Part:BBa_K3504020

Multi-Epitope TNBC Vaccine Version (2)

NOTICE: Parts in our range for this season have been created as a part of our Phase I design of our project. These parts HAVE NOT been tested or characterized in the lab due to COVID-19-related precautionary measures. We have enriched our new parts pages with data from literature and results from our modeling and simulations. If you are intending on using this part or others in our range, please keep in mind these limitations and update these parts with data from your experimentation. Feel free to reach us at: igem.afcm@gmail.com for further inquiries.

Part Description

A multi-epitope vaccine formed of highly expressed and specific TNBC neo-epitopes and specifically chosen according to egyptian population alleles which can work as a generalized vaccine and also personalized vaccine which would illicit an immune response specific to TNBC tumor cells

Usage

Immune-modulating adjuvants and PADRE (Pan HLA-DR epitopes) sequence were added with epitopes sequence to enhance the immunogenicity. All the epitopes, adjuvants and PADRE sequence were joined by linkers.

Through the help of the EAAAK linker at the start (to the MEV N-terminal) the adjuvant (45 amino acid long β-defensin) was bound. EAAK linker is found to increase stability and reduces connection with other protein areas with efficient detachment. There is a possibility that the immunogenicity could increase with an adjuvant. Based on the interactions’ compatibility epitopes were merged together sequentially with AAY and GPGPG linkers, respectively. In the construction of multiepitope vaccines AAY and GPGPG have a main task to prevent the production of junctional epitopes and also promote the immunization and the presentation of epitopes. CTL-epitopes were coupled with AAY linkers and HTL epitopes were coupled with GPGPG and that permitted proficient dissociation and epitope identification.

A natural link was established among both innate and adaptive immune responses through Heat-shock proteins(hsp) by merging the idyllic properties of antigen carriage (chaperoning), targeting and activation of antigen-presenting cells (APC), including dendritic cells (DC). The uptake of hsp complexes by DC allows proficient capture and presentation of pathogen-specific antigens and also permits the mounting of a specific immune response by the production of CD4+ T-cell responses.

To improve the vaccine efficacy and potency non-natural pan DR (PADRE) 13 amino acid epitope (AKFVAAWTLKAAA) that induce CD4+ T-cells were also combined along with the adjuvants. Heat Shock Protein (hsp) was retrieved from the database to design a multi-epitope subunit vaccine containing a CTL, HTL and BCL epitopes of varying length.

Characterization

Figure(1): Using C-IMMSIM simulation models to describe both Humoral and Cellular response of our predicted epitopes in our 1st proposed multi-epitope DNA vaccine for TNBC, Showing Antigen & Immunoglobulins level, B-cell Populations with various isotypes as well as Memory & Not Memory B-Cells and CD4 T-Cell Population & the regulatory T-Cells.

Figure(2): Using C-IMMSIM simulation models to describe both Humoral and Cellular response of our predicted epitopes in our 1st proposed multi-epitope DNA vaccine for TNBC,Showing the CD8 T-Cell Population and NKCs & DCs & MQs & Epithelial Populations.

References

1-Tahir ul Qamar, Muhammad, et al. “Multiepitope-Based Subunit Vaccine Design and Evaluation against Respiratory Syncytial Virus Using Reverse Vaccinology Approach.” Vaccines, vol. 8, no. 2, 1 June 2020, p. 288, www.mdpi.com/2076-393X/8/2/288/htm, 10.3390/vaccines8020288. Accessed 22 Oct. 2020.

2-McNulty, Shaun, et al. “Heat-Shock Proteins as Dendritic Cell-Targeting Vaccines - Getting Warmer.” Immunology, vol. 139, no. 4, 2 July 2013, pp. 407–415, 10.1111/imm.12104. Accessed 18 Nov. 2019.

3-Solanki, Vandana, et al. “Prioritization of Potential Vaccine Targets Using Comparative Proteomics and Designing of the Chimeric Multi-Epitope Vaccine against Pseudomonas Aeruginosa.” Scientific Reports, vol. 9, no. 1, 27 Mar. 2019, 10.1038/s41598-019-41496-4. Accessed 22 May 2020.

4-Castiglione, F., & Bernaschi, M. (2004, April 30). C-ImmSim∗ : Playing with the immune response. Retrieved October 26, 2020, from https://www.math.ucsd.edu/~helton/MTNSHISTORY/CONTENTS/2004LEUVEN/CDROM/papers/316.pdf

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 874
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 208
  Illegal BsaI.rc site found at 694
  Illegal SapI.rc site found at 79