Int. J Sci. Emerging Tech Vol-4 No 3 September 2012 153 Mutations in ISDR may be linked to High VIREMIA and Virus Resistance to IFN-Alpha-2b But Responsive to PEG-IFN-Alpha-2a Abida Raza 1 *, Nasib Zaman 2 , Binish Gull Arshad 3 , Muhammad Javed Asad 2 , Hafsa Aziz 1 , Samina N. Qureshi 3 , Javaid Irfan 1 1 Postdoc School of Pharmacy, University of London, Senior Scientist, Nuclear Medicine, Oncology and Radiotherapy Institute (NORI), Islamabad, Pakistan Email: abida_rao@yahoo.com 2 PMAS, Arid Agriculture University, Rawalpindi 3 Quaid-i-Azam University, Islamabad Abstract: Mutation in interferon sensitivity determining region may play role in virus resistance. Genotype 3a patient was subjected to 48 weeks combination therapy of IFN-alpha-2b plus ribavirin but he showed high levels of viremia before, during and after treatment although his ALT level became normal. His IL-8 and TNF-alpha levels werefound quite high before and after IFN-alpha-2 combination therapy. While comparing its ISDR-NS5A with end of treatment responder patient, eight mutations were observed in a 52 amino acid protein residue. Patient was advised for PEG-IFN- alpha-2a combination therapy for 24 weeks, to which he responded well after 4 week and showed sustained virologic response after 06 months of completion of therapy. His IL-8 and TNF-alpha levels also came to lower levels after treatment with PEG-IFN-alpha 2a combination therapy. In phylogenetic tree its genome (NZ1) along with another nonresponder case (NZ2) was placed close to Brazilian isolates. NZI and NZ2 showed 87 % sequence homology with each other while NZ1 had 89% sequence homology with EF208017 and 87% with EF20995. NZ2 showed 91% homology with EF208017 and 98% with EF207995 which is quite interesting. Mutations in ISDR sequence may be the reason for non response to IFN combination therapy of this HCV genotype 3 patient. ISDR of genotype 3 along with IL-8 and TNF-alpha may be screened on larger scale in Pakistani population which may help in deciding a cost effective treatment plan. Key words: NS5A-ISDR mutations; IFN-alpha-2b; PEG- IFN; HCV; IL-8; Pakistan 1. Introduction Mutations in interferon sensitivity determining region (ISDR) has been linked to Hepatitis C virus resistance in some studies from Japan in genotype 1 [1]-[3] but contradictory reports have been submitted from other parts of the world including Pakistan [4]-[8]. If three or more than three mutations are observed in ISDR region, it is declared as mutant ISDR strain, while intermediate mutant type has one to three amino acid mutations 4. The number of amino acid substitutions in ISDR in the non-structural 5A (NS5A) gene of hepatitis C virus (HCV) is closely associated with the interferon (IFN) response and viral load [9]. Mutations in ISDR may play role in virus replication and in some cases increase the replication many folds. Most of the data available is on viral genotype 1b. In Pakistan most common genotype is 3a and fortunately above 70 percent response to 24 week treatment plan of non pegylated interferon (non PEG-IFN) combination therapy has been observed [10]. Pegylated interferon combination therapy (PEG-IFN) being quite expensive is not affordable so usually it is not first choice for poor patients. There is need to individualize the treatment plan. For the current case we have tried to explore the NS5A-ISDR amino acid sequence with viral load, virus genotype, ALT, IL-8 protein, TNF-alpha levels with response to interferon therapy with change in treatment plan. 2. Methods Study involves the human subjects; it was approved by the ethical review committee of the Institute. Moreover written consent forms dually signed by the patients were also taken before the start of the study. 3. Patient Clinical Details A 50-year-old male patient body mass index 20.8 belonging to district Mandibahauddin in upper Punjab, Pakistan was diagnosed Hepatitis C in 1995. He was admitted with general fatigue, high-grade fever, and liver dysfunction. No evidence of prior liver disease was found, and the patient had no history of drug or alcohol consumption. The infection route of HCV was obscure. In the last 6 months, he had not received any blood transfusions, taken any drugs intravenously, undergone acupuncture, nor had sexual contact with a known hepatitis virus carrier __________________________________________________________________________ International Journal of Science & Emerging Technologies IJSET, E-ISSN: 2048 - 8688 Copyright © ExcelingTech, Pub, UK (http://excelingtech.co.uk/)