Profil Peripheral Blood Mononuclear Cells (PBMC) Pasien dengan Berbagai Usia Menggunakan Flow Cytometry di Klinik Hayandra

Authors

  • Masayu Nadhira Universitas Al Azhar Indonesia
  • Riris Lindiawati Puspitasari Universitas Al Azhar Indonesia
  • Karina F Moegni Yayasan Hayandra Peduli
  • Imam Rosadi Yayasan Hayandra Peduli
  • Iis Rosliana Yayasan Hayandra Peduli

DOI:

https://doi.org/10.36722/sst.v4i4.312

Abstract

Abstrak - Profil Peripheral Blood Mononuclear Cells (PBMC) setiap orang dapat berbeda-beda. Salah satu faktor yang diduga mempengaruhi adanya perbedaan tersebut adalah usia. Manusia berusia lanjut lebih mudah terserang penyakit karena terjadinya penurunan fungsi dan jumlah sel imun. Oleh karena itu, perlu dilakukan analisis komposisi PBMC untuk mengetahui profil PBMC seseorang berdasarkan usia, sehingga dapat diketahui jumlah sel imun dalam tubuh manusia. Tujuan dari penelitian ini adalah untuk menganalisis profil PBMC pasien dengan berbagai usia di Klinik Hayandra menggunakan flow cytometry. Manfaat dilakukannya kegiatan penelitian ini adalah diperolehnya informasi mengenai profil PBMC pasien di Klinik Hayandra dan kaitannya dengan usia. Metode yang dilakukan diantaranya adalah isolasi PBMC, proses pewarnaan PBMC, dan penentuan populasi sel menggunakan flow cytometry. Cluster of Differentiation (CD) yang digunakan pada kegiatan penelitian ini diantaranya adalah CD3, CD19, CD8, CD56, dan CD314 atau disebut juga NKG2D (Natural Killer Group 2 member D). Hasil yang diperoleh pada kegiatan penelitian ini adalah ditemukannya korelasi negatif yang bermakna antara persentase jumlah sel limfosit T sitotoksik (CD3+/CD8+) dengan usia dan ditemukannya korelasi positif yang bermakna antara persentase jumlah sel NKT (CD3+/CD56+) dengan usia. Dengan demikian usia pasien dapat mempengaruhi jumlah sel imun pasien.

Abstract - Peripheral Blood Mononuclear Cells (PBMC) profiles of each person can different. One of the factors that allegedly affects the difference is age. Elderly human are more susceptible attacked disease due to decreased function and number of immune cells. Therefore, it is necessary to do analyze PBMC composition to know the PBMC profile of person, so that can be known the number of immune cells in the human body. The purpose of this research is to analyze the PBMC profile of patients with various ages in the Hayandra Clinic using flow cytometry. The benefit of doing this practical work is obtained information about the PBMC profile of patients in the Hayandra Clinic and its relation to age. Methods which are conducted among others are PBMC isolation, PBMC staining process, running flow cytometry, and determination of cell population. Cluster of Differentiation (CD) used in this practical work among others are CD3, CD19, CD8, CD56, and CD314 or also called NKG2D (Natural Killer Group 2 member D). The results obtained in this research were the found of significant negative correlation between percentage of cytotoxic T cell lymphocytes numbers (CD3+/CD8+) with age and found a significant positive correlation between percentage of NKT cell numbers (CD3+/CD56+) with age. So it can be concluded that the patient's age can affect the number of the patient immune cells.

Keywords - Peripheral Blood Mononuclear Cells (PBMC), flow cytometry, age, Cluster of Differentiation (CD)

Author Biographies

Masayu Nadhira, Universitas Al Azhar Indonesia

Program Studi Biologi, Fakultas Sains dan Teknologi

Riris Lindiawati Puspitasari, Universitas Al Azhar Indonesia

Program Studi Biologi, Fakultas Sains dan Teknologi

Karina F Moegni, Yayasan Hayandra Peduli

Klinik Hayandra

Imam Rosadi, Yayasan Hayandra Peduli

Klinik Hayandra

Iis Rosliana, Yayasan Hayandra Peduli

Klinik Hayandra

References

Y. Arina, "Pengaruh Aging Terhadap Sistem Imun," JKM, vol. 3, no.

, pp. 53-57, 2003.

Zenbio, "Peripheral Blood Mononuclear Cells (PBMCs)," ZenBio,

Inc, 2016. [Online]. Available: http://www.zen-

bio.com/products/cells/pbmcs.php. [Diakses pada 1 Februari

.

S. E, Anatomi dan Fisiologi untuk Pemula, Jakarta: Penerbit Buku

Kedokteran EGC, 2004.

R. Lane, "Four Downstream Applications for Isolated Peripheral

Blood Mononuclear Cells," Folio Conversant, 2014 November 2014.

[Online]. Available:https://www.dls.com/biopharma/blog/4-

downstream-applications-of-pbmc-isolation-from-whole-blood.

[Diakses pada 15 Maret 2018].

A. Unitly and D. Sahertian, "Deteksi Kandungan Antioksidan

Superoksida Dismutase (SOD) pada Organ Ginjal Tikus Ratus

Norvegicus Dengan Pewarnaan Imunohistokimia," in Seminar

Nasional Basic Science II, Malang, 2010.

T. Sun, Flow Cytometry, Immunohistochemistry, and Molecular

Genetics for Hematologic Neoplasms Second Editions, USA:

Lippincott Williams dan Wilkins, 2012.

I. U. S, T. Teguh and B. Mulyono, "Aplikasi Flow Cytometry di

Laboratorium," Berkala Kesehatan Klinik, no. 2, pp. 115-125,

F. Nathalia, "Perbandingan Nilai Hematologi Antara Medical Check

Up (MCU) di Rumah Sakit Puri Medika Jakarta dengan Nilai

Rujukan alat Sysmex XS-800i.[thesis]," Universitas Kristen

Maranatha, Bandung, 2014.

N. Brown and C. Wittwer, "Flow cytometry: principles and clinical

applications in hematology.," Clin Chem, vol. 46, no. 2, pp. 1221-

, 2000.

R. Robinson and S. Pellenz, "What is flow cytometry (FACS

analysis)?," antibodies online.com, 12 juni 2013. [Online].

Available: https://www.antibodies-

online.com/resources/17/1247/what-is-flow-cytometry-facs-

analysis/. [Diakses 2018 April 01].

J. Segen, Concise dictinory of, New York: The Mc Graw-Hill

Companies, Inc, 2002.

W. Sandhika, R. M. Novarina and T. Setyaningrum, "Pemeriksaan

Imunohistokimia untuk Mengungkap Patogenesis Vitiligo," CDK-

, vol. 43, no. 10, pp. 742-746, 2016.

K. Wang, G. Wei and a. D. Liu, "CD19: a biomarker for B cell

development, lymphoma diagnosis and therapy.," Hematol Oncol,

vol. 1, no. 36, 2012.

F. Naeim, P. Rao, S. Song and R. Phan, Atlas of Hematopathology,

Tokyo: Academic Press, 2013.

D. E. Meyers, S. Thakur, C. M. Thirukkumaran and D. G. Morris,

"Oncolytic virotherapy as an immunotherapeutic strategyfor

multiple myeloma," Blood Cancer Journal, vol. 7, no. 12, 2017.

Y. Jing, S. Gravenstein, N. Chaganty, N. Chen, K. Lyerly, S. Joyce

and Y. Deng, "Aging is associated with a rapid decline in

frequency, alterations in subset composition, and enhanced Th2

response in CD1d-restricted NKT cells from human peripheral

blood.," Exp Gerontol, vol. 42, no. 8, pp. 719-732, 2007.

C. Corkum, D. Ings, C. Burgess, S. Karwowska, W. Kroll and T.

Michalak, "Immune cell subsets and their gene expression

profiles from human PBMC isolated by Vacutainer Cell Preparation

Tube (CPTâ„¢) and standard density gradient.," BMC Immonual,

vol. 16, no. 48, 2015.

L. Doiron, "5 Types of Mononuclear Cells and How They Differ,"

Folio Conversant, 10 May 2016. [Online]. Available:

https://www.dls.com/biopharma/blog/5-types-of-mononuclear-

cells-and-how-they-differ. [Diakses pada 24 Februari 2018].

C. R. Kleiveland, "Peripheral Blood Mononuclear Cells," in The

Impact of Food Bioactives on Health: in vitro and ex vivo models,

US, National Library of Medcine, 2015, pp. 161-167.

L. Doiran, "3 Reasons Whole Blood is Necessary for PBMC

Isolation," Folio Conversant, 31 Maret 2015. [Online].

Available:http:www.conversantbio.com/blog/3-reasonswhole-

blood-is-necessary-forpbmcisolation/. [Diakses pada 06 Mei

.

H. Putri, "Preparasi Sampel Untuk Siklus Sel Dengan Metode

Flowcytometry," Fakultas Farmasi UGM, Yogyakarta, 2014.

E. O'Donnell, D. Ernst and R. Hingorani, "Multiparameter Flow

Cytometry: Advances in High Resolution Analysis," Imune

Network, vol. 13, pp. 43-54, 2013.

Bio-Rad Laboratories, "Data Analysis in Flow Cytometry," Bio-

Rad Laboratories, 2016. [Online]. Available: https;//www.bio-

radantibodies.com/flow-cytometry-gating-strategies.html/.

[Diakses pada 11 April 2018].

C. Paul, "An Introduction to Gating in Flow Cytometry," Bitesize

Bio, 2015. [Online].Available:https://bitesizebio.com/21596/an-

introduction-to-gating-in-flow-cytometry/. [Diakses pada 11 April

.

Sugiyono, Metode Penelitian Pendidikan Pendekatan Kuantitatif,

Kualitatif , dan R&D, Bandung: Alfabeta, 2014.

Fatmah, "Respons Imunitas yang Rendah pada Tubuh Manusia

Usia Lanjut," Makara Kesehatan, vol. 10, no. 1, pp. 48-50, 2006.

Andrea, "Natural Killer Cell subtypes and markers in human

PBMC," PBMC Basics, 14 Maret 2013. [Online].

Available:https://technical.sanguinebio.com/natural-killer-cell-

subtypes-and-markers-in-human-pbmc/. [Diakses pada 24

February 2018].

M. L. Garff-Tavernier, V. Beziat, DecocaJ, V. Siguret, F.

Gandibakhch, E. Pautas, P. Debre, H. Merle-Beral and Vieillard,

"Human NK cells display major phenotypic and functional changes

over the life span," Aging Cell, vol. 9, no. 4, pp. 527-535, 2010.

E. P. Remarque, "T cell immunosenescence and its clinical

relevance in man," Rev. Clin. Gerontology, vol. 8, pp. 5-14, 1998.

Y. Jiang, X. Cui, C. Cui, J. Zhang, F. Zhou, Z. Zhang, Y. Fu, J. Xu,

Z. Chu, J. Liu, X. Han, C. Liao, Y. Wang, Y. Cao and H. Shang,

"The Function of CD3+CD56+ NKT-Like Cells in HIV-Infected

Individuals," Biomed Res Int, no. 863625, 2014.

B. J. Terabe M, "The role of NKT cells in tumor immunity.," Adv

Cancer Res, no. 101, pp. 277-348, 2008.

Y. Chen and H. Liao, "NK/NKT Cells And Aging," Internasional

Journal of Gerontology, vol. 1, no. 2, pp. 65-69, 2007.

P. Sansoni, A. Cossariza, V. Brianti, F. Fagnoni, G. Sneli, D. Monti,

A. Marcato, G. Passeri, C. Ortalani, E. Forti, U. Fagiolo, M. Passed

and C. Franceschi, "Lymphocyte subsets and natural killer cell

activity in healthy old people and centenarians," Blood 82, no.

-2773, 1993.

S. M. Nerlans, I. Ria, H. Alexander and T. Moris, "Changes in

natural killer cells, the CD57CD8 subset, and realted cytokines in

healthy aging," J. Clin Immunol, no. 18, pp. 31-38, 1998.

M. Gharagozolo, A. Rezai, H. Kalantari, A. Bahador, N.

Hassannejad, M. Maracy, N. Noury, M. Sedghi, H. Ghazanfari and

B. Bayat, "Decline in peripheral blood NKG2D+CD 3+CD 56+ NKT

cells in colorectal cancer patient," Bratislava Medical Journal, vol.

, no. 1, pp. 6-11, 2018.

R. L. Puspitasari, A. Boediono and F. Sandra, "Conditioned

medium dari kultur primer sel syaraf Mus musculus," in Seminar

Nasional X Pendidikan Biologi FKIP UNS, Solo, 2013.

S. JC, Concise dictionary of modern medcine, New York: The

McGraw-Hill Companies, 2002.

Mendez-David, Z. El-Ali, R. Hen, B. Fallissard and E. Corruble, "A

method for biomarker measurements in peripheral blood

mononuclear cells isolated from anxious and depressed mice: β-

arrestin 1 protein levels in depression and treatment.," Front

Pharmacol, vol. 4, no. 124, 2013.

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Published

2018-12-13

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JURNAL AL-AZHAR INDONESIA SERI SAINS DAN TEKNOLOGI