ABSTRACT
Tuberculosis is a disease with high morbidity, caused mainly by Mycobaterium tuberculosis [M.tb.]. DNA vaccines show a promising future due to their unique advantages over conventional methods. The early-secreted antigen target [ESAT]-6 and culture filtrate protein [CFP]-10 of M.tb. antigens have been identified as vaccine candidates against Mycobacteria and used as subunit vaccines, DNA or protein, in different studies. To investigate the potential of pcDNA3.1+ plasmid containing CFP-10 and ESAT-6 genes in induction of local immune responses after intramuscular injection in BALB/c mice. pcDNA 3.1+ CFP-10 and pcDNA3.1+ ESAT-6 plasmids were prepared and defined groups of mice were injected intramuscularly with the plasmids both separately and in combination. The RNA was extracted from muscles after one month and cDNA was made using RT-PCR. The expressions of IL-4, IL-10 and IFN-gamma genes cytokines were evaluated using comparative real time PCR. Expression of IL-4 and IL-10 increased in the injection site of the mice groups which received plasmids encoding ESAT-6 and CFP-10 individually or together. More than 10-fold increase in IFN-gamma expression was found in samples taken from mice groups inoculated by plasmids encoding ESAT-6 and CFP-10 individually or together. pcDNA 3.1+ESAT-6 and pcDNA3.1+CFP-10 plasmids can increase the expression of IFN-gamma in mice after immunization.
ABSTRACT
DNA vaccines are third generation vaccines which have made promises to combat infectious diseases. Cationic liposomes are used as effective delivery systems for DNA vaccines to generate stronger immunity. Encapsulation of pcDNA3.1+PA plasmid, encoding protective antigen [PA] of Bacillus anthracis [B. anthracis] into cationic liposomes, and evaluation of its effect on specific humoral specific immunity against PA were aimed. The liposomes containing pcDNA3.1+PA plasmids were prepared with phosphatidylcholine [PC], dioleoyl phosphatidylethanolamine [DOPE] and 1,2-dioleoyl-3-trimethylammonium-propane [DOTAP] using dehydration-rehydration method. BALB/c mice were immunized by intramuscular [IM] injection to investigate the immunogenicity of the formulations. The resulting specific antibodies against PA, total IgG, IgG1, IgG2a and IgG2b isotypes, were evaluated by enzyme linked immunosorbent assay [ELISA] method. A higher concentration of specific IgG against PA was found in sera of a group immunized with the encapsulated plasmid compared with the naked plasmid alone. This difference was significant for IgG1 isotype
ABSTRACT
Saffron [Zaaferan], botanical name Crocus sativus, is the most expensive spice in the world. It is derived from the dried stigma and pistil of the purple saffron crocus flowers. Iran is the largest saffron producer accounting for more than 80% of the world's production. Saffron contains an aeroallergen that causes reactive respiratory allergic reactions in atopic subjects. IgG antibody to allergens in the serum of allergic patients is not routinely measured. In this study in order to find out more about mechanism of allergy against saffron pollen, specific antibodies [IgE and IgG, total and subclasses] in atopic subjects were assayed. We used an ELISA assay for measuring specific IgE and IgG against saffron pollen extract in the sera of 38 atopic subjects [test group] and 20 non allergic subjects [control group]. The optical densities were compared between allergic subjects and non-allergic individuals. The prick test with saffron pollen extract was used to evaluate the cutaneous and specific antibody responses in the allergic subjects. The correlation was determined by statistical analysis. Specific saffron pollen IgE and IgG subclasses were found significantly higher in the allergic subjects than the control group. The immediate skin reaction was found positive in 70% of the test group. We report here, the existence of a positive correlation between specific IgE and skin reaction by prick test in atopic subjects [R=0.433]. A negative correlation between specific IgE and IgG4 subclass was also found [R=-0.576]. These data may be useful to understand the mechanism of allergy to saffron and may help in clarifying clinical manifestations and to prevent IgE production as well as therapeutic application
ABSTRACT
DNA immunization with plasmid DNA encoding bacterial, viral, parasitic and tumor antigens has been reported to trigger protective immunity. To evaluate the use of a DNA immunization strategy for protection against anthrax, a plasmid was constructed. The partial sequence of protective antigen of Bacillus anthracis, amino acids 175-764, as a potent immunogenic target was selected. The DNA encoding this segment was utilized in the construction of pcDNA3.1+PA plasmid. After intramuscular injection of rats with pcDNA3.1+PA plasmid, the expression of PA was assessed by RT-PCR and immunohistochemistry at RNA and protein levels, respectively. We also evaluated the presence of anti-PA antibodies in sera of immunized mice with pcDNA3.1+PA construct using immunoblotting. The integrity of pcDNA3.1+PA construct was confirmed with restriction analysis and sequencing. The expression of PA was detected at RNA and protein levels. The presence of anti-PA antibodies in immunized mice with pcDNA3.1+PA construct was also confirmed. Our results indicate that pcDNA3.1+PA eukaryotic expressing vector could express PA antigen, induce antibody response and may be used as a candidate for DNA vaccine against anthrax
Subject(s)
Animals, Laboratory , Eukaryotic Cells , Bacillus anthracis/genetics , Plasmids , ImmunohistochemistryABSTRACT
Background: Vitiligo is a dermatological disorder of unknown etiology with a common incidence in southern Iran. Presence of autoantibodies to melanocyte antigens suggested an autoimmune basis of the disease
Objective: In this study, the presence of rheumatoid factor [RF] in sera and skin biopsies of vitiligo patients was investigated
Methods: The presence of RF in sera of 35 vitiligo and 32 normal individuals was assessed by an indirect ELISA assay. In addition, the presence of IgM, IgG, and IgA immunoglobulins in the biopsy lesions of patients was also investigated by Immunoperoxidase test
Results: IgM-RF and IgA-RF were detected in sera of 50% and 20% of patients, respectively. Five out of 35 [15%] revealed to produce both IgM and IgA rheumatoid factors. The rheumatoid factor activity of the deposited immunoglobulins at the site of lesion was confirmed by direct immunoperoxidase test
Conclusion: The presence of rheumatoid factors as non organ-specific autoantibodies in vitiligo provides further evidence for the autoimmune etiology of the disease and its pathological importance remains to be elucidated