Enhanced dendritic cell immunotherapy to treat glioblastoma
Introduction: Glioblastoma (GBM) patients have a poor survival and are inevitably treatment refractory. The power of enhancing the immune system to combat the chemo-radiotherapy resistant tumour, melanoma, affirms the potential for re-directing the immune response against GBM. Furthermore, immunothe...
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| Format: | Thesis (University of Nottingham only) |
| Language: | English |
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2019
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| Online Access: | https://eprints.nottingham.ac.uk/57082/ |
| _version_ | 1848799429991071744 |
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| author | Adhikaree, Jason |
| author_facet | Adhikaree, Jason |
| author_sort | Adhikaree, Jason |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Introduction: Glioblastoma (GBM) patients have a poor survival and are inevitably treatment refractory. The power of enhancing the immune system to combat the chemo-radiotherapy resistant tumour, melanoma, affirms the potential for re-directing the immune response against GBM. Furthermore, immunotherapy combines anti-cancer cytotoxicity with durable memory responses.
Dendritic cells (DC) orchestrate a central role in the adaptive immune response by antigen uptake, presentation and co-stimulation to effector T-cells. However, tumours have evolved mechanisms to evade immune recognition by DC.
Here we focussed on the myeloid CD1c+ (cDC2) subset, the most abundant circulating DC, to assess the feasibility of adoptive transfer in the next generation of DC vaccines. We also study the novel use of p38 mitogen-activated protein kinase inhibitor (p38i) as an adjunct to improve DC dysfunction and combination therapy with immune checkpoint blockade (ICB).
Method: 6-colour flow cytometry of whole blood was used to enumerate and phenotype 4 circulating DC subsets in GBM patients (n=16) and healthy controls (n=16). Following magnetic bead isolation of DC, the cytokines IL-12 and IL-10 were measured post maturation from GBM patients (n=6) and modelled using healthy DC co-cultured with dexamethasone (Dex) and GBM tumour-derived lysate (TDL). A mixed lymphocyte reaction (MLR) measured T-cell stimulatory ability and phenotype. Migration was assessed using a transwell 5M pore membrane towards a CCL21 gradient and antigen presentation measured using an autologous DC, lymphocyte and peptide co-culture expansion system. Peptide-specific responses were analysed by IFN- ELISPOT. cDC2 ability to cross present gp100 peptides was analysed using transfected gp100280-288 T-cell receptor-Jurkat cell line. The p38i, BIRB0796, was added as an adjunct prior to maturation to assess response and ICB added to determine synergy in a MLR.
Results: GBM patients had reduced cDC2 and plasmacytoid DC, with an immature phenotype, compared to controls. Likewise, cDC2 isolated from patients, showed significantly suppressed IL-12 secretion. This was modelled in healthy DC by co-culture with Dex and TDL. Treatment with p38i reversed cDC2 dysfunction with increased co-stimulation expression, IL-12 secretion and allo-stimulation. In addition, IL-10 was suppressed, migration towards CCL21 improved and cross-presentation ability maintained. Patient-derived cDC2 showed improved IL-12:IL-10 ratio, similar to healthy controls. There was a superior and synergistic Th1 proliferation when combined with ICB.
Conclusions: Circulating DC from GBM patients are dysfunctional and this represents a potential mechanism of immune evasion. Here we establish that p38i restores phenotype and function to activate a Th1 response. We have also shown favourable characteristics of a combination ICB treatment strategy. This is the first study in GBM patients to investigate the use of cDC2 in the next generation of vaccines with a p38i adjunct. We are in the process of establishing the first cDC2 vaccine trial in GBM patients. |
| first_indexed | 2025-11-14T20:35:32Z |
| format | Thesis (University of Nottingham only) |
| id | nottingham-57082 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T20:35:32Z |
| publishDate | 2019 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-570822025-02-28T14:36:11Z https://eprints.nottingham.ac.uk/57082/ Enhanced dendritic cell immunotherapy to treat glioblastoma Adhikaree, Jason Introduction: Glioblastoma (GBM) patients have a poor survival and are inevitably treatment refractory. The power of enhancing the immune system to combat the chemo-radiotherapy resistant tumour, melanoma, affirms the potential for re-directing the immune response against GBM. Furthermore, immunotherapy combines anti-cancer cytotoxicity with durable memory responses. Dendritic cells (DC) orchestrate a central role in the adaptive immune response by antigen uptake, presentation and co-stimulation to effector T-cells. However, tumours have evolved mechanisms to evade immune recognition by DC. Here we focussed on the myeloid CD1c+ (cDC2) subset, the most abundant circulating DC, to assess the feasibility of adoptive transfer in the next generation of DC vaccines. We also study the novel use of p38 mitogen-activated protein kinase inhibitor (p38i) as an adjunct to improve DC dysfunction and combination therapy with immune checkpoint blockade (ICB). Method: 6-colour flow cytometry of whole blood was used to enumerate and phenotype 4 circulating DC subsets in GBM patients (n=16) and healthy controls (n=16). Following magnetic bead isolation of DC, the cytokines IL-12 and IL-10 were measured post maturation from GBM patients (n=6) and modelled using healthy DC co-cultured with dexamethasone (Dex) and GBM tumour-derived lysate (TDL). A mixed lymphocyte reaction (MLR) measured T-cell stimulatory ability and phenotype. Migration was assessed using a transwell 5M pore membrane towards a CCL21 gradient and antigen presentation measured using an autologous DC, lymphocyte and peptide co-culture expansion system. Peptide-specific responses were analysed by IFN- ELISPOT. cDC2 ability to cross present gp100 peptides was analysed using transfected gp100280-288 T-cell receptor-Jurkat cell line. The p38i, BIRB0796, was added as an adjunct prior to maturation to assess response and ICB added to determine synergy in a MLR. Results: GBM patients had reduced cDC2 and plasmacytoid DC, with an immature phenotype, compared to controls. Likewise, cDC2 isolated from patients, showed significantly suppressed IL-12 secretion. This was modelled in healthy DC by co-culture with Dex and TDL. Treatment with p38i reversed cDC2 dysfunction with increased co-stimulation expression, IL-12 secretion and allo-stimulation. In addition, IL-10 was suppressed, migration towards CCL21 improved and cross-presentation ability maintained. Patient-derived cDC2 showed improved IL-12:IL-10 ratio, similar to healthy controls. There was a superior and synergistic Th1 proliferation when combined with ICB. Conclusions: Circulating DC from GBM patients are dysfunctional and this represents a potential mechanism of immune evasion. Here we establish that p38i restores phenotype and function to activate a Th1 response. We have also shown favourable characteristics of a combination ICB treatment strategy. This is the first study in GBM patients to investigate the use of cDC2 in the next generation of vaccines with a p38i adjunct. We are in the process of establishing the first cDC2 vaccine trial in GBM patients. 2019-12-11 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/57082/1/Newfinalthesis.pdf Adhikaree, Jason (2019) Enhanced dendritic cell immunotherapy to treat glioblastoma. PhD thesis, University of Nottingham. Dendritic cells; Glioblastoma; Immunotherapy |
| spellingShingle | Dendritic cells; Glioblastoma; Immunotherapy Adhikaree, Jason Enhanced dendritic cell immunotherapy to treat glioblastoma |
| title | Enhanced dendritic cell immunotherapy to treat glioblastoma |
| title_full | Enhanced dendritic cell immunotherapy to treat glioblastoma |
| title_fullStr | Enhanced dendritic cell immunotherapy to treat glioblastoma |
| title_full_unstemmed | Enhanced dendritic cell immunotherapy to treat glioblastoma |
| title_short | Enhanced dendritic cell immunotherapy to treat glioblastoma |
| title_sort | enhanced dendritic cell immunotherapy to treat glioblastoma |
| topic | Dendritic cells; Glioblastoma; Immunotherapy |
| url | https://eprints.nottingham.ac.uk/57082/ |