Doğal öldürücü (NK) hücre düşüklüğü olan hastalarda, NK hücre sitotoksitelerinin ve FcγRIIIA gen mutasyonlarının araştırılması
Yükleniyor...
Dosyalar
Tarih
2020
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Necmettin Erbakan Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Doğal öldürücü hücreler (natural killer, NK), organizmayı enfeksiyonlara ve kansere karşı savunmada önemli rol oynayan hücrelerdir. NK hücrelerini organizmada iki önemli görevi bulunmaktadır. Bunlardan birincisi, enfeksiyona neden olan mikroorganizmalara karşı direkt sitotoksik etki, ikincisi ise FcγRIIIA geni aracılığıyla gerçekleşen antikor bağımlı hücresel sitotoksisite (ADCC)'dir. NK hücrelerinin sayı veya fonksiyonunda bozukluk(lar), NK hücre eksikliği/bozukluğu (NKD) olarak adlandırılmaktadır. Bu çalışmada, klinik olarak NKD düşünülen hastalar NK hücre alt grupları, NK hücre sitotoksisitesi ve FcγRIIIA gen mutasyon(lar)u açısından araştırılması ve klinik, laboratuvar ve fonksiyonel analiz sonuçları ile FcγRIIIA gen mutasyon(lar)u arasındaki ilişkinin değerlendirilmesi amaçlandı. Çalışmaya Necmettin Erbakan Üniversitesi Meram Tıp Fakültesi Çocuk Allerji ve İmmünoloji polikliniğine ilk defa başvuran hastalar ile daha önceden takipli 10 hasta ve 7 sağlıklı bireyden oluşan kontrol grubu dahil edildi. Tüm hasta ve kontrol kan örneklerinden, periferik kan lenfosit oranları ve NK hücre alt grupları oranları akım sitometri yöntemiyle, NK hücrelerinin fonksiyonları ise sitotoksisite testi ile değerlendirildikten sonra yeni nesil dizileme yöntemi ile FcγRIIIA gen mutasyonları araştırıldı. Periferik kan akım sitometri yöntemi ile değerlendirilen NK hücre alt grup analizinde CD56brightCD16neg, CD56brightCD16int ve CD56dimCD16hi olmak üzere 3 farklı NK hücre alt grubu belirlendi. Hastaların NK hücre alt grupları kontrol grubu ile karşılaştırıldığında CD56brightCD16neg hücre oranında statistiksel olarak anlamlı bir farklılık tespit edilmedi. CD56brightCD16int ve CD56dimCD16hi hücre oranları kontrol grubu ile karşılaştırıldığında hastalarda anlamlı derecede düşük olduğu tespit edildi. NK hücre sitotoksisitesi, K562 lizisine bağlı yöntem ile değerlendirildi. NK hücre sitotoksisite analizi sonucunda hasta ile kontroller arasında K562 miktarındaki oransal olarak azalmanın istatistiksel olarak anlamlı olduğu tespit edildi (p<0,001). Hastalardaki K562 hücre azalma oranı kontrollere kıyasla istatistiksel olarak daha az azaldığı tespit edildi. Yeni nesil dizileme yöntemi ile FcγRIIIA geninin tüm exom analizinde, ekzon 4 (rs396991) ve ekzon 3 (rs10127939)'de polimorfizmler olduğu belirlendi. Ayrıca dizileme sonucunda FcγRIIIA ile FcγRIIIB ile benzerlik gösteren olası kimerik bir bölge (ekzon 1 ile ekzon 2 ile ekzon 1 arasındaki intron bölgesi) tespit edildi. Sonuç olarak bu tez çalışmasında FNKD düşünülen hastalar ileri fonksiyonel ve genetik analizler ile değerlendirildi. Hastalarda NK sayı ve fonksiyonları yönünden farklılıklar tespit edildi. FcγRIIIA geni ile ilgili tespit edilen polimorfizmler Türkiye popülasyonunda yapılmış ilk değerlendirme niteliğinde olup, NK hücre araştırmaları alanında yapılacak yeni çalışmalara ışık tutması beklenmektedir.
Natural killer cells (NK) are cells that play an important role in defending the organism against infections and cancer. NK cells have two important functions in the organism. The first is the direct cytotoxic effect against microorganisms that cause infection, Secondly, it is an antibodydependent cellular cytotoxicity (ADCC) that is mediated by the FcγRIIIA gene product. Considering the important role of NK cells, disorder(s) in the NK cell number or function cause disease. This disease originating from NK cells is called NK cell deficiency / disorder (NKD). From this point, NK cell subgroups, NK cell cytotoxicity and FcγRIIIA gene mutation(s) were investigated. The aim of this study was to evaluate the relationship between clinical, laboratory and functional analysis and FcγRIIIA gene mutation(s). The study group consisted of 10 patients who were admitted to Necmettin Erbakan University Meram Medical Faculty Pediatric Allergy and Immunology Policlinic for the first time and previously monitored and consisting of 7 healthy individuals were included in the control group. All patients and controls were evaluated for the determination of peripheral blood lymphocyte ratios and NK cell subgroups and cytotoxicity tests were performed to evaluate the functions of NK cells. FcγRIIIA gene mutations were investigated by next generation sequencing method. Three different NK cell subgroups, CD56brightCD16neg, CD56brightCD16int and CD56dimCD16hi were identified in the NK cell subgroup analysis evaluated by peripheral blood flow cytometry. No statistically significant difference was found in the ratio of CD56brightCD16neg cells when NK cell subsets of the patients were compared with the control group. CD56brightCD16int and CD56dimCD16hi cell ratios were found to be significantly lower in patients compared to the control group. NK cell cytotoxicity was evaluated by K562 lysis dependent method. As a result of NK cell cytotoxicity analysis, proportional decrease of K562 amount between patient and controls was found to be statistically significant (p<0,001). It was found that the rate of K562 cell reduction in the patients decreased statistically less than the controls. Polymorphisms were detected in exon 4 (rs396991) and exon 3 (rs10127939) in the whole exom analysis of FcγRIIIA gene by new generation sequencing method. In addition, a possible chimeric region (exon 1 and exon 2 and exon 1 intron region) similar to FcγRIIIA and FcγRIIIB was determined by sequencing. In conclusion, in this study, patients with FNKD were evaluated with advanced functional and genetic analyzes. Differences were found in terms of NK number and functions. FcγRIIIA gene polymorphisms were identified related to the population of Turkey is made in the first evaluation will be done in the field of NK cell research is expected to shed light on the new study.
Natural killer cells (NK) are cells that play an important role in defending the organism against infections and cancer. NK cells have two important functions in the organism. The first is the direct cytotoxic effect against microorganisms that cause infection, Secondly, it is an antibodydependent cellular cytotoxicity (ADCC) that is mediated by the FcγRIIIA gene product. Considering the important role of NK cells, disorder(s) in the NK cell number or function cause disease. This disease originating from NK cells is called NK cell deficiency / disorder (NKD). From this point, NK cell subgroups, NK cell cytotoxicity and FcγRIIIA gene mutation(s) were investigated. The aim of this study was to evaluate the relationship between clinical, laboratory and functional analysis and FcγRIIIA gene mutation(s). The study group consisted of 10 patients who were admitted to Necmettin Erbakan University Meram Medical Faculty Pediatric Allergy and Immunology Policlinic for the first time and previously monitored and consisting of 7 healthy individuals were included in the control group. All patients and controls were evaluated for the determination of peripheral blood lymphocyte ratios and NK cell subgroups and cytotoxicity tests were performed to evaluate the functions of NK cells. FcγRIIIA gene mutations were investigated by next generation sequencing method. Three different NK cell subgroups, CD56brightCD16neg, CD56brightCD16int and CD56dimCD16hi were identified in the NK cell subgroup analysis evaluated by peripheral blood flow cytometry. No statistically significant difference was found in the ratio of CD56brightCD16neg cells when NK cell subsets of the patients were compared with the control group. CD56brightCD16int and CD56dimCD16hi cell ratios were found to be significantly lower in patients compared to the control group. NK cell cytotoxicity was evaluated by K562 lysis dependent method. As a result of NK cell cytotoxicity analysis, proportional decrease of K562 amount between patient and controls was found to be statistically significant (p<0,001). It was found that the rate of K562 cell reduction in the patients decreased statistically less than the controls. Polymorphisms were detected in exon 4 (rs396991) and exon 3 (rs10127939) in the whole exom analysis of FcγRIIIA gene by new generation sequencing method. In addition, a possible chimeric region (exon 1 and exon 2 and exon 1 intron region) similar to FcγRIIIA and FcγRIIIB was determined by sequencing. In conclusion, in this study, patients with FNKD were evaluated with advanced functional and genetic analyzes. Differences were found in terms of NK number and functions. FcγRIIIA gene polymorphisms were identified related to the population of Turkey is made in the first evaluation will be done in the field of NK cell research is expected to shed light on the new study.
Açıklama
Doktora Tezi
Anahtar Kelimeler
FcγRIIIA, NK, NK Alt Grup, Sitotoksisite, NK Subgroup, Cytotoxicity
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Karaselek, M. A. (2020). Doğal öldürücü (NK) hücre düşüklüğü olan hastalarda, NK Hücre Sitotoksitelerinin ve FcγRIIIA gen mutasyonlarının araştırılması. (Yayımlanmamış yüksek lisans tezi). Necmettin Erbakan Üniversitesi, Sağlık Bilimleri Enstitüsü Tıbbi Biyoloji Anabilim Dalı, Konya.
