Showing 3 results for Gholami
Asghar Abdoli, Hoorieh Soleimanjahi, Abbas Jamali, Shima Gholami, Abdolhossein Amini Rissehei, Nabiollah Namvar Asl, Peyvand Biglari, Masoumeh Tavassoti Kheiri,
Volume 1, Issue 1 (Vaccine Research 2014)
Abstract
Introduction: The preparation of seasonal influenza virus vaccines and especially its large-scale production requirement after the emergence or reemergence of a pandemic will need an alternative host cell system due to current suboptimal methods and the insufficiency of embryonated chicken eggs needed for producing them. In response to the vital and increasing demand for alternative means for influenza vaccine production, a cell line culture on microcarriers could be a potential alternative to the egg-based production. Methods: Influenza A/PR/8/1934 H1N1 was purified and quantified by plaque assay. The purified virus with 0.01 multiplicity of infection (MOI) was inoculated on Madin-Darby canine kidney-Siat1 (MDCK-SIAT1) cell line. Cytodex-1 microcarrier beads (2 g/l and 2.0×105cells/ml) were used in a spinner flask to culture MDCK-SIAT1cells .The culture medium was harvested and clarified and the virus yield was quantified by 50% cell culture infective dose ( CCID50) and hemagglutination assays. Next, the virus was concentrated and purified by ultra-filtration and ultra-centrifugation, respectively. Results: MDCK-SIAT1cells attached to the microcarriers and the cell numbers were increased efficiently. The cellular yield from the microcarrier culture was 2×106 cells/ml after 4-5 days. The yield of the virus titer measured by CCID50 and hemagglutination assays after the clarification was 108 CCID50/ml and 40960 HA unit/ml, respectively. Conclusion: MDCK-SIAT1cells may be considered as a new substrate for the production of influenza vaccines. Using Cytodex-1 microcarrier beads can be an appropriate strategy to improve the viral yield and to lower the cost of influenza vaccine production. Vac Res , 2014, 1 (1): 36-40
Tahereh Taheri, Elham Gholami, Faeze Saatchi, Negar Seyed, Yasaman Taslimi, Sima Rafati,
Volume 1, Issue 1 (Vaccine Research 2014)
Abstract
Introduction: Leishmania tarentolae (L. tarentolae) is a nonpathogenic species of Leishmania genus that can be used as an expression system to produce immunogenic proteins or epitopes in vivo acting as an efficient and safe recombinant live vector in vaccinology. Cysteine proteases (CPs) are one group of Family C1 peptidases in Leishmania that are important for survival of the parasite within the host and are introduced as potential vaccine candidates. Two types of cysteine proteases, CPA and CPB, play a critical role in Leishmania. Previously, it has indicated that immunization with two genes or recombinant proteins of CPA and CPB individually or fused together with various adjuvant or combined with liposome are able to elicit a protective immune response against L. major in BALB/c mice. Methods: In this study, for the first time, two lines of recombinant non-pathogenic Leishmania were generated by transfection of a heterologous triple fusion gene encoding cpa/cpb/egfp. For this purpose, two different expressions approaches were taken episomal expression using rDNA promoter, and integrative expression from rRNA locus of genome. Results: After genotype confirmation, the fluorescence intensity was monitored in different phases of parasite growth by both fluorescence microscopy and FACS analysis.Western blot and RT-PCR analysis showed that cpa/cpb/egfp tri-fused genes were specifically expressed in both lines of transgenic parasites. Discussion: In this study a single fused gene was expressed in both systems and the results showed that the expression of integrated gene was higher than episomal. In addition, the present data suggest that L. tarentolae expressing cpa/cpb/egfp is a promising carrier as vectored vaccine in future research. Vac Res , 2014, 1 (1): 1-9
Ashkan Alamdary, Alireza Gholami, Mohammad Azizi, Zahra Noormohammadi,
Volume 8, Issue 1 (6-2021)
Abstract
Introduction: Rabies is almost always fatal but entirely preventable through proper vaccination. Inadequacy of costly high-quality cell culture vaccines is sometimes a bottleneck for expanded rabies control plans. Reverse genetics along with other molecular biology means are trying to improve the immunogenicity and yield of rabies vaccine products. Methods: An additional glycoprotein gene of the rabies virus PV strain was inserted between the glycoprotein and polymerase genes of the virus. The viral proteins were expressed at the T7BHK cell line to rescue the recombinant virus. Results: The recombinant virus containing two consecutive glycoprotein genes was rescued from T7BHK cells. The virus particles were functional and successfully infected the permissive BSR cell line. Conclusion: The new virus strain with an additive copy of the glycoprotein gene has a good potential to be utilized in different studies, including cell biology and immunological properties of the rabies virus. In this study, the recombinant rabies virus was successfully rescued from cell culture which would pave the way for further investigations on this virus.
