Paper Example on the Role of Interferon Response in Containing Human Pathogenic Bourbon Virus

Introduction

According to recent reports from the southern part of the United States and Midwest have linked the discovery of tick-transmitted viral pathogen known as Bourbon Virus (BRBV) with severe pyretic disease (Kosoy et al, 2015). The disease is a threat to the public health system of the United States. "Thogotovirus" is virus-like influenza with a segmented Ribonucleic acid genome. It is normally found in continents like Africa, Europe, and Asia, and usually connected with ailments of livestock.

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Materials and Methods

Biosafety and Animal Ethics

All the work which involved infectious virus was done under "Biosafety Level 3 situations" due to strange health risk linked with BRBV. The animals were handled in accord with guiding principles of the Federation or Laboratory Animal Science Association and National Welfare Body of Germany. The tests on animals were done accordingly and strictly following the protection laws of the nation.

Cells and Viruses

The cells that were used included Human Lung epithelial cells, Human Hepatoma, (Huh7) Cervix carcinoma, and African Green Monkey Vero cells. The Huh7 cells were used with HIFN-a2a. When assessing the cytotoxicity of these elements the uninfected cells were treated for two days. Afterward, we evaluated the productivity of the cells by calculating the activity of lactate dehydrogenase in the cell culture supernatant.

Viruses and Infection

Virus stock of BRBV, THOV, and DHOV were produced on Vero cells (Kosoy et al, 2015). These cells were infected with BRBV to conduct an experiment of growth kinetics. Virus having supernatants were harvested and stored at -80C.

Animal Infections

Mice which had deformities were bred in the IFN pathway. All tests were done on the mice (7-9 week old). They were infected with BRBV dissolved in 100L PBS with 0.3% bovine serum albumin (Lambert et al, 2015). The weight was calculated, clinical score and survival rates of each animal as well. It was arrived at if the weight loss was less than 25% it means that the mice shows severe medical signs. The organs were obtained (liver, spleen, etc) on the fourth day and they were homogenized through "FastPrep Homogenizer" (Lambert et al, 2015). They were later assessed after the centrifugation of the supernatants using "Plaque assay" on Vero cells. The entire blood from the mice was numb using Ketamine before the cervical disruption. The serum samples were developed at 37C for 10 min and they were used to identify alanine transaminases through color endpoint assay.

Real-Time Reverse Transcription PCR

Ribonucleic acid was removed from the infected cells and subjected to cDNA synthesis. Real-time reverse transcription was done via 10 ng cDNA IN SYBR Green Analyzer.

Results

According to the outcomes, there are fewer people who have austere BRBV infection and they might have gotten the virus as a result of transient weakness in their inner antiviral immune reaction (Tsubota et al, 2002). The study also discovers that potential medical option hell-bent on lengthy approved medication with ribavirin combined with IFN- a future evaluation of "seroprevalence" to BRBV in humans is quickly need to examine the zoonotic spread of the virus in the affected section. Infected laboratory mice did not exhibit any medical signs, nevertheless, the mice having deformities in type 1 and 2 interferon system, BRBV grew to high titers and caused severe pyretic disease. During the cell culture test, BRBV was blocked by antiviral agents like" Ribavirin and favipiravir "

References

Kosoy, O. I., Lambert, A. J., Hawkinson, D. J., Pastula, D. M., Goldsmith, C. S., Hunt, D. C., & Staples, J. E. (2015). Novel thogotovirus associated with febrile illness and death, United States, 2014. Emerging infectious diseases, 21(5), 760. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4412252/

Lambert, A. J., Velez, J. O., Brault, A. C., Calvert, A. E., Bell-Sakyi, L., Bosco-Lauth, A. M., ... & Kosoy, O. I. (2015). Molecular, serological and in vitro culture-based characterization of Bourbon virus, a newly described human pathogen of the genus Thogotovirus. Journal of Clinical Virology, 73, 127-132. Retrieved from https://www.sciencedirect.com/science/article/pii/S1386653215007167

Tsubota, A., Akuta, N., Suzuki, F., Suzuki, Y., Someya, T., Kobayashi, M., ... & Kumada, H. (2002). Viral dynamics and pharmacokinetics in combined interferon alfa-2b and ribavirin therapy for patients infected with hepatitis C virus of genotype 1b and high pretreatment viral load. Intervirology, 45(1), 33-42