant enzymes, glutathione (GSH) content, the activity of mitochondrial enzymes, including succinate dehydrogenase (SDH), malate dehydrogenase (MDH) and lactate dehydrogenase (LDH), curcumin bioavailability, pERK/ERK expression ratio and TFEB protein were studied. Data analysis was performed using Graphpad Prism V.8 software, one-way analysis of variance (ANOVA) with the statistical package for the social sciences (SPSS V.26 software). Results: Cerebral IRI-damage significantly increased the oxidative stress (P=0.0008) and decreased the activity of the antioxidant enzymes including catalase (CAT) (P<0.001), super oxide dismutase (SOD) (P<0.001), reduced GSH (P<0.001), mitochondrial enzymes, pERK/ERK expression ratio (P=0.002) and TEFB protein (P=0.005) in rats’ brains. In addition, the pre-treatment of the rats with CN resulted in a decrease in the reactive oxygen species (ROS), and an increase in the activities of antioxidants and mitochondrial enzymes. This in turn up-regulated the pERK/ERK expression ratio and TEFB expression. Conclusion: CN has neuroprotective effects on the cerebral IRI condition due to its antioxidant properties and is able to overexpress the pERK and TFEB proteins; thus, it can be considered as a suitable treatment option during and after the incidence of stroke. [ABSTRACT FROM AUTHOR]

The aim of this study was to design a multiepitope universal vaccine for major human papillomavirus (HPV) structural proteins, L1/L2, by bioinformatics models. For this purpose, we predicted the most probable immunogenic epitopes of L1 and L2 from common high-risk HPV 16, 18, 31, and 45 beside high prevalent type 6 and 11 based on BCPREDS defaulted model, while solvent accessibility of structure was extrapolated. The three-dimensional molecular model of L1 protein was constructed by Swiss Model server, whereas sequence alignment provided model for prediction of L2 protein epitopes. After that, N-glycosylation sites were excluded from estimated epitope regions. Then, by other bioinformatics analyses, 20 epitopes were selected and fused in tandem repeats, reverse translated, and codon optimized to relevant sequence. The final protein parameters such as antigenicity were analyzed by protean program. Evaluation of new recombinant protein sequence indicated a molecular weight of 41.8 kDa with 400 amino acids beside positive charge. The computed isoelectric point (pI) value indicated the acidic nature of final product. The aliphatic index showed low thermal stability of this construct and the Grand Average Hydropathicity value was negative (−0.494). Analyzed plot showed that major parts of new protein construct had hydrophilic property, thus harboring antigenic potency. After all, sequence of final construct reverse translated to DNA and this codon-optimized sequence showed Codon Adaptation Index (CAI) of >0.8 for expression in Escherichia coli. Finally, this sequence ligated into pET28a bacterial expression vector. The new recombinant proteins harboring 20 B cell epitope seem to be suitable antigens based on computational methods as a universal vaccine candidate for HPVs. [ABSTRACT FROM AUTHOR]

IntroductionThe early secreted antigenic target 6-kDa protein (ESAT-6) plays an important role in immune protection against Tuberculosis. Owing to its great potential to increase immune response, chitosan can be considered as a suitable biodegradable polymer for intranasal administration.MethodsThe physiochemical properties of the nanoparticle were measuredin vitro. Two weeks after the last intranasal administration, blood samples were collected and specific IgG, IFN-gama, and IL-4 levels were measured by ELISA.ResultsChitosan nanoparticles containing ESAT-6 demonstrated stronger ability to induce IFN-gama, IL-4, and IgG antibody level than the control groups. Conclusion Administration of chitosan nanoparticles can be a suitable method to induce more appropriate immune responses against low inherent immunogenic tuberculosis proteins through intranasal routs. [ABSTRACT FROM AUTHOR]

Abstract: Tuberculosis (TB) remains as a major public health problem worldwide. Identification and selection of immunodominant antigens of Mycobacterium tuberculosis (MTB), capable of efficiently inducing a protective immune response is the ultimate goal of TB vaccine development studies. Accordingly, this study was designed to produce a novel M. tuberculosis fusion protein consisted of MTB ESAT-6 (early secreted antigenic target-6 kDa), as a potent immunogenic protein, fused to C-terminus of MTB HSP70 (HSP70359–610), as an appropriate carrier and adjuvant. The constructed gene was inserted into a prokaryotic expression vector (pQE30); consequently, the recombinant fusion protein with a 6xHis-tag was successfully over expressed in Escherichia coli M15. Inclusion bodies from bacterial cell lysates were solubilized and the recombinant fusion protein was easily purified by Ni-NTA affinity chromatography under denaturing conditions followed by urea gradient dialysis. The purified and refolded protein was then applied for immunization of mice that resulted in the detection of high titers of specific antibodies, high level of IFN-γ and cell proliferation. The results of our study could confirm the capability of E6H70C fusion protein, as a potential tuberculosis vaccine candidate, for the efficient induction of specific immune responses in a mouse model. However, further investigation need to evaluate the protectivity of this recombinant protein in host model. [Copyright &y& Elsevier]

Abstract: Due to its conservation, the extracellular domain of the influenza A M2 protein (M2e) has the potential for being applied as a recombinant vaccine candidate against a wide range of strains, though its immunogenicity may need to be improved. The occurrence of several post-translational modifications within the structure of M2 protein may affect its immunopotency for the induction of humoral immune response. Herein, to construct a recombinant M2e-based vaccine candidate with the appropriate structural conformation and immunogenicity the corresponding nucleotide sequence from an H9N2 influenza strain was fused to the N-terminus of the truncated Mycobacterium tuberculosis HSP70359–610, as a potent adjuvant, and following its cloning into the pPICZαA plasmid the fusion gene was expressed in Pichia pastoris KM71H yeast. The secreted protein was then easily purified from the culture media, based on the presence of polyhistidine tag and used for the production of rabbit polyclonal antisera. This raised antisera could recognize the native M2e protein on the surface of H9N2 influenza virus-infected MDCK cells at a comparable level with the commercial H2N2-specific anti-M2 antibody, which was evidenced with immunofluorescence and cell–ELISA assays. These results not only re-emphasized on the conservancy of the M2e antigen, but also pointed towards the applicability of the M2e–HSP70359–610 fusion protein for the induction of specific antibodies capable of binding to the native M2e antigen on the infected cells. Collectively, this study implied that purified M2e–HSP70359–610 represents a promising vaccine candidate; however, its in vivo potency for the induction of protection remains to be evaluated. [Copyright &y& Elsevier]

Objective(s): According to the occupationally risk of infection in staff workers who have direct contact with mycobacterium species, we investigated their immunological parameters and compared with healthy purified protein derivative (PPD) negative volunteers. Materials and Methods: We investigated 20 PPD positive volunteers working at Tuberculin Unit of Razi Vaccine and Serum Research Institute and PPD negative healthy controls with no exposure or history of active tuberculosis. The percentages of circulating lymphocyte subpopulations were detected by flowcytometry. IL-4 and IFN-γ production levels were measured by ELISA in supernatants of PPD-stimulated peripheral blood mononuclear cells (PBMCs) culture. Results: Tuberculin workers showed an increase in IFN-γ level and significant decrease of CD4+ T cells percentage and CD4/CD8 ratio compared to PPD negative normal individuals. However the IL-4 production and percentage of other lymphocyte population has been unchanged. Discussion: These observations suggest that the immunological parameters of tuberculin workers with PPD positive reaction, who are occupationally exposed to mycobacterium antigens, could be changed. Future studies will be directed towards cytokine networking and regulatory lymphocytes, which will help us validate the significant data presented in this study. [ABSTRACT FROM AUTHOR]

Introduction: The virulent Newcastle disease virus (NDV) strains cause an economically important infectious disease in poultry. The common vaccination program with genotype II NDV strains is routinely practiced to provide a better protection level against Newcastle disease (ND). Nevertheless, the emergence of new antigenic and genetic variants within the circulating NDVs raises the importance of improved control strategies. The genotype VII NDV is associated with many of the most recent outbreaks of the disease worldwide. Methods: We evaluated the impacts of genetic divergence between the genotypes II and VII on the immunity against NDV to choose a suitable vaccine virus candidate by focusing on the F and HN proteins. Comparative bioinformatics analyses based on B- and T-cell epitopes binding affinity, protein secondary structure and physicochemical properties predictions were applied for genotypes II and VII. Results: Although the results showed more differences in HN protein than F protein, there was no major difference between the predicted antigenicity values, epitope regions, affinity binding to MHC-I and MHC-II, secondary structures, surface accessibility, and stability of these immunogens between genotypes II and VII. Conclusion: The results suggest that genotype II-based live vaccines can induce immune responses against NDV; however, an inactivated vaccine formulated by genotype VII should be considered in combination with the traditional live vaccine to provide better protection in controlling programs against ND. [ABSTRACT FROM AUTHOR]

Asthma is an inflammatory disease of the respiratory system affecting over 300 million people worldwide. Lavandula has been traditionally used to treat inflammatory diseases and has also several medical applications. We aimed to explore the anti-inflammatory effects of Lavandula extract on a murine model of asthma. Twenty-eight Balb/c mice were divided into four groups of seven animals. The negative control group was treated with Phosphate-buffered saline, and the remaining three groups received ovalbumin to induce asthma. Subsequently, two asthmatic groups received budesonide or Lavandula extract. Afterward, the changes in the number of eosinophils, gene expressions, and the levels of interleukins 5, 13, and 33 were quantified in bronchoalveolar lavage fluid (BALF) of animals. Histopathological alterations of goblet cells, mucus secretion, and peribronchial and perivascular inflammations were examined in lung tissues of all animals. Eosinophils count and the levels of interleukins 5, 13, and 33 in BALF specimens were significantly reduced. Hyperplasia of goblet cells, mucus hypersecretion, and peribronchial and perivascular inflammations were inhibited in mice treated with Lavandula compared to asthmatic group. Interestingly, Lavandula was more effective than budesonide in alleviating inflammatory responses of asthma. The present study is the first report of anti-inflammatory and antiasthmatic activities of Lavandula extract in the Balb/c mouse model of ovalbumin-induced asthma. Lavandula antiasthmatic effects were mediated by modulating eosinophil counts, type 2 inflammatory responses, mucus secretion, and histological alterations of the lung due to inflammation, proposing it as a therapeutic modality to attenuate inflammatory complications of asthma. [ABSTRACT FROM AUTHOR]

Background and Objectives: One of the highly conserved outer membrane proteins expressed only by pathogenic Leptospires is Loa22. The study aims is to achieve the optimum conditions for high expression of recombinant Loa22 (rLoa22) protein. Materials and Methods: Complete coding sequence of loa22 gene sub-cloned into a pET32a (+) expression vector. BL21 competent E. coli (pLysS) used as expression host for transformation. The recombinant clones selected on ampicillin plates and subjected to PCR by using pET T7 primers. Then expression conditions optimized by adjusting parameters such as culture media, induction time, temperature, and IPTG concentration. Results: SDS-PAGE analysis showed that high production of rLoa22 protein obtained when post induction incubation, IPTG concentration, and duration of induction were 37°C, 0.1 M and 5 h in 2×TY medium respectively. The purification of rLoa22 protein under native conditions using Ni-NTA pull-down was optimum in one hour binding at 37°C, five times washing process and elution buffer with a pH 7.4 and a 0.3 M imidazole concentration. Conclusion: The findings of the study led to high production of pure Loa22 protein, which can form the basis for future investigation on the design of rapid diagnostic tests and more effective vaccine candidates for leptospirosis. [ABSTRACT FROM AUTHOR]

In this study, the potential of N-trimethyl chitosan (TMC) nanoparticles as a carrier system for the nasal delivery of the r4M2e.HSP70c, as an M2e-based universal recombinant influenza virus vaccine candidate, was investigated in mice. The anti-M2e specific cellular and humoral immune responses were assessed and the protective efficacy against a 90% lethal dose (LD90) of influenza A/PR/8/34 (H1N1) in a mice model was evaluated. Our results showed that the intranasal immunization of mice with r4M2e.HSP70c+TMC rather than the control groups, r4M2e+TMC, r4M2e and PBS (Phosphate buffer saline), significantly elevated both longevity and serum level of the total M2e-specific IgG antibody with a significant shift in the IgG2a/IgG1 ratio toward IgG2a, induced a Th1 skewed humoral and cellular immune responses, increased IFN-γ, IgG, and IgA in the bronchoalveolar lavage fluid (BALF), and promoted the proliferation of peripheral blood lymphocytes with lower morbidity and mortality rate against viral challenge. In conclusion, based on evidence to our finding, nasal vaccination with r4M2e.HSP70c antigen encapsulated into N-Trimethyl Chitosan (TMC) nanoparticulate system showed to induce a long lasting M2e-specific humoral and cellular immune responses and also provided full protection against a 90% lethal dose (LD90) of the influenza virus A/PR/8/34 (H1N1). It seems, protective immunity following intranasal administration of r4M2e could be resulted by the cooperation of both adjuvants, TMC and HSP70c. [ABSTRACT FROM AUTHOR]

As the importance of virus-specific IgG2a and strong induction of Th1 type immune response for virus clearance was reported, conventional influenza vaccines induce a highly humoral immune response and fail to induce cytotoxic T-lymphocyte (CTL) immunity. Hence, in agreement with heat shock protein 70 (HSP70) acting as Th1 cytokine-like adjuvant, an Escherichia coli -expressed r4M2e.HSP70c fusion protein comprising C-terminus of Mycobacterium tuberculosis HSP70 genetically fused to four tandem repeats of influenza A virus M2e was constructed. Then, the case-control study was carried out to evaluate the humoral and cellular responses elicited against M2e in Balb/C mice by intramuscular immunization with r4M2e.HSP70c alone. Our results showed that r4M2e.HSP70c rather than control groups, r4M2e, r4M2e + Alum, or rHSP70c, significantly elevated both longevity and serum level of the total M2e-specific IgG antibody, induced a Th1 skewed humoral and cellular immune responses, increased the level of IFN-γ in BALF, and promoted the proliferation of peripheral blood lymphocytes. Furthermore, a virus challenge experiment revealed that mice vaccinated with r4M2e.HSP70c limited the severity of influenza A disease by 100% survival rate, less sever body weight loss and delaying the onset of morbidity in mice for 2 days rather than other control groups. Here, we used r4M2e.HSP70c to stimulate M2e-specific antibody and cellular immune responses in Balb/C mice. The mHSP70c in the fusion form induced a long lasting Th1 skewed humoral and cellular immune responses against its associated protein. It seems anti-M2e antibodies limit viral replication and ameliorate influenza infection that allows the immune system to induce sterilizing HA-antibody against whole virion that leads to full protection against virulent influenza infection. [ABSTRACT FROM AUTHOR]

The effectiveness of inactivated H9N2 influenza vaccines is doubtful due to changes in antigenic regions of the virus hemagglutinin (HA) protein. One strategy for the development of the efficacious vaccine is the use of nanoparticles that display more immunogenic regions of the influenza virus. In this study, chitosan (CS)-based nanoparticles were developed as a delivery system for intranasal immunization using recombinant H9N2 virus HA1 and nucleoprotein (NP), for the induction of humoral and cellular responses. CS-HA1 and CS-NP nanoparticles were prepared by the ionic gelation method and characterized for their physicochemical properties and shape. The immunogenicity and the protective efficacy were evaluated by measuring antibody titers, T cell proliferation response, CD4+/CD8+ ratio, and quantitative real-time RT-PCR following intranasal administration of the prepared nanoparticles alone or in combination in chickens compared to an inactivated H9N2 vaccine. The average size, surface charge, and spherical structure of the synthesized nanoparticles showed high quality. Serologic analysis revealed that the immunization of inactivated vaccine groups resulted in strong influenza antibodies, which were significantly (p < 0.05) higher compared to the other groups. The vaccinated chickens with CS-HA1+CS-NP developed higher specific anti-influenza antibodies than in those vaccinated with each of rHA1 and rNP. Administration of a combination of the protein-based nanoparticles has stimulated the activation of both CD4+ and CD8+T cells and induced a significantly higher T cell proliferation. The viral shedding was significantly lower in CS-HA1+CS-NP and inactivated vaccine groups compared with other challenged groups. The data demonstrate the potential of CS-HA1+CS-NP nanoparticles for eliciting specific influenza antibodies and conferring protection in chickens. [ABSTRACT FROM AUTHOR]

As cellular immunity is essential for virus clearance, it is commonly accepted that no adequate cellular immunity is achieved by all available inactivated HA-based influenza vaccines. Thus, an improved influenza vaccine to induce both humoral and cell-mediated immune responses is urgently required to control LPAI H9N2 outbreaks in poultry farms. M2e-based vaccines have been suggested and developed as a new generation of universal vaccine candidate against influenza A infection. Our previous study have shown that a prime-boost administration of recombinant 4 × M2e.HSP70c (r4M2e/H70c) fusion protein compared to conventional HA-based influenza vaccines provided full protection against lethal dose of influenza A viruses in mice. In the present study, the immunogenicity and protective efficacy of (r4M2e/H70c) was examined in chickens. The data reported herein show that protection against H9N2 viral challenge was significantly increased in chickens by injection of r4M2e/H70c compared with injection of conventional HA-based influenza vaccine adjuvanted with MF59 or recombinant 4 × M2e (r4M2e) without HSP70c. Oropharyngeal and cloacal shedding of the virus was detected in all of the r4M2e/H70c vaccinated birds at 2 days after challenge, but the titer was low and decreased rapidly to reach undetectable levels at 7 days after challenge. Moreover, comparison of protective efficacy against LPAI H9N2 in birds intramuscularly immunized with r4M2e/H70c likely represented the ability of the M2e-based vaccine in providing cross-protection against heterosubtypic H9N2 challenge and also allowed the host immune system to induce HA-homosubtype neutralizing antibody against H9N2 challenge. This protective immunity might be attributed to enhanced cell-mediated immunity, which is interpreted as increased lymphocytes proliferation, increased levels of Th1-type (IFN-γ) and Th2-type (IL-4) cytokines production and increased CD4 + to CD8 + ratios, resulting from the injection of four tandem repeats of the ectodomain of the conserved influenza matrix protein M2 (4 × M2e) genetically fused to C-terminus of Mycobacterium tuberculosis HSP70 (mHSP70c). [ABSTRACT FROM AUTHOR]

Utilizing subunit vaccines is one of the strategies to address influenza infection. Recent innovations have focused on high doses of vaccine antigens and immune enhancers or adjuvant to induce more robust and long-lasting immune responses. Here, an effect of the B cell-activating factor receptor (BAFF-R) to increase the magnitude and durability of immune responses of the recombinant HA1 (rHA1) protein against the H1N1 influenza virus was studied. The HA1 protein and the effector domain of BAFF-R were expressed in the pET-28a (+) vector. Eight-week-old BALB/c mice were equally grouped into five groups (n=20). The 15 and 25 μg/μL of rHA1 were mixed with 2 μg/μL of rBAFF-R and injected three times for vaccinated groups. Three control groups were received normal saline and two concentrations of rHA1. The ability of rBAFF-R in eliciting HA-specific antibody response and stimulating T lymphocyte proliferation to induce the cell-mediated immunity was assayed. Induction of protection was evaluated following the challenge with PR8 strain. Analysis of immune responses showed that the co-administration of rBAFF-R with rHA1 boosted HI responses to the antigen in mice, whilst it was not able to promote the T cell proliferation responses against influenza. Compared to rHA1alone, the rBAFF-R/rHA1 generated efficient protection for the animals. There were no significant differences in eliciting the immune responses in mice immunized with the lower dose of rHA1 than that with the higher dose. The data indicate the rBAFF-R can enhance the primary and memory immune responses to protect against influenza infection. [ABSTRACT FROM AUTHOR]

Abstract: Ideal vaccines against influenza viruses should elicit not only a humoral response, but also a cellular response. Mycobacterium tuberculosis HSP70 (mHSP70) have been found to promote immunogenic APCs function, elicit a strong cytotoxic T lymphocyte (CTL) response, and prevent the induction of tolerance. Moreover, it showed linkage of antigens to the C-terminus of mHSP70 (mHSP70c) can represent them as vaccines resulted in more potent, protective antigen specific responses in the absence of adjuvants or complex formulations. Hence, recombinant fusion protein comprising C-terminus of mHSP70 genetically fused to four tandem repeats of the ectodomain of the conserved influenza matrix protein M2 (M2e) was expressed in Escherichia coli, purified under denaturing condition, refolding, and then confirmed by SDS–PAGE, respectively. The recombinant fusion protein, 4xM2e.HSP70c, retained its immunogenicity and displayed the protective epitope of M2e by ELISA and FITC assays. A prime-boost administration of 4xM2e.HSP70c formulated in F105 buffer by intramuscular route in mice (Balb/C) provided full protection against lethal dose of mouse-adapted H1N1, H3N2, or H9N2 influenza A isolates from Iran compared to 0–33.34% survival rate of challenged unimmunized and immunized mice with the currently in use conventional vaccines designated as control groups. However, protection induced by immunization with 4xM2e.HSP70c failed to prevent weight loss in challenged mice; they experienced significantly lower weight loss, clinical symptoms and higher lung viral clearance in comparison with protective effects of conventional influenza vaccines in challenged mice. These data demonstrate that C-terminal domain of mHSP70 can be a superior candidate to deliver the adjuvant function in M2e-based influenza A vaccine in order to provide significant protection against multiple influenza A virus strains. [Copyright &y& Elsevier]

Abstract: There is a growing concern regarding continuous risk of emerging a new influenza pandemic. It is highlighted the need for novel vaccination techniques that quickly and effectively employed to respond to such threats. Although, DNA vaccine is a simple and effective approach to induce antigen specific immune responses, their potency requires further improvement. DNA vaccine encoding conserved antigen of influenza virus could provide protection in various animal models. Therefore, we constructed a plasmid vector encoding M2e-HSP70c sequences, pcDNA/MHc, as a candidate for universal influenza vaccine. The expression of newly constructed vectors was verified by transient transfection of mammalian cells (HEK293T cell line) and western blot analysis using commercial antibodies. Mice were injected subcutaneously (s.c.) by the help of electroporation (IEP) in the footpad area and boosted without IEP with 100μg of constructed vector. Furthermore, the potency of this construct to provoke humoral immune responses and its protectivity against lethal dose of viral challenge were evaluated. Based on our study, the fusion construct was immunogenic in mice and was able to confer both protection against lethal challenge of H1N1 virus and reduce viral load in lung homogenates of the infected mice. [Copyright &y& Elsevier]

Objective(s): Multiple Sclerosis is a central nervous system disease which belongs to the category of autoimmune diseases. The prevalence of this disease in Iran is approaching the European level. Astrocyte cells are nerve tissues that regulate the immune system activity by secreting various cytokines such as IL- 17. The aim of this study was partial purification of toxin from M.eupeus scorpion venom that has immunomodulatory effect on astrocyte cell line (1321N1) Materials and Methods: In the present study, purified crude venom of M.eupeus scorpion. Size exclusion and reverse-phase highperformance liquid chromatography was used for fractionation. The fractional molecular weight was determined by Using SDS and Tricine electrophoresis, Astrocyte cells (1321N1) were selected as functional cells in testing the immunomodulatory effect of venom. The viability of cells were determined by MTT and LDH assays. Astrocyte cells were activated by lipopolysaccharide and the release of interleukin-17 in activated cells was estimated using ELISA kit. Results: fraction 331 (F331) from RP-HPLC contain the purified peptide with molecular weight of about 4500 Dalton. When activated cells exposed to purified peptide the rate of interleukin-17 release was found to be 85 pg/ml which is almost similar to un-activated cells (78 pg/ml). However in activated cells by LPS without treatment with purified peptide the rate of IL-17 release was found to be 147 pg/ml which was significantly (P<0.05) higher than control group. Conclusion: The purified peptide (F331) from venom of Mesobouthus eupeus can inactivate the astrocyte 1321N1 cells activated by LPS as indicated by decreased secretion of IL-17 from the cells. [ABSTRACT FROM AUTHOR]

The mucosal immune system serves as the first line of defense against Bordetella pertussis. Intranasal vaccination, due to its potential to induce systemic and mucosal immune responses, appears to prevent the initial adherence and colonization of the bacteria at the first point of contact. In the present study, two B. pertussis antigens, pertussis Toxoid (PTd) and Filamentous hemagglutinin (FHA), which play a very significant role in virulence and protection against pertussis, were encapsulate into N-trimethyl chitosan (TMC) nanoparticulate systems. After preparation of TMC nanoparticles (NPs), the NPs were characterized and their ability to induce efficient immune responses against B. pertussis was studied in a mouse model. Our findings showed that PTd + FHA-loaded TMC NPs have strong ability to induce IL-4, IL-17, IFN-γ, IgG, and IgA in the mouse model. Results from this study suggest that nasal administration of the PTd + FHA-loaded TMC NPs induced not only a systemic immune response but also a local mucosal response, which may improve the efficacy of pertussis prevention through respiratory tract transmission. [ABSTRACT FROM AUTHOR]

Background and Objectives: Antimicrobial peptides produced by lactic acid bacteria have gained enormous attention owing to their health benefits. This study aimed to isolate, purify and characterize the antibacterial protein produced by autochthonous Lactobacillus casei TA0021 strain. Materials and Methods: The antagonistic activity of L. casei TA0021 against a number of pathogenic bacteria was tested by agar well diffusion assay. The antimicrobial agent in the neutralized supernatant fluids was subjected to the action of proteolytic enzymes, catalase, lipase and lysozyme, and their tolerance to variable pH and temperature was estimated. The proteinaceous antagonistic compound was precipitated by 60% w/v ammonium sulphate, desalted and subjected to cation exchange and gel filtration chromatography. Approximate molecular weight of Lactocin was determined by SDS-PAGE and non-denaturing gel electrophoresis. Hemoglobin release assay and cytotoxicity effect of Lactocin TA0021 was determined. The results were statistically analyzed. Results: The antagonistic agent active against Salmonella Typhimurium and Shigella flexneri appeared resistant to catalase and lipase treatments, while sensitive to the tested proteolytic enzymes. Lactocin TA0021 resisted acidic pH values of 3.0, while alkaline pH values of >9 completely destroyed the activity. The antibacterial peptide was approximately 68 KDa and heat labile as lost its activity at 100°C after 5 minutes. The bacteriocin was non-toxic to MRC-5 cell lines and non-hemolytic. Purification method lead to increase in antibacterial activity while, subsequent decrease in recovery and yield was observed with increasing purification fold. Conclusion: The purified antimicrobial protein from L. casei TA0021 might be used for application in medicinal and food products. [ABSTRACT FROM AUTHOR]

Background CD4+ CD25+ T-regulatory ( Treg) cells play critical roles in maintaining peripheral tolerance and preventing autoimmunity. As characteristics of Treg cells have not been precisely investigated in chronic urticaria ( CU) yet, this study was performed. Objective To determine the frequencies of circulating CD4+ CD25+ FOXP3+ T cells and serum levels of interleukin ( IL)-10, transforming growth factor ( TGF)- β, and IL-17 in patients with chronic autoimmune urticaria and chronic idiopathic urticaria compared to healthy controls. Methods Peripheral blood mononuclear cells ( PBMCs) were obtained from patients with CU and healthy controls. The frequency of CD4+ CD25+ T cells in PBMCs and expression levels of FOXP3 were detected by flow cytometry. The serum levels of IL-10, TGF- β, and IL-17 were measured by enzyme-linked immunosorbent assay. Results A significant decrease in the percentage of circulating CD4+ CD25+ FOXP3+ T cells was detected in patients with CU, compared to control subjects. However, no significant difference was detected on the serum levels of IL-10, TGF- β, and IL-17 between patients with CU and control subjects. Conclusions This study demonstrated that the frequency of Treg cells in PBMCs was decreased in patients with CU. Further studies are needed to clarify the exact role of Treg cells in the pathogenesis of CU and factors regulating their function. [ABSTRACT FROM AUTHOR]

Invasive aspergillosis increases in chronic immunosuppressive diseases such as cancer. There is little information about the mechanisms by which Aspergillus infection affects the immune regulation and microenvironment of cancer cells. Hence, this study was aimed at investigating the effect of invasive aspergillosis on immunosurveillance, metastasis, and prognosis of cancer in tumor-bearing mice. After implantation of mouse mammary tumor in BALB/c mice, they were infected with Aspergillus conidia intravenously. For comparison, groups of mice were experimentally infected with Aspergillus conidia or implanted with tumor cells separately. Seven days after Aspergillus infection, the serum levels of tissue inhibitor of metalloproteinase-1 (TIMP-1) were measured by ELISA, and subsequently regulatory T lymphocytes were analyzed by flow cytometry. The survival of animals and mean tumor size were then determined. Our results indicated that tumor sizes in mice increased significantly after infection with Aspergillus conidia. Moreover, invasive aspergillosis enhanced the population of regulatory lymphocytes and level of TIMP-1. This study supports the idea that massive Aspergillus infection could stimulate tumor growth and increases the possibility of a bad prognosis. As a result, treatment of Aspergillus infection could be considered an important issue for efficient cancer therapy. Les maladies immunosuppressives chroniques comme le cancer augmentent l'incidence d'aspergillose invasive. Il n'y a que peu d'informations disponibles sur les mécanismes par lesquels l'infection à Aspergillus affecte la régulation immune et le microenvironnement des cellules cancéreuses. Ainsi, cette étude visait à examiner l'effet de l'aspergillose invasive sur l'immunosurveillance, la métastase et le pronostic du cancer chez des souris porteuses de tumeurs. Après l'implantation de tumeurs mammaires chez la souris BALB/c, celles-ci ont été infectées par des conidies d'Aspergillus de façon intraveineuse. Pour fins de comparaison, des groupes de souris ont été séparément infectés par des conidies d'Aspergillus ou implantés avec des cellules tumorales. Sept jours après l'infection à Aspergillus, les niveaux sériques de TIMP-1 (tissue inhibitor of metalloproteinase-1) ont été mesurés par ELISA et les lymphocytes T régulateurs (Treg) ont été analysés subséquemment par cytométrie de flux. La survie des animaux et la taille moyenne des tumeurs ont ensuite été déterminées. Nos résultats indiquent que la taille des tumeurs des souris augmentait significativement à la suite de l'infection par les conidies d'Aspergillus. Parallèlement, l'aspergillose invasive augmentait la population de lymphocytes régulateurs et le niveau de TIMP-1. Cette étude appuie l'idée que l'infection massive par Aspergillus puisse stimuler la croissance tumorale et détériorer le pronostic. Conséquemment, le traitement de l'infection à Aspergillus devrait être considéré comme un problème important à prendre en compte pour l'efficacité de la thérapie anticancéreuse chez les patients. [ABSTRACT FROM AUTHOR]