Identification of T Cell and B Cell Specific Peptide Vaccine for the Treatment of Ross River Virus

: Introduction: Ross River virus, also known as Ross River fever and epidemic polyarthritis, is an infection that is spread to humans through mosquitoes.Ross River virus infection can cause fever, rash, joint inflammation and pain, fatigue and muscle aches. Most people recover completely within 3 to 6 months, although some people have intermittent symptoms for a year or more. Ross River virus infection is caused by an alphavirus(RNA Virus).Ross River virus is named after the Ross River in Townsville, which is the place where it was first identified. Ross River fever is the most common mosquito-borne disease in Australia, and nearly 5000 people are reported to be infected with the virus each year.The T and B lymphocytes (T and B Cells) are involved in the acquired or antigen-specific immune response given that they are the only cells in the organism able to recognize and respond specifically to each antigenic epitope.Developing a B-cell and T-cell specific peptide vaccine for Ross River Virus (RRV) would involve identifying specific antigenic targets that can stimulate the immune response. Materials and methods: Sequence was retrieved, B cell epitope and T cell epitope prediction was carried out and the results were plotted in an excel sheet and graphical representation of immunogenicity regions of Ross Virus was plotted Results: In the B cell epitope prediction ;Peptide RNQPYLFKTNPNYKGNDIKCTSTSRDK was selected as it was homologous (100%) to the C. burnetiid htpB and the length was appropriate for a candidate peptide vaccine. In the T cell epitope prediction ; Conclusion: The discovery and development of B cell and T cell specific peptide vaccines has the potential to provide a personalised and successful strategy to combating Ross River Virus infection, hence providing hope for future prevention and treatment techniques against this viral disease.


Introduction:
Viruses cause a great deal of harm to people and are the primary global cause of morbidity and mortality from infectious diseases.Since they were discovered in antiquity, viral illnesses in humans have influenced • Email: editor@ijfmr.com

IJFMR23069691
Volume 5, Issue 6, November-December 2023 2 human history.The 19th century saw the emergence of scientific methods for the research of virus and infectious disease, which eventually led to the discovery of certain viral disease entities.Many viral infections have been identified and numerous viral diseases have been distinguished (e.g., smallpox vs. chickenpox and measles vs. rubella) thanks to meticulous clinical observations.The pathophysiology of multiple viral diseases has been identified thanks to advancements in our comprehension of disease at the level of cells and tissues, best demonstrated by the groundbreaking research of Virchow.
At the close of the 19th century, the initial viruses were discovered.The tobacco mosaic virus was detected by Ivanovsky and Beijerinck, whereas the foot-and-mouth disease virus was found by Loeffler and Frosch.The U.S. Army Yellow Fever Commission (Reed W et al. 1902)(1)and Walter Reed conducted revolutionary studies on the etiology of yellow fever shortly after these discoveries, leading to the identification of the yellow fever virus.
Australia and the nearby Pacific Islands are home to the endemic viral disease known as Ross River fever, which is spread by mosquitoes.It belongs to the arthritogenic group of alphaviruses, it is a single stranded RNA virus primarily responsible for the disease's crippling polyarthritis, rash, and fever.There is no approved medication or vaccination for this condition, and our understanding of the mechanisms behind human protective humoral immunity is lacking(Powell et al 2020) (4).RRV is transmitted through the bite of Aedes and Culex mosquitoes.Typical infection symptoms include fever, rash, and-most importantlydisabling pain in the muscles and joints that lasts for three to six months (Harley D et al 2001).(5)Since the disease's national alert program was established in 1993, 4,600 cases on average have been recorded in Australia per year for RRV( Australian government department of health).In the past, it was believed that kangaroos and wallabies, which are native to Australia, were the reservoirs of RRV (Claflin et al 2015).(6)Recent data, however, suggests that additional mammalian species, including flying foxes, rabbits, and rodents, may serve as the virus's reservoirs and aid in its dissemination (Lau C et al 2017) (7).This discovery raises questions about the possibility of RRV spreading outside the borders of Australia and the Pacific Territories and raises concerns about the transmission of the virus to humans in the future.
Vaccines are undoubtedly the most successful biological achievement in illness prevention.Over 100 million children worldwide receive vaccines each year to avert diseases that were once common and connected to significant medical issues or even death.Childhood immunizations that are globally provided comprise those for ,mumps , measles,influenza virus which is seasonal ,rubella, hepatitis B ,tetanus, polio,pertussis , diphtheria and others.Vaccines for illnesses that are endemic to specific locations, such as Yellow fever virus, whose mosquito vectors spread year-round in tropical and subtropical climates, are also given to the broader population.Vaccination is predicted to prevent between 2-3 million lives each year (WHO).
Because many vaccines against viral infections are predicated on generating antibody responses, they are often poor stimulants of T cell responses (Plotkin SA et al 2013) (8).T cell stimulating vaccines are needed because T cells are vital in protecting against numerous viral diseases.Specific T cell responses can be elicited by inserting tiny protein fragments (peptides) in a vaccination that can be delivered by MHCmolecules to CD4+ and CD8+ T cells.The primary benefit of peptide vaccines versus traditional immunizations is the ability to specifically trigger T cell responses and the ease with which these vaccines may be produced.Aichele et al. published the initial synthetic peptide vaccination capable of inducing a T cell response in mice.A 15-mer peptide isolated from the NP protein of LCMV was suspended in incomplete Freund's adjuvant (IFA) in this vaccination (Aichele P et al 1990) (9).Further research revealed that such peptide vaccines could provide some protection against virus challenge (Kast WM ate al 1991) (10).These promising results constituted the primary motivation for the current study.
Therefore the current study employed the identification of specific T cell and b cell peptide vaccines against Ross River Virus.

Materials and methods: Sequence retrieval:
All the available Protein databases were evaluated for potential targets for the discovery of vaccination candidates using the server https://www.uniprot.org/proteomes.The NCBI Reference Sequence: (GenBank: NC_075016) structural polyprotein of Ross river virus was chosen for the investigation.The amino acid sequence for the preceding entry was obtained from the NCBI database.Fasta sequences were utilised to predict B and T cell epitopes.

Linear B cell epitope prediction:
Using the BepiPred 2 software application, the protein sequences were utilised to anticipate probable linear B-cell epitopes .As a default, the epitope threshold was set at 0.5.The program's default scoring for epitope (E) is 0.5, and adjustments to this would modify the degree of sensitivity and specificity of the epitope's immunogenic efficacy [13].Positions over the threshold and longer than 20 mer were deemed possible Bcell epitopes.The BepiPred program took into account each epitope's structural predictions (sheet, helix, or coil), epitope locations, and surface accessibility (hidden or exposed).

T cell epitope prediction:
T cell epitope prediction is to find the shortest peptides present in an antigen that can activate T cells or elicit immunogenicity [39].These antigens can activate either CD4+ or CD8+ T lymphocytes.The NetCTL 1.2 server can forecast CTL epitopes.The chosen protein was submitted, and MHC I -binding T cell epitope prediction was performed using the internet server program http://tools.iedb.org/mhci/.The Immune Epitope Database (IEDB), which contains an archive of scientifically identified T cell epitopes, contains information regarding MHC ligand elution and MHC binding studies.Using the HLA allele reference set, the interacting alleles (MHC I-binder) with these epitopes were then identified.

Results: Linear B cell epitope prediction:
The anticipated Linear B cell epitope for Ross River virus varies in length from 3 to 27 mer.The peptide RNQPYLFKTNPNYKGNDIKCTSTSRDK was chosen because it was 100% identical to the C. burnetiid htpB and the length was adequate for a potential peptide vaccine.According to Grifoni et al 2020, the found B-and T-cell epitopes may aid in the creation of powerful peptide-based vaccines to combat the SARS-CoV-2 challenge.Those epitopes without mutations from conserved areas, in particular, may develop immunity that is not only cross-protective across Beta coronaviruses but also reasonably resistant to ongoing virus evolution.(10-13) Lincon et al 2023 proposed that the postulated T cell epitope "ILFLMSQRY" showed increased affinity for interacting with the receptor of its target and would elicit a powerful immune response and operate as a therapeutic agent against monkeypox virus infection in another silico investigation.This highly immunogenic and nonallergenic epitope can also efficiently interact with the human leukocyte antigen HLA-B15:01.
Though no study has been done on identification of B cell and T cell epitopes for Ross River Virus similar studies employed as above had similar results to the present study on the basis of efficacy of T cell and B cell epitope vaccine against the treatment of a virus

Limitation of the study:
Extensive research and efficacy can be obtained by conducting the study in Vivo .

Conclusion:
A B cell peptide vaccination can induce the development of antibodies designed to recognize and neutralize the virus by targeting specific surface proteins or epitopes unique to RRV.This method can keep RRV from infecting host cells and spreading throughout the body.T cells play an important part in the immune response by attacking infected cells directly and coordinating the overall immune defence.A T cell-specific peptide vaccination can activate these cells to recognize and kill RRV-infected cells, lowering viral replication and infection severity.Furthermore, using both B cell and T cell peptide vaccines in a dual approach for comprehensive immune response can provide a comprehensive resistance against RRV.

Table 1 . Prediction of B cell epitope T cell epitope prediction :
The IEDB tool peptide_binding_to_MHC_class_I_molecules and the HLA class I set(Weiskopf et al.,  2013)were used to determine T-cell epitopes for the protein.Percentile rank with the least threshold was set as the best peptide score( threshold ranges from 0 to 1%) resulting in a high antigenic score.A peptide with a high antigenicity score as well as the ability to bind to a greater number of alleles is thought to have high potential to elicit a powerful defence response.The highest potential peptides were selected as ATNPSVFV with a score of 0.989845 and percentile rank or antigenicity score of 0.01% and the second highest being ETAIGNFF with score of 0.973605 and percentile rank of 0.01.Protein-peptide interactions play an important role in biological signaling networks.The protein's peptides 'ATNPSVF' and 'ETAIGNFF' were predicted to bind to HLA-A*11:01 and HLA-A*68:01.There were 31 reported epitopeprotein interactions, and the best prediction was chosen.vaccine design since it can generate long-lasting protection.Predicting the antigenicity of all protein sequences is a key part of vaccine development since only antigenic peptides can elicit an immune response in the host.