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Identification of African swine fever virus proteins that activate T-cell immune responses
Dissertation (MSc (Veterinary Sciences))--University of Pretoria, 2018.
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| Format: | Thesis |
| Language: | English |
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University of Pretoria
2025
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| _version_ | 1867613672802615296 |
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| access_status_str | Open Access |
| author2 | Van Kleef, Mirinda |
| author_browse | Van Kleef, Mirinda |
| author_facet | Van Kleef, Mirinda |
| collection | Thesis |
| dc_rights_str_mv | © 2024 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
| description | Dissertation (MSc (Veterinary Sciences))--University of Pretoria, 2018. |
| format | Thesis |
| id | oai:repository.up.ac.za:2263/107114 |
| institution | University of Pretoria (South Africa) |
| language | English |
| last_indexed | 2026-06-10T12:39:52.674Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | University of Pretoria |
| publisherStr | University of Pretoria |
| record_format | dspace |
| source_str | UPSpace — University of Pretoria Institutional Repository |
| spelling | oai:repository.up.ac.za:2263/107114 Identification of African swine fever virus proteins that activate T-cell immune responses Van Kleef, Mirinda fr33ic3@gmail.com Pretorius, Alri Van Heerden, Juanita Mabetlela, Freddy Mokadi UCTD Sustainable Development Goals (SDGs) Protein Fever Swine Immune T-cell Dissertation (MSc (Veterinary Sciences))--University of Pretoria, 2018. African swine fever (ASF) is a highly contagious haemorrhagic disease of domestic pigs, which affects wild boar as well and for which there is currently no commercial vaccine. African swine fever virus (ASFV), the causal agent of ASF infects warthogs (Phacochoerus africanus), bush pigs (Potamochoerus porcus), giant forest hogs and members of the soft tick genus (Ornithodoros moubata complex) without causing disease. Virulent isolates of this virus causes mortality in domestic pigs within seven to ten days post infection, leading to negative economic consequences. Vaccination attempts using different strategies only protect against homologous strains with inadequate protection against heterologous strains of ASFV. The immunological characterisation of ASFV proteins could assist in the development of an effective vaccine against ASF. The aim of this study was therefore to immunologically characterise a selected range of ASFV proteins (p22, CD2V, pp220 fragments (F)1-F4, pS273R, pA104R, pE165R, pF334L, pK205R and pL11L) by identifing the cytokines they induce in peripheral blood mononuclear cells (PBMC) from a pig that was infected with ASFV via tick feeding (pig 1) and an incontact ASFV infected pig (pig 2). The benefits of identifying immunilogical characteristics of the cytokines can lead to possible vaccine development and decrease of animal losses due to ASF. Several proteins (p22, pp220 F1-4, pS273R, pA104R, pE165R and pK205R) from the genotype II ASFV isolate MAL/11/02 were successfully expressed using a pET102/D-Topo isomerase enzyme (TOPO)® bacterial expression system. Plasmids with gene inserts were amplified using the Invitrogen E. coli TOP10 cloning cells and proteins were expressed in BL21 DE3 cells using isopropyl β-D-1-thiogalactopyranoside (IPTG) induction. The protein product of genes EP402R (CD2V), pF334L and pL11L did not express and were excluded from futher experiments. The expressed proteins were purified by affinity chromatography using Nickel columns with affinity to the His-tag on the recombinant proteins. The immunological assays, interferon gamma (IFN)-γ enzyme-linked immunospot (ELISpot) and cytokine [Interleukin (IL)-2, IL-4, IL-8, IL 10, IL-12, IFN-γ and IFN-alpha (α)] reverse transcription (RT) quantitative real time polymerase chain reaction (RT-qPCR), were conducted using PBMCs. PBMC was collected from pig 1 after ten days post infection (pi) with the genotype XIX ASFV isolate RSA/12/15 strain of ASFV and pig 2 ten days after co-habitation with pig 1. Only four of the nine recombinant ASFV proteins produced significant cytokine levels. The recombinant p22 protein up-regulated cytokines IL-2, IL-4, IL-8, IL-10 and IFN-γ in pig 1 and IFN-α in pig 2. Fragment four (F4) of polyprotein (pp) 220 up-regulated IL-4, IL-8, IL-12 and IFN-α in pig 1 and IFN-α in pig 2. Recombinant proteins pA104R up-regulated IFN-α in pig 1 and pS273R significantly down-regulated IFN-α in pig 1. In conclusion, the study revealed that ASFV recombinant proteins p22 and pp220-F4 induced all the study cytokines which are important in antiviral immunity. This data can be applied in future ASFV vaccinology studies. Veterinary Tropical Diseases MSc (Veterinary Sciences) Unrestricted Faculty of Veterinary Science SDG-03: Good health and well-being 2025-12-08T07:13:32Z 2025-12-08T07:13:32Z 2019-04 2018-09 Dissertation * A2019 http://hdl.handle.net/2263/107114 en © 2024 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria |
| spellingShingle | UCTD Sustainable Development Goals (SDGs) Protein Fever Swine Immune T-cell Identification of African swine fever virus proteins that activate T-cell immune responses |
| title | Identification of African swine fever virus proteins that activate T-cell immune responses |
| title_full | Identification of African swine fever virus proteins that activate T-cell immune responses |
| title_fullStr | Identification of African swine fever virus proteins that activate T-cell immune responses |
| title_full_unstemmed | Identification of African swine fever virus proteins that activate T-cell immune responses |
| title_short | Identification of African swine fever virus proteins that activate T-cell immune responses |
| title_sort | identification of african swine fever virus proteins that activate t cell immune responses |
| topic | UCTD Sustainable Development Goals (SDGs) Protein Fever Swine Immune T-cell |
| url | http://hdl.handle.net/2263/107114 |