Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1 Xhoi mutation in nasopharyngeal carcinoma
Regulatory T-cells (Tregs) are thought to be involved in the immunopathogenesis of many cancers including nasopharyngeal carcinoma (NPC) which is known to be strongly associated with Epstein-Barr virus (EBV) infection. Loss of EBV latent membrane protein 1 (LMP1) XhoI restriction site has been su...
| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2023
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| Online Access: | http://eprints.usm.my/58895/1/AHMAD%20ADEBAYO%20IREKEOLA-24%20pages.pdf |
| _version_ | 1797014162719637504 |
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| author | Irekeola, Ahmad Adebayo |
| author_facet | Irekeola, Ahmad Adebayo |
| author_sort | Irekeola, Ahmad Adebayo |
| collection | USM |
| description | Regulatory T-cells (Tregs) are thought to be involved in the
immunopathogenesis of many cancers including nasopharyngeal carcinoma (NPC)
which is known to be strongly associated with Epstein-Barr virus (EBV) infection. Loss
of EBV latent membrane protein 1 (LMP1) XhoI restriction site has been suggested to
define EBV strain associated with increased tumourigenicity. Despite the perceived
roles of Tregs and EBV LMP1 XhoI mutation in cancers, their roles in the pathogenesis
of NPC are yet to be fully elucidated. Hence, this study aims to elucidate the proportion
of Tregs, associated cytokines, and LMP1 XhoI mutation in NPC in order to provide
baseline and additional data to foster the strides of current and future immunotherapeutic
approaches. Blood samples were collected from NPC patients (n=23) and healthy
controls (n=23). Tissue biopsy was available for 7 of the NPC patients. Samples were
assessed for Tregs (CD4+FoxP3+, CD4+CD25+FoxP3+, CD8+FoxP3+, and
CD8+CD25+FoxP3+) using multicolour flow cytometry, and XhoI mutation using
polymerase chain reaction. Tregs proliferation was determined by Ki67 expression, and
ex vivo proliferation was assessed after culturing PBMCs with IL-2 and anti-CD3/CD28
mAbs. The expression levels of five cytokines (IL-2, IL-10, IL-6, TNF-α, and IFN-ƴ)
were evaluated using multiplex bead analysis. Tregs were generally found to be elevated in NPC patients. XhoI mutation was detected in both patients (39%) and controls (30%).
The correlation between clinicopathological features and the frequency of circulating
Tregs in the patients revealed no significant correlation for both CD4+ and CD8+ Tregs,
even though Tregs levels were fairly higher in patients with stage IV compared with
stage II/III cancer. In comparison with unstimulated cells, both CD4+ and CD8+ Tregs
proportion heightened following the ex vivo culture of PBMCs. CD4+ and CD8+ Tregs in
the PBMCs obtained from healthy controls responded better to ex vivo stimulation
compared with those from patients. Of the cytokines, significant expression levels were
observed for plasma IL-10 and TNF-α in patients compared with controls. The
concentration of IL-10 was significantly lower in patients (10.47 ± 16.17 versus 16.12 ±
33.38, p = 0.0157). Meanwhile, plasma levels of TNF-α were higher in patients (p
<0.0001) and were found to correlate with CD4+ Tregs in controls but not in patients.
Correlation between IL-2 and CD8+CD25+FoxP3+ (r = -0.4670, p = 0.0438) Tregs was
also observed. Data from this study demonstrates that targeting or developing therapies
that specifically modulate Tregs number, trafficking, or activity, will be promising in
NPC treatment. |
| first_indexed | 2024-03-06T16:12:18Z |
| format | Thesis |
| id | usm.eprints-58895 |
| institution | Universiti Sains Malaysia |
| language | English |
| last_indexed | 2024-03-06T16:12:18Z |
| publishDate | 2023 |
| record_format | dspace |
| spelling | usm.eprints-588952023-08-06T04:08:10Z http://eprints.usm.my/58895/ Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1 Xhoi mutation in nasopharyngeal carcinoma Irekeola, Ahmad Adebayo QH573-671 Cytology R Medicine Regulatory T-cells (Tregs) are thought to be involved in the immunopathogenesis of many cancers including nasopharyngeal carcinoma (NPC) which is known to be strongly associated with Epstein-Barr virus (EBV) infection. Loss of EBV latent membrane protein 1 (LMP1) XhoI restriction site has been suggested to define EBV strain associated with increased tumourigenicity. Despite the perceived roles of Tregs and EBV LMP1 XhoI mutation in cancers, their roles in the pathogenesis of NPC are yet to be fully elucidated. Hence, this study aims to elucidate the proportion of Tregs, associated cytokines, and LMP1 XhoI mutation in NPC in order to provide baseline and additional data to foster the strides of current and future immunotherapeutic approaches. Blood samples were collected from NPC patients (n=23) and healthy controls (n=23). Tissue biopsy was available for 7 of the NPC patients. Samples were assessed for Tregs (CD4+FoxP3+, CD4+CD25+FoxP3+, CD8+FoxP3+, and CD8+CD25+FoxP3+) using multicolour flow cytometry, and XhoI mutation using polymerase chain reaction. Tregs proliferation was determined by Ki67 expression, and ex vivo proliferation was assessed after culturing PBMCs with IL-2 and anti-CD3/CD28 mAbs. The expression levels of five cytokines (IL-2, IL-10, IL-6, TNF-α, and IFN-ƴ) were evaluated using multiplex bead analysis. Tregs were generally found to be elevated in NPC patients. XhoI mutation was detected in both patients (39%) and controls (30%). The correlation between clinicopathological features and the frequency of circulating Tregs in the patients revealed no significant correlation for both CD4+ and CD8+ Tregs, even though Tregs levels were fairly higher in patients with stage IV compared with stage II/III cancer. In comparison with unstimulated cells, both CD4+ and CD8+ Tregs proportion heightened following the ex vivo culture of PBMCs. CD4+ and CD8+ Tregs in the PBMCs obtained from healthy controls responded better to ex vivo stimulation compared with those from patients. Of the cytokines, significant expression levels were observed for plasma IL-10 and TNF-α in patients compared with controls. The concentration of IL-10 was significantly lower in patients (10.47 ± 16.17 versus 16.12 ± 33.38, p = 0.0157). Meanwhile, plasma levels of TNF-α were higher in patients (p <0.0001) and were found to correlate with CD4+ Tregs in controls but not in patients. Correlation between IL-2 and CD8+CD25+FoxP3+ (r = -0.4670, p = 0.0438) Tregs was also observed. Data from this study demonstrates that targeting or developing therapies that specifically modulate Tregs number, trafficking, or activity, will be promising in NPC treatment. 2023-05 Thesis NonPeerReviewed application/pdf en http://eprints.usm.my/58895/1/AHMAD%20ADEBAYO%20IREKEOLA-24%20pages.pdf Irekeola, Ahmad Adebayo (2023) Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1 Xhoi mutation in nasopharyngeal carcinoma. PhD thesis, Universiti Sains Malaysia. |
| spellingShingle | QH573-671 Cytology R Medicine Irekeola, Ahmad Adebayo Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1 Xhoi mutation in nasopharyngeal carcinoma |
| title | Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1
Xhoi mutation in nasopharyngeal carcinoma |
| title_full | Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1
Xhoi mutation in nasopharyngeal carcinoma |
| title_fullStr | Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1
Xhoi mutation in nasopharyngeal carcinoma |
| title_full_unstemmed | Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1
Xhoi mutation in nasopharyngeal carcinoma |
| title_short | Elucidating the proportion of CD4+ and CD8+ regulatory t-cells, associated Cytokines, and epstein-barr virus’ LMP1
Xhoi mutation in nasopharyngeal carcinoma |
| title_sort | elucidating the proportion of cd4 and cd8 regulatory t cells associated cytokines and epstein barr virus lmp1 xhoi mutation in nasopharyngeal carcinoma |
| topic | QH573-671 Cytology R Medicine |
| url | http://eprints.usm.my/58895/1/AHMAD%20ADEBAYO%20IREKEOLA-24%20pages.pdf |
| work_keys_str_mv | AT irekeolaahmadadebayo elucidatingtheproportionofcd4andcd8regulatorytcellsassociatedcytokinesandepsteinbarrviruslmp1xhoimutationinnasopharyngealcarcinoma |