• IMMUNE NETWORK
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• v.1 no.2
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• pp.109-115
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• 2001

Background: The role of the interferon consensus sequence binding protein (ICSBP), a member of interferon regulatory factor family, in protecting against a vesicular stomatitis virus (VSV) infection has not been firmly elucidated. Thus, it was investigated utilizing the human promyelocytic leukemia HL-60 cells which do not express ICSBP. Methods: HL-60 cells were stably transfected with plasmid containing cDNA for either ICSBP or DNA binding domain (DBD) and tested for their VSV-susceptibilities. The susceptibility of each transfectant group to a VSV infection was determined by a plaque assay at 1 h, 24 h, and 48 h post-infection in the presence (500 IU/ml) or absence of interferon ${\alpha}$ ($IFN{\alpha}$). Results: In the absence of $IFN{\alpha}$, the three groups showed similar sensitivities to a VSV infection. However, when pre-treated with IFN, the viral titers in both the ICSBP and control clones steadily decreased over 48 h of incubation, indicating the existence of $IFN{\alpha}$-mediated protection against VSV infection. The $IFN{\alpha}$-treated ICSBP clones appeared to be more resistant to infection compared with the control clones, although the difference was not great. On the contrary, the viral titers in the $IFN{\alpha}$-treated DBD clones increased at 24 h then decreased by 48 h. Conclusion: The expression of truncated ICSBP (DBD) does not appear to underlie the impaired protection against a VSV infection in the DBD clones, since even the control clones lacking ICSBP were protected from a VSV infection. This suggests that ICSBP does not play a critical role in the $IFN{\alpha}$- mediated anti-VSV response of HL-60 cells, although it appears to confer some resistance to a VSV infection.

• KSBB Journal
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• v.5 no.1
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• pp.49-58
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• 1990

We constructed hybrid plasmids to allow controlled and extracellular production of human alpha-interferon in Escherichia coli. The hybrid plassmids were constructed by transferring alpha-lFN gene from plasmid Hif-2h which has the alpha-lFN gene at PstI restriction site of pBR322, to plasmids pIN -IIIB3 and pIN-IIIC3 at restriction sites between HindIII and BamHI. Plasmids pIN-IIIB3 and pIN-IIIC3 carry E. coli lipoprotein promoter, lac promoter and operator in tandem. The plasmids also have lacl genes which encode for lac repressors, which allows controlled expression of genes cloned to the plasmids by using of inducer IPTG. Lipoprotein signal sequence is located just ahead of cloning sites of the plasmids, which helps cells to excrete or secrete cloned gene products. Plasmid pUC9 was used as a intermediate vector for transferring of alpha-lFN gene from Hif-2h to pIN vectors in order to solve the problem of different restriction sites between Hif-2h and pIN vectors.

• Archives of Pharmacal Research
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• v.27 no.7
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• pp.776-780
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• 2004

Interferons are cytokines that confer resistance to viral infection and inhibit cellular proliferation. The interferon alpha gene from human blood samples was amplified, cloned and expressed in E. coli (BL21). Leukocyte chromosomal DNA was used as a source of template DNA. Using specific primers, the gene for HulFN$\{alpha}$-2b was amplified and inserted into the E. coli vector, pET21b, by ligation of the HindIII and BamHI linkers of the vector and insert. The insert was further analyzed by PCR, DNA restriction mapping and sequencing, and expressed in a suitable E. coli strain. The production of this important cellular protein in the laboratory has significant applications in production of the recombinant pharmaceutical proteins.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.418.1-418.1
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• 2002

Recombinant interferon alpha is widely used for the treatment of diseases including chronic hepatitis C. However. it has drawbacks such as relatively short serum half-life and rapid clearance like other therapeutic proteins. Using PFGylFltioo which is one of the well-established methods to fulfill the requirements of a long-lasting form of protein. wIe prepared mono-PEG modified interferon alpha-2a in which polyethylene glycol moiety was covalently attached to the amino groups of interfelon alpha-2a. (omitted)

• Journal of Yeungnam Medical Science
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• v.10 no.1
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• pp.190-196
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• 1993

The prognosis of chronic hepatitis C is very variable. In some, the disease is progressive and cirrhosis can develop from chronic hepatitis C. Hepatitis C virus(HCV) may act as a trigger towards hepatocellular carcinoma in patients with cirrhosis. Interferon has been used for the treatment of chronic hepatitis C in abroad, 16 patients with chronic C liver disease were treated with ${\alpha}$-interferon (alfa-2b; "Intron A" Schering Corp. Kenilworth. NJ). All patients were given ${\alpha}$-interferon in subcutaneous doses of 3 million units three times weekly for 1 to 9 months. During therapy, CBC and ALT levels were checked weakly to monthly. After therapy, patients were followed for 1 to 8 months. Among 16 patients treated with ${\alpha}$-interferon, progressive decrease of ALT levels was observed in 14(87.5%). In 11 patients(68.8%), ALT levels fell into the normal range during therapy, and in 9 of 11, within one month after therapy, 6 months after the completion of therapy in 4 of 9 patients(44.4%) whose ALT levels were in the normal range, ${\alpha}$-interferon seems to have effect in controlling disease activity in patients with chronic hepatitis C. But the changes in the usage of ${\alpha}$-interferon, dose and duration, long term follow up and more convenient and simple tests for HCV detection are recommended for the better effect and the exact evaluation on the effect of ${\alpha}$-interferon therapy in patients with chronic hepatitis C.

• KSBB Journal
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• v.5 no.3
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• pp.195-200
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• 1990

The growth behavior of recombinant Escherichia coli cells having plasmid pIF-III-B, which carries human alpha-interferon gene under the control of lpp promoter, lac promoter and lac operator, was studied by using of various E. coli host strains. Expression of the alpha-IFN gene is controllable by using inducer IPTG because the plasmid also contains lacI gene which produces lac regressors. The repressors block the transcription of alpha-IFN gene. There were considerable differences in cell growth according to the host strains used. Cell growth was inhibited not only by plasmid pIF-III-B itself but also by the induction of alph-a-IFN gene expression. Growth inhibition caused by the plasmid itself was more serious than that caused by the induction of alpha-IFN gene expression.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.131-131
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• 1993

위암의 화학요법 성적이 만족스럽지 못하므로 기존의 항암제와 alpha-interferon을 병용사용하므로써 상승적 항암효과가 있는가를 연구하였다. R-25 flask에 위암세포주 SNU-1 및 SNU-16을 배양하면서 항암제 5-FU 혹은 cisplatin에 alpha-interferon 2A(제일제당)를 분자량의 비에 따라 병요처리하였다. 96시간 배양후 cytotoxicity를 Mosman의 방법에 따른 MIT방법으로 분석하였으며, 이를 Chou등의 combination index 분석 program을 처리하였다. 위암세포주 SNU-1에 대하여 5-FU와 alpha-IFN은 상승작용을 보였다. 5-FU의 $IC_{50}$/가 14.25$\mu$M 이었고 IFN처리시는 $IC_{50}$/가 0.02$\mu$M이었으며 combination index(CI)는 모든 용량에서 1이하였다. SNU-16에 대하여도 5-FU와 alpha-IFN의 병용사용은 상승적 작용을 나타내었다. (CI<1.0). cisplatin과 alpha-IFN의 병용시에는 CI가 1이상을 나타내어 상승작용을 증명할 수 없었다.

• Archives of Pharmacal Research
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• v.29 no.5
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• pp.405-411
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• 2006

Biphenyl dimethyl dicarboxylate (DDB) is a hepatoprotectant, which is used as an adjuvant agent in a treatment for chronic hepatitis. Amantadine is an antiviral agent, which is utilized primarily in the treatment of influenza, but also, occasionally in the treatment of hepatitis C. In a previous study, we reported that DDB, coupled with amantadine, would exert an anti-HBV effect, via the induction of interferon-inducible gene expression in the HepG2 2.2.15 cell line. The primary objective of the present study was to determine whether or not DDB and/or amantadine exhibit anti-HBV properties, and what mechanisms of action might be involved in such properties. In our study, we were able to determine that DDB stimulates Jak/Stat signaling, and induces the expression of interferon alpha $(IFN-\alpha)$ stimulated genes, most notably 6-16 and ISG12. In addition, the antiviral effectors induced by $IFN-\alpha$, PKR, OAS, and MxA, were regulated in the presence of DDB at its optimal concentration $(250{\mu}g/mL)$, to a degree commensurate with the degree of induction associated with the $IFN-\alpha$ treated group. Finally, we determined that the replication of pregenomic RNA and HBeAg was inhibited by DDB treatment, and this inhibition was maximized when coupled with the administration of amantadine $(25{\mu}g/mL)$. In conclusion, the results of this study demonstrated clearly that DDB, as well as the combination of DDB/amantadine, directly inhibited $IFN-\alpha$ signaling-mediated replication of HBV in infected hepatocytes, and thus may represent a novel treatment for chronic hepatitis B, which would be characterized principally by its improved safety over other treatment strategies.

• Clinical and Experimental Pediatrics
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• v.63 no.2
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• pp.52-55
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• 2020

Background: Immunomodulatory properties of interferon (IFN) have been documented. It may induce autoimmune diseases such as autoimmune thyroiditis with hypo- or hyperthyroidism. In addition, it may impair thyroid hormone synthesis through affecting iodide organification in thyroid gland. Purpose: The aim of this study was to describe thyroid function tests disturbances in children with chronic hepatitis C (CHC) receiving pegylated interferon-alpha (PEG IFN-α) plus ribavirin. Methods: Fifty children with CHC virus infection who received combined pegylated interferon-alpha with ribavirin were selected. Other 50 apparently healthy children of matched age and sex (considered as control group) were selected. All children (100) were subject to liver function tests, virological studies, and follow-up of thyroid function test during and after the treatment course. Results: Our study showed that 28% of children received combined PEG IFN-α plus ribavirin showed subclinical hypothyroidism. After 24 weeks treatment with combined therapy of IFN plus ribavirin, the mean level of thyroid stimulating hormone (TSH) was 3.23±88 mU/mL, while TSH was 1.16±0.77 mU/mL before starting treatment. On the other hand, mean TSH was 1.09±0.92 mU/mL in normal control group. Conclusion: This study revealed an association between subclinical thyroid dysfunction and treatment with IFN-alpha and ribavirin in children. Further studies on larger number of patients and longer follow-up duration are recommended for further confirmation.

• Archives of Pharmacal Research
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• v.28 no.4
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• pp.451-457
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• 2005

Experimental studies have demonstrated that the triple combination of amantadine (A)/ ursodeoxycholic acid (UDCA, U)/ biphenyl dimethyl dicarboxylate (DDB, D) might have a preferential antiviral effect compared with that observed in interferon-induced antiviral signal pathways, such as those of $STAT1\alpha$ and the 6-16 genes. To confirm the results, this study examined whether th signal transduction for the antiviral activity in HepG2 2.2.15 was induced dependently or independently of interferon. To accomplish this, the correlation between the $STAT1\alpha$ and 6-16 genes, and nitric oxide, for the mediation of the antiviral activity was assessed. The increase in nitric oxide in the UDCA groups suggests that the inhibition of viral gene replication was enhanced by the amantadine combinations (AU and AUD), and might be more effective if incubated for longer periods. It was found that $STAT1\alpha$ was activated by the amantadine combination, although to a lesser extent than that of $interferon-\alpha$, and the primary endpoints examined for the inhibition of gene expression (HBsAg and HBcAg) were remarkably well regulated. This suggests that the amantadine triple, or at least the double, combination had better clinical benefits than those of $IFN-\alpha$ and the nucleoside analogue single treatment. This demonstrates that the amantadine combination might be a substitute for the existing HBV therapy if the results of in vivo and in vitro studies concur.