Development of a Gene Therapy Method for Cervical Cancer Using Attenuated Coxsackievirus B3 as a Vector System

We previously reported the development of an attenuated coxsackievirus B3, known as YYFF, which functioned as a viral vector system for foreign gene expression. In this study, we demonstrated the potential use of YYFF as a gene therapy vector. Recombinant YYFF was constructed to express the human papillomavirus 16 (HPV16) E7 gene, referred to as YYFF-HPV16-E7. Growth of YYFF-HPV16-E7 resembled the wild type, YYFF, and it expressed HPV16-E7 in cell culture. When YYFF-HPV16-E7 was directly injected into TC-1-transplanted C57/BL6 mice, there was no reduction in tumor size, because of the non-growth of YYFF in C57/BL6 mice. However, when YYFF-HPV16-E7-induced immune cells/serum that originated from BALB/c mice was passively delivered into BALB/c background TC-1-transplanted nude mice, it reduced the size of cervical tumors in the nude mice. This study indicates the potential use of YYFF-HPV16-E7 as a gene therapy agent for treating HPV-induced cervical cancer.

Neutralizing Antibody Induction and Cytotoxic T Lymphocyte Response to Nakayama-NIH and Beijing-1 as Japanese Encephalitis Virus Vaccine Strains

The Japanese encephalitis virus (JEV), a member of the Flaviviridae family and Flavivirus genus, is transmitted by mosquitoes. JEV, of which some 35,000 cases are recorded every year, is a positive RNA virus. Two types of JEV vaccines have been developed to prevent the onset of encephalitis in humans, namely formalin-inactivated and liveattenuated vaccines. JEV inactivated vaccines are usually made using the Nakayama-NIH or Beijing-1 strains of the JEV virus. In this study, the immunological response to the Nakayama-NIH and Beijing-1 strains was analyzed as part of the effort to compile basic data which could lead to the selection of a suitable vaccine strain. To this end, the virus titer of Beijing-1 was found to be two-fold higher than that of Nakayama-NIH by plaque assay. Moreover, Beijing-1-induced neutralizing antibodies showed a higher level of titers when confronted by Korean JEV isolates than Nakayama-NIH-induced neutralizing antibodies (1:320 vs. 1:160, respectively). However, as a minimum ratio of 1:10 neutralizing antibody titers are required to protect against JEV infection, both strains in effect exhibited a sufficient level of neutralizing antibody titers. What's more, Beijing-1 was found to induce a somewhat higher cytotoxic T lymphocyte (CTL) response than Nakayama-NIH. Taken together, this can be taken to mean that Beijing-1 may in fact be a more effective vaccine candidate strain when it comes to inducing a high level of protective immunity against JEV infection.

Host Gene Profiling of Coxsackievirus B3 H3- and 10A1-infected Mouse Heart

Coxsackievirus B3 (CVB3) is a non-enveloped virus that has a single-stranded RNA genome. CVB3 induces myocarditis, and ultimately, dilated cardiomyopathy. A myocarditis variant of CVB3 (CVB3 H3) and its antibody-escape mutant (CVB3 10A1) were studied previously; H3 was found to induce myocarditis and 10A1 was found to be attenuated in infected mice. Although amino acid residue 165, located in a puff region of VP2, was found to be altered (i.e., the H3 asparagine was altered to aspartate in 10A1), the detailed mechanism of attenuation was not clearly elucidated. Here, DNA microarray technology was used to monitor changes in mRNA levels of infected mouse hearts after CVB3 H3 and 10A1 infection. This tool was used to elucidate the pathogenic mechanisms of viral infection by understanding virus-host interactions. We identified several genes, including protein tyrosine kinases, Ddr2 and Ptk2, as well as Clqb and Crry, involved in complement reactions, which may be involved in these viral processes. Thus, gene profiling can provide an opportunity to understand host immune responses to viral infection for gene therapy and may contribute to the identification of the target gene that is modified during treatment of viral myocarditis.

Development of Peptide Antibody against Coxsackievirus B3 VP2

Coxsackievirus B3 (CVB3) is the nonenveloped virus containing a single-stranded positive-sense RNA as a genome. CVB3 infection can induce acute myocarditis and dilated cardiomypathy. CVB3 of icosahedral symmetry has four capsid proteins called VP1, VP2, VP3, and VP4. Although VP1 is a major antigenic determinant, VP2 is also an important protein for viral physiology, such as maturation cleavage and attenuation. However, VP2 study has been hampered, partly because VP2 antibody is not available. In this study, we developed peptide-based polyclonal VP2 antibody and analyzed its potency by Western blotting analysis and immunofluorescent assay. Purified B3-1 antibody (VP2 peptide antibody developed in here) showed the sensitivity and specificity, similar to VP1 monoclonal antibody which is commercially available. Moreover, this peptide antibody may be useful for double-staining with other antibodies derived from mouse. Therefore, the VP2 antibody may allow us to study CVB assembly and understand VP2 function in depth.

A Case of Huge Cystic Lymphangioma of the Greater Omentum

Lymphangioma is a tumor which is composed of lymph vessels and spaces containing lymph. It consists of endothelial cells and supporting tissue elements of the lymphatic system which are involved in the neoplastic process. Grossly, the cysts are thin walled and multiloculated. Intra-abdominal omental origin lymphangiomas are quite rare and only 21 cases were reported in English literature by 1978. The most common location is the mesentery, followed by the omentum, mesocolon, and retroperitoneum. We report a case of huge cystic lymphangioma originating from the greater omentum in a 14-year-old male patient with brief review of literatures.

Hematogenous Candida Pneumonia in Major Burn Patients: Plain Chest Radiograph and Thin-section CT Findings

PURPOSE: To describe plain radiographic and thin-section CT findings of hematogenous candida pneumonia in major burn patients. MATERIAL AND METHOD: We reviewed nine cases of hematogenous candida pneumonia in major burn patients who had positive blood culture for candida and findings of pneumonia on plain chest radiograph. On five of nine cases, thin-section CT was done. We evaluated retrospectively nine cases for onset, the pattern, distribution, and size of lesions on plain chest radiograph and thin-section CT. RESULTS: On plain chest radiograph, randomly distributed 2-10mm nodules were seen in six cases(66%) and randomly distributed 10-15mm consolidations in remaining three cases{33% ). Lesion occured in 11th to 75th post-burn day{average, 34th post-burn day). Other findings were cardiomegaly in three cases, atelectasis in three cases, and pulmonary edema in one case. Thin-section CT showed variable shaped subpleural nodules in all five cases. The size of nodules were 1-5mm in two cases(40%) and 5-10mm(60% ) in three cases. Feeding vessel signs were seen in two cases. Other findings were atelectasis in three cases, cardiomegaly in three cases, ground-glass opacity and interlobular septal thickenings by pulmonary edema in two cases. CONCLUSION: Plain chest radiographic findings of hematogenous candida pneumonia in major burn patients are randomly distributed nodules or consolidations of variable size. Thin-section CT findings are variable shaped subpleural nodules less than 1 cm.