Turk J Immunol. 2017; 5(1): 13-19 | DOI: 10.25002/tji.2017.544

Immunomodulatory Effect of Propolis Extract on Granzyme Expression in CD8+ and CD4+CD25+ T Cells

Zauhani Kusnul¹, Pudji Rahayu², Muhaimin Rifai³, Edi Widjajanto^4
1Bahrul Ulum Nursing Academy, Jombang, Indonesia, jombang, Indonesia
²Saiful Anwar General Hospital, Faculty of Medicine, University of Brawijaya, Malang, Indonesia, Department of Ear, Nose, and Neck, Malang, Indonesia
³Faculty of Sciences, University of Brawijaya, Malang, Indonesia, Department of Biology, Malang, Indonesia
⁴Saiful Anwar General Hospital, Faculty of Medicine, University of Brawijaya, Malang, Indonesia, Department of Clinical Pathology, Malang, Indonesia

Objective: The purpose of this study was to explore the effect of propolis extract on CD8+ and CD4+CD25+ regulatory T cell populations and granzyme expression in both cell populations, as propolis has been suggested as an immunomodulatory agent. Materials and Methods: PBMC containing 1x106 cells/ml from cervical cancer stage 3 patients were isolated and cultured with and without propolis extract. The cells were plated in a 24-well plate with RPMI medium supplemented with 10% FBS, 10 U/ml penicillin, 100 U/ml streptomycin, 2 mM L-glutamine and 50 µM 2-ME. Propolis extract was added (5, 10, 25 and 50 µg/ml) to PBMC culture. Negative control was made from PBMC culture without propolis extract for comparison. The cells were incubated for 48 hours in a 5% CO2 incubator at 37ºC. The proportion of CD8+ and CD4+CD25+ regulatory T cells that expressed granzyme were measured by flow cytometry. Results: The results obtained that the level of CD4+, CD4+CD25+, and CD8+ T cells were not markedly depleted in propolis treatment in all doses compared to control group. The percentage of CD8+Granzyme+ and CD4+CD25+Granzyme+ were significantly different in groups of propolis treatment compared to control group. Conclusion: In summary, propolis extract may serve as an immunomodulatory agent in cervical cancer patients. Propolis extract could modulate the expression CD4+CD25+ T cells expressing granzyme and CD8+ T cells expressing granzyme while it has no effect on CD4+, CD4+CD25+, and CD8+ T cells.

Keywords: CD8+ T cells, CD4+CD25+ regulatory T cells, granzyme, propolis

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Propolis Ekstresinin CD8+ ve CD4+CD25+ T Hücrelerindeki Granzim Ýfadesi Üzerine Olan Baðýþýklýk Düzenleyici Etkileri

Zauhani Kusnul¹, Pudji Rahayu², Muhaimin Rifai³, Edi Widjajanto^4
1Bahrul Ulum Nursing Academy, Jombang, Indonesia, jombang, Indonesia
²Saiful Anwar General Hospital, Faculty of Medicine, University of Brawijaya, Malang, Indonesia, Department of Ear, Nose, and Neck, Malang, Indonesia
³Faculty of Sciences, University of Brawijaya, Malang, Indonesia, Department of Biology, Malang, Indonesia
⁴Saiful Anwar General Hospital, Faculty of Medicine, University of Brawijaya, Malang, Indonesia, Department of Clinical Pathology, Malang, Indonesia

Amaç: Çalýþmanýn amacý, propolis ekstresinin baðýþýklýk düzenleyici bir madde olarak CD8+ ve CD4+CD25+ düzenleyici T hücrelerindeki granzim ifadesi üzerine olan etkisini incelemektir. Gereçler ve Yöntemler: Evre 3 serviks kanserli hastalardan elde edilen 10(6) hücre/mL konsantrasyonundaki Periferik Mononükleer Kan Hücreleri(PMKH) izole edilerek propolis eklenen ve eklenmeyen kaplarda kültüre edildi. Hücreler, %10 FBS, 10 U/ml penisilin, 100 U/mL streptomisin, 2mM L-glutamin ve 50 mikroM 2-ME içeren 24 kuyulu kültür kaplarýna paylaþtýrýldý. Bir PMKH popülasyonu Propolis uygulanmadan kontrol grubu olarak oluþturuldu. Hücreler %5 CO2 içeren kuluçka makinesinde(inkübatör) 48 saat kültüre edildi. Granzim ifade eden CD8+, CD4+CD25+ düzenleyici T lenfositleri akan hücre ölçer ile irdelendi. Bulgular: Propolis uygulanan CD4+, CD4+CD25+ ve CD8+ T lenfositlerinin sayýlarý anlamlý ölçüde azalmadý. CD8+Granzim+ ve CD4+CD25+Granzim+ hücrelerin sayýlarý, propolis uygulanan hastalarda farklý idi. Sonuç: Sonuç olarak, propolis ekstresi serviks kanserli hastalarda baðýþýklýðý deðiþtiren bir madde olarak etki etmektedir. Ekstre, CD4+CD25+ T lenfositleri ile CD8+ T lenfositlerinde granzim ifadesini deðiþtirmekte, ancak ekstrenin CD4+, CD+CD25+ ve CD8+ T lenfositlerinde granzim ifadesine etkisi görülmemektedir.

Anahtar Kelimeler: CD8+ T Hücreleri, CD4+CD25+ düzenleyici T Hücreleri, granzim, propolis

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