فهرست مطالب فاضل شکری

Hepatitis B, as a major healthcare problem and a potentially life-threatening infection, is caused by Hepatitis B virus (HBV). Nucleocapsid of HBV is composed of a single polypeptide chain known as core antigen, HBcAg. As a highly immunogenic subviral particle, HBcAg has been used as a carrier platform for heterologous proteins. Anti-HBc antibody, as one of the serological markers for HBV infection, is the best marker for documenting prior exposure to HBV. Here, we produced, purified and characterized HBcAg. pColdI vector, containing a histidine tag, was used for the first time to clone HBcAg gene. The highest level of HBcAg expression was achieved in Tuner strain, followed by BL21 and Rosetta-gami. Histidine-tagged HBcAg was purified using Ni-NTA resin, and its structural and antigenic similarity with natural HBcAg was confirmed by ELISA and western blot. Then, recombinant HBcAg was used to produce anti-HBc polyclonal antibodies in rabbits, and specific anti-HBcAg polyclonal antibodies were purified by the HBcAg-CNBr sepharose column. Specific anti-HBcAg polyclonal antibodies were able to detect recombinant HBcAg up to 0.15 ng/ml in an optimized sandwich ELISA. The results of this study provide valuable information for the production of recombinant HBcAg and specific anti-HBcAg polyclonal antibodies for further research and diagnostic applications.

To detcnninc the short-term reacrogenicity of diphtheria-tetanus-whole cell pertussis (DTwP) vaccine administered to preschool children in a number of health centers of Tehran in 2006.

In this prospective cohort study, 337 children aged 4-6 years were injected with DTwP vaccine manufactured by Razi Institute of Iran. Reactogenicity was assessed by the parents for 7 days post vaccination using diary cards. Local (pain, redness and swelling) and systemic (fever, loss of appetite, gasterointestinal symptoms, vomiting and eczema) side effects were recorded daily.

Out of 337 children, 312 (92.6%) reported local reactions and 220 (65.3%) reported systemic reactions. No serious adverse events related to vaccination were reported. Among local reacuons, pain was the most frequent (86.9%), but 11 was mostly mild or moderate. Redness (52.8%) and swelling (41.2%) were the most frequently observed signs in the second day. The systemic reactions observed in children included fever (48.4%), loss of appetite (24%), gastrointestinal symptoms (5 6%), vomiting (8%) and eczema (2.7%). Only 3.6% of children had auxiliary fever above 39 'c. All signs were observed to have reduced or completely disappeared during a week.

Compared with previous reports in Iran, reactogenicity of DTwP of Razi Institute seems to be reduced, but it was still more frequent than the internationally approved cellular vaccine counterparts. Reactogcnicity of the cellular triple vaccine may be related to the vaccine formulation or the bacterial cell fragments used in vaccine production.

As monoclonal antibodies are potential tools for characterization of soluble or cellular surface antigens, use of these proteins has always been considered in infertility and reproduction research. Therefore, in this study, monoclonal antibodies against human sperm surface antigens were produced.

To produce specific clones against human sperm surface antigens, proteins were extracted using solubilization methods. Balb/c mice were immunized intraperitoneally with the proteins using complete Freund’s adjuvant in the first injection and incomplete Adjuvant in the following booster injections. Hybridoma cells producing ASA were cloned by limiting dilution.

Five stable ASA producing hybridoma clones were achieved and their antibody isotypes were determined by ELISA. All the isotypes were of IgG class. Their cross reactivity with rat and mice spermatozoa was examined but they did not have any cross reactivity.

The produced antibodies can be used in further studies to characterize and evaluate each of the antigens present on human sperm surface and determining their role in fertilization.