Immunization with Adenoviral Vectors Carrying Recombinant IL-12 and E7 Enhanced the Antitumor Immunity against Human Papillomavirus 16-associated Tumor / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Human papillomavirus (HPV) infection has a significant role in cervical carcinogenesis, and HPV oncoprotein E7 plays an important part in the formation and maintenance of cervical cancer. Interleukin-12 (IL-12) has been reported to induce a cellular immune response, and to suppress the tumor growth and the E7 production. Here we describe the use of adenoviral delivery of the HPV 16 E7 subunit (AdE7) along with adenoviral delivery of IL-12 (AdIL-12) in mice with HPV-associated tumors. MATERIALS AND METHODS: Mice were injected with TC-1 cells to establish TC-1 tumor, and then they were immunized with AdIL-12 and/or AdE7 intratumorally. The anti tumor effects induced by AdIL-12 and/or E7 were evaluated by measuring the size of the tumor. E7-specific antibody and INF-gamma production in sera, and the T-helper cell proliferative responses were then measured. Cytotoxic T-lymphocyte (CTL) and T cell subset depletion studies were also performed. RESULTS: Combined AdIL-12 and AdE7 infection at the tumor sites significantly enhanced the antitumor effects more than that of AdIL-12 or AdE7 single infection. This combined infection resulted in regression of the 9 mm sized tumors in 80% of animals as compare to the PBS group. E7-specific antibody and INF-gamma production in the sera, and the T-helper cell proliferative responses were significantly higher with coinfection of AdIL-12 and AdE7 than with AdIL-12 or AdE7 alone. CTL response induced by AdIL-12 and AdE7 in the coinjected group suggested that tumor suppression was mediated by mostly CD8+ and only a little by the CD4+ T cells. CONCLUSION: IL-12 and E7 application using adenovirus vector showed antitumor immunity effects against TC-1 tumor, and this system could be use in clinical applications for HPV-associated cancer.

Photodynamic Effects of Radachlorin(R) on Cervical Cancer Cells / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Photodynamic therapy (PDT) is a novel treatment modality, which produces local tissue necrosis with laser light following the prior administration of a photosensitizing agent. Radachlorin(R) has recently been shown to be a promising PDT sensitizer. In order to elucidate the antitumor effects of PDT using Radachlorin(R) on cervical cancer, growth inhibition studies on a HPV-associated tumor cell line, TC-1 cells in vitro and animals with an established TC-1 tumor in vivo were determined. MATERIALS AND METHODS: TC-1 tumor cells were exposed to various concentrations of Radachlorin(R) and PDT, with irradiation of 12.5 or 25 J/cm2 at an irradiance of 20 mW/cm2 using a Won-PDT D662 laser at 662 nm in vitro. C57BL/6 mice with TC-1 tumor were injected with Radachlorin(R) via different routes and treated with PDTin vivo. A growth suppression study was then used to evaluate the effects at various time points after PDT. RESULTS: The results showed that irradiation of TC-1 tumor cells in the presence of Radachlorin(R) induced significant cell growth inhibition. Animals with established TC-1 tumors exhibited significantly smaller tumor sizes over time when treated with Radachlorin(R) and irradiation. CONCLUSION: PDT after the application of Radachlorin(R) appears to be effective against TC-1 tumors both in vitro and in vivo.

Changes of allergenecity of salted and fermented shrimp / 천식및알레르기

BACKGROUND AND OBJECTIVE: Shrimp is one of the most common food allergens. The salted and fermented shrimp (S/F shrimp) is a traditional food and ingested frequently in the daily life of many Korean people. But few studies have been investigated on the allergenicity of S/F shrimp. The aim of our study is to observe the allergenicity of S/F shrimp and to compare it with the allergenicity of shrimp. MATERIAL AND METHODS: Crude extracts were made from three kinds of S/F shrimps and four kinds of shrimps after boiling. The extracts were used for SDS-PAGE, ELISA, ELISA inhibition test, immunoblotting, and immunoblotting inhibition test with sera from 10 subjects who have shrimp specific IgE by radioallergosorbent (RAST) (3+ or 4+). RESULTS: The protein concentrations of S/F shrimp extracts were less than half of the extracts of shrimps (1.9 mg/ml, 1.2 mg/ml, 1.1 mg/ml vs 4mg/ml, 3.83 mg/ml, 6.8 mg/ml, 6.6 mg/ml). The extracts of S/F shrimp showed weaker and more diffusely distributed protein bands in SDS-PAGE and showed less than half of the O.D. value in IgE-ELISA compared to those of shrimp extracts. The specific IgE immunoblotting of S/F shrimp extracts showed 9 bands (36, 33, 30, 29, 28, 26, 23, 20, 19 kDa). Dose dependent inhibition was observed between shrimps and S/F shrimp in ELISA-inhibition and immnunoblotting inhibition test. The allergen concentration of S/F shrimp needed to inhibit 50% of shrimp-lgE ELISA was five times higher than that of shrimp. CONCLUSION: The crude extracts of S/F shrimps showed less than half the amount of protein compared to crude extracts of shrimps have and also showed reduced allergenicity by salting and fermentation.

Expression and characterization of a mutant recombinant blood coagulation factor VIII (rFVIII (m))

In an earlier study, a site directed mutant rFVIII (rFVIII(m), Arg(336) -> Gln(336)) expressed in baculovirus-insect cell (Sf9) system was found to sustain high level activity during incubation at 37 for 24 h while the cofactor activity of normal plasma was declined steadily. In this study, a mutant B-domain deleted rFVIII(m), Arg(336) -> Gln(336) expressed in baculovirus-insect cell (Sf9) system was characterized for its enzymatic and chemical properties. The expressed rFVIII(m) and plasma FVIII (pFVIII) were purified by immunoaffinity column chromatography and identified by Western blot analysis. The partially purified rFVIII(m) exhibited cofactor specific activity of 2.01 X 10(3)units/mg protein. The molecular weight of rFVIII(m) ranged between 40 to 150 kDa with a major band at 150 kDa. Treatment of both rFVIII(m) and pFVIII with thrombin increased their cofactor activity in a similar pattern. Treatment of both the activated rFVIII(m) and native FVIII with APC decreased their cofactor activities, however, the former exhibited a slower decrease than the latter, although no significant difference was present. rFVIII(m) formed a complex with vWF, resulting in a stabilized form, and the lag period of thrombin-mediated activating was extended by vWF association. These results implicated that rFVIII(m) expressed in baculovirus-insect cell system had a comparable capacity as FVIII cofactor activity and might be a good candidate for the FVIII replacement therapy for hemophilia A patients.