In Vitro Priming of Human T Cells by Dendritic Cells Provides a Screening Tool for Candidate Vaccines for <i>Burkholderia pseudomallei</i>
Murine dendritic cells, when pulsed with heat-killed <i>Burkholderia pseudomallei</i> and used to immunise naïve mice, have previously been shown to induce protective immunity <i>in vivo</i>. We have now demonstrated the <i>in vitro</i> priming of naïve human T ce...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-08-01
|
| Series: | Vaccines |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-393X/9/8/929 |
| _version_ | 1797521903574843392 |
|---|---|
| author | Durga Reddi Lydia Durant David Bernardo Alistair Noble Nicholas R. English Philip Hendy Graeme C. Clark Joann L. Prior Ethel Diane Williamson Stella C. Knight |
| author_facet | Durga Reddi Lydia Durant David Bernardo Alistair Noble Nicholas R. English Philip Hendy Graeme C. Clark Joann L. Prior Ethel Diane Williamson Stella C. Knight |
| author_sort | Durga Reddi |
| collection | DOAJ |
| description | Murine dendritic cells, when pulsed with heat-killed <i>Burkholderia pseudomallei</i> and used to immunise naïve mice, have previously been shown to induce protective immunity <i>in vivo</i>. We have now demonstrated the <i>in vitro</i> priming of naïve human T cells against heat-killed <i>B. pseudomallei</i>, by co-culture with syngeneic <i>B. pseudomallei-</i>pulsed dendritic cells. Additionally, we have enriched the DC fraction such that a study of the differential response induced by pulsed DCs of either myeloid or plasmacytoid lineage in syngeneic human T cells was achievable. Whilst both mDCs and pDCs were activated by pulsing, the mDCs contributed the major response to <i>B. pseudomallei</i> with the expression of the migration marker CCR7 and a significantly greater secretion of the proinflammatory TNFα and IL1β. When these DC factions were combined and used to prime syngeneic T cells, a significant proliferation was observed in the CD4<sup>+</sup> fraction. Here, we have achieved human T cell priming <i>in vitro</i> with unadjuvanted <i>B. pseudomallei</i>, the causative organism of melioidosis, for which there is currently no approved vaccine. We propose that the approach we have taken could be used to screen for the human cellular response to candidate vaccines and formulations, in order to enhance the cell-mediated immunity required to protect against this intracellular pathogen and potentially more broadly against other, difficult-to-treat intracellular pathogens. To date, the polysaccharide capsule of <i>B. pseudomallei</i>, fused to a standard carrier protein, e.g., Crm, looks a likely vaccine candidate. Dendritic cells (DCs), providing, as they do, the first line of defence to infection, process and present microbial products to the immune system to direct downstream immune responses. Here, we have sought to use DCs <i>ex vivo</i> to identify immunogenic products from heat-killed <i>B. pseudomallei</i>. Using practical volumes of fresh human donor blood, we show that heat-killed <i>B. pseudomallei</i> activated and stimulated the expression of pro-inflammatory cytokines TNF-α, IL-1β and IL-6 from both myeloid and plasmacytoid DCs. Furthermore, <i>B. pseudomallei-</i>pulsed DCs cultured with naïve syngeneic T cells <i>ex vivo</i>, induced the activation and proliferation of the CD4<sup>+</sup> T-cell population, which was identified by cell surface marker staining using flow cytometry. Thus, both DC subsets are important for driving primary T helper cell responses to <i>B. pseudomallei</i> in healthy individuals and have the potential to be used to identify immunogenic components of <i>B. pseudomallei</i> for future therapies and vaccines. |
| first_indexed | 2024-03-10T08:19:07Z |
| format | Article |
| id | doaj.art-fbc3e589977e4853b2d0d47524bd1be1 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-10T08:19:07Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-fbc3e589977e4853b2d0d47524bd1be12023-11-22T10:08:14ZengMDPI AGVaccines2076-393X2021-08-019892910.3390/vaccines9080929In Vitro Priming of Human T Cells by Dendritic Cells Provides a Screening Tool for Candidate Vaccines for <i>Burkholderia pseudomallei</i>Durga Reddi0Lydia Durant1David Bernardo2Alistair Noble3Nicholas R. English4Philip Hendy5Graeme C. Clark6Joann L. Prior7Ethel Diane Williamson8Stella C. Knight9Antigen Presentation Research Group, Imperial Centre for Translational and Experimental Medicine, 72 Du Cane Road, London W12 0NN, UKAntigen Presentation Research Group, Imperial Centre for Translational and Experimental Medicine, 72 Du Cane Road, London W12 0NN, UKAntigen Presentation Research Group, Imperial Centre for Translational and Experimental Medicine, 72 Du Cane Road, London W12 0NN, UKAntigen Presentation Research Group, Imperial Centre for Translational and Experimental Medicine, 72 Du Cane Road, London W12 0NN, UKAntigen Presentation Research Group, Imperial Centre for Translational and Experimental Medicine, 72 Du Cane Road, London W12 0NN, UKAntigen Presentation Research Group, Imperial Centre for Translational and Experimental Medicine, 72 Du Cane Road, London W12 0NN, UKDefence Science and Technology Laboratory, Porton Down SP4 0JQ, UKDefence Science and Technology Laboratory, Porton Down SP4 0JQ, UKDefence Science and Technology Laboratory, Porton Down SP4 0JQ, UKAntigen Presentation Research Group, Imperial Centre for Translational and Experimental Medicine, 72 Du Cane Road, London W12 0NN, UKMurine dendritic cells, when pulsed with heat-killed <i>Burkholderia pseudomallei</i> and used to immunise naïve mice, have previously been shown to induce protective immunity <i>in vivo</i>. We have now demonstrated the <i>in vitro</i> priming of naïve human T cells against heat-killed <i>B. pseudomallei</i>, by co-culture with syngeneic <i>B. pseudomallei-</i>pulsed dendritic cells. Additionally, we have enriched the DC fraction such that a study of the differential response induced by pulsed DCs of either myeloid or plasmacytoid lineage in syngeneic human T cells was achievable. Whilst both mDCs and pDCs were activated by pulsing, the mDCs contributed the major response to <i>B. pseudomallei</i> with the expression of the migration marker CCR7 and a significantly greater secretion of the proinflammatory TNFα and IL1β. When these DC factions were combined and used to prime syngeneic T cells, a significant proliferation was observed in the CD4<sup>+</sup> fraction. Here, we have achieved human T cell priming <i>in vitro</i> with unadjuvanted <i>B. pseudomallei</i>, the causative organism of melioidosis, for which there is currently no approved vaccine. We propose that the approach we have taken could be used to screen for the human cellular response to candidate vaccines and formulations, in order to enhance the cell-mediated immunity required to protect against this intracellular pathogen and potentially more broadly against other, difficult-to-treat intracellular pathogens. To date, the polysaccharide capsule of <i>B. pseudomallei</i>, fused to a standard carrier protein, e.g., Crm, looks a likely vaccine candidate. Dendritic cells (DCs), providing, as they do, the first line of defence to infection, process and present microbial products to the immune system to direct downstream immune responses. Here, we have sought to use DCs <i>ex vivo</i> to identify immunogenic products from heat-killed <i>B. pseudomallei</i>. Using practical volumes of fresh human donor blood, we show that heat-killed <i>B. pseudomallei</i> activated and stimulated the expression of pro-inflammatory cytokines TNF-α, IL-1β and IL-6 from both myeloid and plasmacytoid DCs. Furthermore, <i>B. pseudomallei-</i>pulsed DCs cultured with naïve syngeneic T cells <i>ex vivo</i>, induced the activation and proliferation of the CD4<sup>+</sup> T-cell population, which was identified by cell surface marker staining using flow cytometry. Thus, both DC subsets are important for driving primary T helper cell responses to <i>B. pseudomallei</i> in healthy individuals and have the potential to be used to identify immunogenic components of <i>B. pseudomallei</i> for future therapies and vaccines.https://www.mdpi.com/2076-393X/9/8/929dendritic cells<i>Burkholderia pseudomallei</i>T cell priming |
| spellingShingle | Durga Reddi Lydia Durant David Bernardo Alistair Noble Nicholas R. English Philip Hendy Graeme C. Clark Joann L. Prior Ethel Diane Williamson Stella C. Knight In Vitro Priming of Human T Cells by Dendritic Cells Provides a Screening Tool for Candidate Vaccines for <i>Burkholderia pseudomallei</i> Vaccines dendritic cells <i>Burkholderia pseudomallei</i> T cell priming |
| title | In Vitro Priming of Human T Cells by Dendritic Cells Provides a Screening Tool for Candidate Vaccines for <i>Burkholderia pseudomallei</i> |
| title_full | In Vitro Priming of Human T Cells by Dendritic Cells Provides a Screening Tool for Candidate Vaccines for <i>Burkholderia pseudomallei</i> |
| title_fullStr | In Vitro Priming of Human T Cells by Dendritic Cells Provides a Screening Tool for Candidate Vaccines for <i>Burkholderia pseudomallei</i> |
| title_full_unstemmed | In Vitro Priming of Human T Cells by Dendritic Cells Provides a Screening Tool for Candidate Vaccines for <i>Burkholderia pseudomallei</i> |
| title_short | In Vitro Priming of Human T Cells by Dendritic Cells Provides a Screening Tool for Candidate Vaccines for <i>Burkholderia pseudomallei</i> |
| title_sort | in vitro priming of human t cells by dendritic cells provides a screening tool for candidate vaccines for i burkholderia pseudomallei i |
| topic | dendritic cells <i>Burkholderia pseudomallei</i> T cell priming |
| url | https://www.mdpi.com/2076-393X/9/8/929 |
| work_keys_str_mv | AT durgareddi invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT lydiadurant invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT davidbernardo invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT alistairnoble invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT nicholasrenglish invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT philiphendy invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT graemecclark invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT joannlprior invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT etheldianewilliamson invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii AT stellacknight invitroprimingofhumantcellsbydendriticcellsprovidesascreeningtoolforcandidatevaccinesforiburkholderiapseudomalleii |