Identification of peptides for immune activation and responses in Thai patients with systemic sclerosis
1
Issued Date
2020
Copyright Date
2020
Resource Type
Language
eng
File Type
application/pdf
No. of Pages/File Size
xv, 98 leaves : ill.
Access Rights
open access
Rights
ผลงานนี้เป็นลิขสิทธิ์ของมหาวิทยาลัยมหิดล ขอสงวนไว้สำหรับเพื่อการศึกษาเท่านั้น ต้องอ้างอิงแหล่งที่มา ห้ามดัดแปลงเนื้อหา และห้ามนำไปใช้เพื่อการค้า
Rights Holder(s)
Mahidol University
Bibliographic Citation
Thesis (M.Sc. (Tropical Medicine))--Mahidol University, 2020
Suggested Citation
Oranit Likhit Identification of peptides for immune activation and responses in Thai patients with systemic sclerosis. Thesis (M.Sc. (Tropical Medicine))--Mahidol University, 2020. Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/114322
Title
Identification of peptides for immune activation and responses in Thai patients with systemic sclerosis
Author(s)
Abstract
Systemic sclerosis (SSc), or scleroderma, is a chronic autoimmune disease involving a multisystem disorder characterized by skin and visceral organ fibrosis (such as heart, lungs, kidneys, gastrointestinal tract, and musculoskeletal systems), as well as an immune activation by the production of disease-specific autoantibodies in patients’ sera. There are two types of SSc
limited systemic sclerosis (lcSSc) and diffused systemic sclerosis (dcSSc). LcSSc is a mild form with minimal skin involvement while dcSSc is a severe type with rapid widespread skin and/or internal involvement, resulting in severe disability and death. In general, mortality and morbidity are directly related to patients with fibrotic and microvascular alterations. The etiology of SSc remains unknown but genetics, environmental factors, and immunological abnormalities have been found to contribute to the disease development. This study aims to investigate the role of CD4 + and CD8 + T cells on pathogenesis of systemic sclerosis. The peripheral blood mononuclear cells (PBMCs) isolated from SSc patients and healthy controls were stimulated with peptides derived from DNA topoisomerase proteins, and T cell responses were determined by intracellular cytokine staining (ICS) for interferon gamma (IFN-γ) and interleukin-2 (IL-2) production by CD4 + and CD8 + T cells. For the SSc patients, the high levels of IFN-γ (range 3.4–23.49, mean = 11.18) and IL-2 (range 4.88–24.58, mean = 12.12) secreting CD8 + T cells were detected while the lower levels of these cytokines were produced from CD4 + T cells (range 1.28–21.03, mean = 5.43
IFN-γ, range 2.45–24.58, mean = 6.81
IL-2. We did not observe the significant levels of cytokine-secreting CD4 + and CD8 + T cells in the healthy control groups. These results suggest that the cytokine production by T cells following stimulation with peptides derived from DNA topoisomerase protein is specific for the development of systemic sclerosis. For the exploration to identify the possible epitope to specifically related with SSc diseases, from the stimulated individual peptides experiment results showed that CSLRVEHINLHPELD (sPep3
15 amino acids
position 505–519 of DNA Topoisomerase I protein) was the optimal epitope, which induced T cells isolated from the SSc patients to secrete the highest level of cytokine production. We further investigated the association of the T cell responses and HLA haplotypes in the SSc patients. In this study, the most common HLA haplotypes detected in the SSc patients were DRB1*12:02, DRB1*15:01, DRB1*15:02, and DRB5*01:08. However, we did not observe a significant correlation between these common HLA haplotypes and T cell responses in the SSc patients. The knowledge gained from this study should pave the way for a better understanding of immunopathology in the SSc patients and to apply this knowledge for the development of the diagnostic tools and specific treatments for the SSc in the future.
limited systemic sclerosis (lcSSc) and diffused systemic sclerosis (dcSSc). LcSSc is a mild form with minimal skin involvement while dcSSc is a severe type with rapid widespread skin and/or internal involvement, resulting in severe disability and death. In general, mortality and morbidity are directly related to patients with fibrotic and microvascular alterations. The etiology of SSc remains unknown but genetics, environmental factors, and immunological abnormalities have been found to contribute to the disease development. This study aims to investigate the role of CD4 + and CD8 + T cells on pathogenesis of systemic sclerosis. The peripheral blood mononuclear cells (PBMCs) isolated from SSc patients and healthy controls were stimulated with peptides derived from DNA topoisomerase proteins, and T cell responses were determined by intracellular cytokine staining (ICS) for interferon gamma (IFN-γ) and interleukin-2 (IL-2) production by CD4 + and CD8 + T cells. For the SSc patients, the high levels of IFN-γ (range 3.4–23.49, mean = 11.18) and IL-2 (range 4.88–24.58, mean = 12.12) secreting CD8 + T cells were detected while the lower levels of these cytokines were produced from CD4 + T cells (range 1.28–21.03, mean = 5.43
IFN-γ, range 2.45–24.58, mean = 6.81
IL-2. We did not observe the significant levels of cytokine-secreting CD4 + and CD8 + T cells in the healthy control groups. These results suggest that the cytokine production by T cells following stimulation with peptides derived from DNA topoisomerase protein is specific for the development of systemic sclerosis. For the exploration to identify the possible epitope to specifically related with SSc diseases, from the stimulated individual peptides experiment results showed that CSLRVEHINLHPELD (sPep3
15 amino acids
position 505–519 of DNA Topoisomerase I protein) was the optimal epitope, which induced T cells isolated from the SSc patients to secrete the highest level of cytokine production. We further investigated the association of the T cell responses and HLA haplotypes in the SSc patients. In this study, the most common HLA haplotypes detected in the SSc patients were DRB1*12:02, DRB1*15:01, DRB1*15:02, and DRB5*01:08. However, we did not observe a significant correlation between these common HLA haplotypes and T cell responses in the SSc patients. The knowledge gained from this study should pave the way for a better understanding of immunopathology in the SSc patients and to apply this knowledge for the development of the diagnostic tools and specific treatments for the SSc in the future.
Degree Name
Master of Science
Degree Level
Master's degree
Degree Department
Faculty of Tropical Medicine
Degree Discipline
Tropical Medicine
Degree Grantor(s)
Mahidol University