Journals / Clinical and Experimental Health Sciences / 2011 / Cilt: 1 - Sayı: 3

Lymphocyte proliferation in common variable immunodeficiency (CVID) patients by carboxyfluorescein succinimidylester (CFSE)

Sık değişken immün yetmezlik hastalarında Carboxyfluorescein succinimidylester (CFSE) yöntemi ile lenfosit proliferasyonu

Pages
171–177
Publication Date
2014-01-31
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Conclusion: In CVID patients T cell and B cells proliferation seem to be normal.

📌 3 Key Takeaways

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Objective & Hypothesis:Objective: Common variable immunodeficiency (CVID) is a heterogeneous disease in the group of predominantly antibody deficiencies, which is defined by hypogammaglobulinemia and normal or low level of B cells, and characterized by increased susceptibility to recurrent bacterial infections, autoimmune disorders, and malignancies.
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Key Findings & Data:Method: The proliferation of CD19 + , CD21 + , CD27 + B cells, and CD3 + , CD4 + , CD8 + T cells were investigated by Carboxyfluorescein Succinimidyl Ester (CFSE) method.
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Conclusion & Impact:Conclusion: In CVID patients T cell and B cells proliferation seem to be normal.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:SDİY · CFSE · Lenfosit proliferasyonu

Abstract

Objective: Common variable immunodeficiency (CVID) is a heterogeneous disease in the group of predominantly antibody deficiencies, which is defined by hypogammaglobulinemia and normal or low level of B cells, and characterized by increased susceptibility to recurrent bacterial infections, autoimmune disorders, and malignancies. In this study, we aimed to investigate the proliferation of the B and T lymphocytes in patients with CVID. Method: The proliferation of CD19 + , CD21 + , CD27 + B cells, and CD3 + , CD4 + , CD8 + T cells were investigated by Carboxyfluorescein Succinimidyl Ester (CFSE) method. CVID patients (n=13) and control group (n=5) were enrolled in the study. B and T cells were isolated from periferal blood samples with the negative selection procedure called Rosette Sep andthe isolated lymphocytes were stained in dark using CFSE. The cells were stimulated with anti- CD2, CD3, CD28 (CDmix) and cultured for four days in order to determine T cell proliferation. The proliferation of CD3 + , CD4 + , CD8 + T cells was analyzed by flow cytometry. Isolated B cells were cultured for four days along with anti-IgM+IL-4. The proliferation of CD19 + , CD27 + and CD21 + B cells were analyzed with flow cytometry. Results: First proliferation of T lymphocytes and CD19 + B lymphocytes and second proliferation of CD4 + T lymphocytes after stimulation were increased in CVID patients compared to healthy controls. Conclusion: In CVID patients T cell and B cells proliferation seem to be normal.

Özet

Amaç: Sık Değişken İmmün Yetmezlik (SDİY); hipogamaglobülinemi ve normal / düşük B hücre ile tanımlanan antikor eksikliği hastalıkları grubundandır ve tekrarlayan bakteriyel infeksiyonlar, otoimmün hastalıklar ve malignite ile karakterize edilmektedir. Bu çalışmada, SDİY hastalarındaki T ve B lenfositlerinin proliferasyonlarının incelenmesi amaçlanmıştır. Yöntem: T ve B hücre alt gruplarında hastalığın immünopatogenezinde önemli rol oynadığı düşünülen CD19 + , CD21 + , CD27 + B hücreleri ve CD3 + , CD4 + , CD8 + T hücrelerinin Carboxyfluorescein SuccinimidylEster (CFSE) ile hücre proliferasyon yanıtlarına bakıldı. Çalışmaya SDİY tanısı almış hastalar (n=13) ve kontrol grubu için SDİY hastalarıyla yaş uyumu olan sağlıklı bireyler (n=5) alındı. Tüm deneklerden alınan kan örneklerine, negatif seçim yöntemi (Rosette Sep) ile T ve B hücre izolasyonu yapıldı. Daha sonra izole edilen lenfositler CFSE-FITC ile karanlık ortamda işaretlendi ve T hücre proliferasyonu için hücreler anti-CD2, anti-CD3, anti-CD28 (CDmix) ile uyarılarak 4 günlük kültürleri yapıldı. Kültür sonunda hücreler anti-CD3-PE, anti-CD4-PE, anti-CD8-APC ile boyandıktan sonra CD3 + , CD4 + , CD8 + T hücrelerinin proliferasyonu akım sitometrisi ile bakıldı. B hücre proliferasyonu için hücreler anti-IgM + IL-4 ile 4 gün kültür edildikten sonra anti-CD19-APC-Cy7, anti- CD27-PE, anti-CD21-APC ile boyandı ve CD19 + , CD27 + ve CD21 + B hücrelerinin proliferasyonu akım sitometrisi ile bakıldı. Bulgular: T lenfositlerinin ve CD19 + B hücrelerinin proliferasyonlarının 1. bölünmesinde ve CD4 + T hücrelerinin 2. bölünmesinde, CDmix uyaranlı SDİY’li hastalarda, CDmix uyaranlı kontrolden anlamlı olarak daha fazla prolifere olduğu gözlendi. Sonuç: T hücreleri ve B hücrelerinin proliferasyonlarının SDİY hastalarında normal olduğu gösterildi.

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Cite This Article

İzgi, A., Akkoç, T., Öğülür, İ., Tevetoğlu, A., Aydıner, E. K., Barış, S., Bahçeciler, N., Barlan, I. (2014). Lymphocyte proliferation in common variable immunodeficiency (CVID) patients by carboxyfluorescein succinimidylester (CFSE). Clinical and Experimental Health Sciences, Cilt: 1 - Sayı: 3, 171–177. https://ajindex.com/articles/481910

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