Fachry, Ade Wildan Rizky and Prof. Dr. dr. Kusworini H., M.Kes, Sp.PK (K) and Dr. dr. Hani Susianti, Sp.PK (K) (2023) Hubungan Antara Penanda Kostimulator sCD80 dan sCD86 terhadap Immune Risk Profile (IRP) Sel Limfosit T CD8+CD57+ pada Proses Penuaan Imun. Magister thesis, Universitas Brawijaya.
Abstract
Penuaan merupakan suatu fenomena kompleks yang ditandai dengan penurunan fisiologi tubuh secara progresif fungsi dan homeostasis jaringan yang dapat menyebabkan peningkatan kerentanan dan kematian. Seiring dengan bertambahnya usia, sistem imun tubuh akan mengalami proses penuaan yang ditandai dengan penurunan fungsi kekebalan tubuh secara progresif yang terkait dengan peningkatan frekuensi infeksi dan penyakit kronis. Penuaan sistem imun ditandai dengan suatu penanda yang disebut sebagai immune risk profile (IRP). Namun,IRP yang hanya dapat diamati dengan pemeriksaan flowsitometri yang cenderung tidak mudah, membutuhkan alat yang canggih dan ketrampilan serta tidak dapat dilakukan secara umum. Oleh karena itu diperlukan studi lebih lanjut untuk meneliti penanda prediktif penuaan sistem imun dengan metode yang lebih efektif dan efisien. Molekul kostimulator CD80 dan CD86 merupakan pasangan reseptor-ligan yang berperan penting dalam regulasi sistem imun melalui sinyal kostimulator untuk aktivasi, proliferasi dan promosis kelangsungan hidup sel T. Kedua penanda ini merupakan salah satu penanda penting dalam penuaan sistem imun dan penyakit autoimun. Molekul kostimulator ini memiliki bentuk terlarut dalam serum (sCD80 dan sCD86) dan telah diteliti memiliki peran dalam berbagai penyakit autoimun, terbentuk akibat pelepasan dari sel membran atau produksi alternatif saat proses splicing mRNA. Penelitian ini bertujuan untuk mengetahui adanya hubungan antara penanda IRP presentase sel limfosit T CD8+CD57+, baku emas deteksi immune aging dengan kadar sCD80 dan SCD86, sehingga diharapkan kedepannya kedua penanda terlarut ini dapat dikembangkan kit diagnostik immune aging. Penelitian ini merupakan studi kasus control, terdiri atas 87 responden penelitian, terbagi atas 40.2 % kelompok pasien LES, 36.8% lansia dan 23.0% individu muda. Presentase sel limfosit T CD8+CD57+ didapatkan dari PBMC dan diukur dengan teknik flowcytometri. PBMC yang telah dicuci dengan PBS, disentrifugasi, kemudian pellet ditambahkan dengan cell staining buffer dan dilakukan staining PerCP anti human CD8 (Biolegend Katalog no.344707) dan PE anti-human CD57 (Biolegend Katalog no. 359611). Pembacaan kuvet flowcytometri dilakukan dalam bentuk persentase sel, kemudian dihitung dengan BD Cell Quest software. Penanda sCD80 dan sCD86 pada serum diukur menggunakan metode ELISA. Kadar penanda kostimulator terlarut ini diukur dengan menggunakan Kit ELISA: Human Soluble CD80 (sCD80) ELISA Kit Mybiosource catalog number: MBS705535, dan HSCD 86 (sCD86) ELISA Kit MBS catalog number: MBS702416. Pembacaan Optical Density pada 450 nm, dengan pembaca Microelisa Stripplate. Data penelitian ini dianalisis menggunakan software IBM SPSS versi 23, menggunakan uji komparasi Kruska Wallis untuk menentukan perbedaan presentase sel limfosit T CD8+CD57+ serta kadar sCD80 dan Optical Density sCD86 pada ketiga ix kelompok penelitian. Uji Korelasi Spearman dilakukan untuk mengetahui hubungan antara penanda IRP presentase sel limfosit T CD8+CD57+ dengan kadar sCD80 dan sCD86. Pada penelitian ini didapatkan bahwa terdapat peningkatan sel limfosit T CD8+CD57+ pada lansia dan pasien LES, sedangkan terjadi penurunan pada individu muda. Kadar sCD80 dan Optical Density sCD86 diamati mengalami peningkatan pada kedua kelompok penuaan imun, yaitu lansia dan pasien LES. Uji korelasi Spearman menunjukkan bahwa penanda IP (immune risk profile) presentase sel limfosit T CD8+CD57+ memiliki hubungan yang tidak signifikan dengan penanda sCD80 (Kelompok Individu, p value= 0,819, r=-0,055; LES p value= 0,706, r=0,066 ; Lansia p value= 0,142, r=-0,266) dan sCD86 (Kelompok Individu, p value= 0,965, r=0,011; LES p value= 0,064, r=0,316 ; Lansia p value= 0,135, r=-0,270) pada penuaan system imun. Hal ini menunjukkan bahwa semakin tinggi presentase sel limfosit T CD8+CD57+ maka kadar sCD80 dan Optical density sCD86 akan semakin menurun. Begitu juga sebaliknya, semakin rendah presentase sel limfosit T CD8+CD57+maka kadar sCD80 dan Optical density sCD86 akan semakin meningkat. Hubungan antara IRP sel limfosit T CD8+CD57+dengan penanda sCD80 dan sCD86 pada penuaan sistem imun dapat dijelaskan melalui keterkaitannya dengan peningkatan stress oksidatif dan proses inflamaging, pada saatnya akan mengakibatkan perubahan populasi dan diversitas sel limfosit T, berupa: penurunan sel limfosit T CD8+CD57+dan peningkatan kadar penanda terlarut sCD80, serta peningkatan produksi SCD86 oleh APC. Sehingga dapat disimpulkan bahwa berdasarkan hail penelitian ini penanda terlarut sCD80 dan sCD86 dapat dijadikan salah satu biomarker prediktif penuaan sistem imun di masa depan. Namun mekanisme dan perananan penanda sCD80 dan sCD86 terkait hubungannya dengan sel limfosit T CD8+CD57+pada penuaan sistem imun masih perlu dikaji dan diteliti lebih lebih lanjut.
English Abstract
Aging is a complex phenomenon characterized by a progressive decline in body physiology, function and tissue homeostasis, which can lead to increased susceptibility and death. As we age, the body's immune system will experience an aging process characterized by a progressive decline in immune function associated with an increase in the frequency of infections and chronic diseases. Aging of the immune system is characterized by a marker called the immune risk profile (IRP). However, IRP that can only be observed by flowsitometry examination tends to be not easy, requires sophisticated tools and skills and cannot be done in general. Therefore, further studies are needed to examine predictive markers of aging of the immune system with more effective and efficient methods. CD80 and CD86 receptor-ligand molecules are receptor-ligand pairs that play an important role in immune system regulation through costimulator signaling for T cell activation, proliferation and promotion of survival. These costimulator molecules have dissolved forms in serum (sCD80 and sCD86) and have been studied to have roles in various autoimmune diseases, formed by detachment from cell membranes or alternative production during mRNA splicing. This study aims to determine the relationship between IRP markers of CD8+CD57+ T lymphocyte cell percentage, the gold standard for immune aging detection with sCD80 and SCD86 levels, so it is hoped that in the future these two dissolved markers can be developed immune aging diagnostic kits. This study is a case control study, consisting of 87 research respondents, divided into 40.2% of the LES patient group, 36.8% elderly and 23.0% young individuals. The percentage of CD8+CD57+ T lymphocyte cells was obtained from PBMC and measured by flowcytometry technique. PBMC that has been washed with PBS, centrifuged, then pellets are added with cell staining buffer and stained PerCP anti-human CD8 (Biolegend Catalog no.344707) and PE anti-human CD57 (Biolegend Catalog no. 359611). Cuvette flowcytometry readings are performed in the form of cell percentages, then calculated with BD Cell Quest software. Serum sCD80 and sCD86 markers were measured using the ELISA method. These dissolved costimulator marker levels were measured using ELISA Kit: Human Soluble CD80 (sCD80) ELISA Kit Mybiosource catalog number: MBS705535, and HSCD 86 (sCD86) ELISA Kit MBS catalog number: MBS702416. Optical Density reading at 450 nm, with Microelisa Stripplate reader. The data were analyzed using IBM SPSS software version 23, using the Kruska Wallis comparative test to determine differences in CD8+CD57+ T lymphocyte cell percentages and sCD80 and sCD86 Optical Density levels in the three study groups. The Spearman Correlation Test was performed to determine the relationship between IRP markers of CD8+CD57+ T lymphocyte cell percentage and sCD80 and sCD86 levels. In this study it was found that there was an increase in CD8+CD57+ T lymphocyte cells in the elderly and LES patients, while there was a decrease in young xi individuals. Levels of sCD80 and Optical Density of sCD86 were observed to increase in both groups of immune aging, namely elderly and LES patients. The Spearman correlation test showed that the IP (immune risk profile) marker of CD8+CD57+ T lymphocyte cell percentage had an insignificant association with the sCD80 marker (Individual Group, p value= 0.819, r=-0.055; LES p value= 0.706, r=0.066 ; Elderly p value = 0.142, r = -0.266) and sCD86 (Individual Group, p value = 0.965, r = 0.011; LES p value= 0.064, r=0.316 ; Elderly p value = 0.135, r = -0.270) in aging immune system. This shows that the higher the percentage of CD8+CD57+ T lymphocyte cells, the lower the sCD80 and sCD86 optical density levels. Vice versa, the lower the percentage of CD8+CD57+T lymphocyte cells, the higher the sCD80 and sCD86 optical density levels. The relationship between the IRP of CD8+CD57+ T lymphocyte cells and sCD80 and sCD86 markers in the aging immune system can be explained by its association with increased oxidative stress and inflamaging processes, which in turn will result in changes in the population and diversity of T lymphocytes, in the form of: decreased CD8+CD57+ T lymphocyte cells and increased levels of sCD80 soluble markers, as well as increased production of sCD86 by APC. So it can be concluded that based on this research, soluble markers sCD80 and sCD86 can be used as one of the predictive biomarkers of aging of the immune system in the future. However, the mechanism and role of sCD80 and sCD86 markers related to their relationship with CD8+CD57+ T lymphocyte cells in aging immune systems still need to be studied and investigated further.
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | 0423060114 |
| Subjects: | 600 Technology (Applied sciences) > 616 Diseases > 616.02 Special topics of disease > 616.025 Medical emergencies / Emergency medicine / Emergency nursing / Triage (Medicine) |
| Divisions: | S2/S3 > Magister Ilmu Biomedis, Fakultas Kedokteran |
| Depositing User: | Endang Susworini |
| Date Deposited: | 14 Dec 2023 04:16 |
| Last Modified: | 14 Dec 2023 04:16 |
| URI: | http://repository.ub.ac.id/id/eprint/205425 |
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