Publication: Spontan ve idiyopatik rekürren düşük yapan kadınlarda natural killer (NK) T hücresi immünoglobulin ve müsin dome -3 (TİM-3) düzeylerinin araştırılması
Abstract
Amaç: Bozulmuş tolerans indüksiyonu, aşırı inflamasyon spontan-rekürren abortus gibi gebelik komplikasyonlarına yol açmaktadır. TİM-3 birçok fizyolojik ve patolojik süreçte temel immünolojik fonksiyona sahip yeni tanımlanmış molekül olup bağışıklık toleransının düzenlenmesinde rol oynamaktadır. Bu çalışmanın amacı, gebeliğin 1.Trimesterinde Spontan(SA) ve Reküren Abortus(RA) yapan gebeler ile 1.Trimester normal gebelik süreci devam eden gebe(SG) ve sağlıklı gebelik geçirmiş, sağlıklı doğum yapmış(S) kadınların venöz kanlarından izole edilen Periferal Kan Mononükleer Hücre(PKMH) fenotipleri arasındaki farka dayalı immünolojik reddin nedeninin araştırılmasıdır.Gereç-Yöntem: Genetik, endokrinolojik, anatomik ve çevresel faktörlerin dışlandığı idiopatik rekürren ve spontan abortus vakaları dahil edilmiştir. RA, SA, SG ve S deneklerinin venöz kanlarından PKMH izole edildi. Lenfositler(5x105) 48’lik platede Carboxyfluorescein succinimidyl ester (CFSE) boyalı-boyasız, Uyarımsız (US) ve IL- 4 Uyarımlı(IL4) ortamda 48 saat inkübe edildi. İnkübasyon sonrasında CFSE işaretli hücrelerin proliferasyonlarına, T regülatör hücre (Treg) düzeylerine, lenfosit apoptozu ve NK hücrelerinde Tim-3 ifadesi incelendi.xBulgular: Sonuçlara göre CD3-CD16+CD56+ periferik (p)NK hücreleri, US-SG PKMH kültürlerine göre US-SA(p<0.02) ve US-RA(p<0.008) kültürlerinde istatistiksel olarak anlamlı düşük bulundu.CD3-CD56+ pNK hücrelerinde TİM-3 düzeyi IL4-SA(p<0.002) ile IL4-RA(p<0,0001) PKMH kültürlerinde US-SA ile US- RA kültürlerine göre istatistiksel olarak anlamlı yüksek bulundu. US-SG grubuna göre US-SA grubunda CD4+CD25+FOXP3+ Treg hücreleri istatistiksel olarak anlamlı yüksek bulundu(p<0.007). IL-4 uyarımı ile IL4-SA grubuna göre US-SA grubunda CD4+CD25+FOXP3+ Treg hücreleri istatistiksel olarak anlamlı yüksek bulundu(p<0.01).Sonuçlar: RA, SA gebelerinde pNK TİM-3 düzeyi azalmıştır. Erken gebelikte, SA ve RA oluşumunu öngörmek için TİM-3 biyolojik belirteç olarak kullanılabilirliği önerilmektedir.
Aim:Altered induction tolerance and excessive inflammation can cause to complication in pregnancy such as spontaneous recurrent abortion.TIM-3 is newly determined term in various physiologic and patologic pathway which has basic immunologic function.It has a role in regulation of immune tolerance.The aim of the study is to establish reasons of immunological rejection by looking at Peripheral Blood Mononuclear Cell(PBMC) derived from pregnant’s venous blood who undergo Spontaneous(SA) and Recurrent abortion(RA) in their first-trimester also who in the healty pregnancy period and has had healthy pregnancy.Method:Idiopathic recurrent and spontaneous abortion cases which genetic, endocrinologic, anatomic,external factors were excluded,were included.PBMC’s were isolated from venous blood whom RA,SA,SG and S subjects.Lymphocytes(5x105) were incubated for 48 hours in 48 well plates as Carboxyfluorescin succinimidyl ester(CFSE) stained and not,un-stimulated(US) and IL-4 stimulated.Subsequently, cells proliferation who targeted with CFSE,level of T regulatory cells(Tregs), apoptosis in lymphocytes and TIM-3 expression in Natural Killers(NK) were examined.Result:According to the datas,CD3-CD16+CD56+ peripheral NK cells,PBMC culture in US-SG also US-SA(p<0.02) and US-RA(p<0.008) cultures were obtained statistically logical in abortion.Level of TIM-3 was found statistically higher at CD3- CD56+ pNK cells in IL4-SA(p<0.002) and IL4-RA(p<0,0001) PBMC cultures than US-SA and US-RA cultures.Due to US-SG group, CD4+CD25+FOXP3+Treg cells were found statistically logically high in US-SA group(p<0.007).Due to IL4 stimulated and IL4-SA group,CD4+CD25+FOXP3+Treg cells were found statistically logically high in US-SA group(p<0.01).xiiConclusion:Level of pNK TIM-3 were decreased in SA and RA pregnancies.So,TIM- 3 can be used as a biomarker to anticipate occurrence of SA and RA abortion in early- pregnancy.
Aim:Altered induction tolerance and excessive inflammation can cause to complication in pregnancy such as spontaneous recurrent abortion.TIM-3 is newly determined term in various physiologic and patologic pathway which has basic immunologic function.It has a role in regulation of immune tolerance.The aim of the study is to establish reasons of immunological rejection by looking at Peripheral Blood Mononuclear Cell(PBMC) derived from pregnant’s venous blood who undergo Spontaneous(SA) and Recurrent abortion(RA) in their first-trimester also who in the healty pregnancy period and has had healthy pregnancy.Method:Idiopathic recurrent and spontaneous abortion cases which genetic, endocrinologic, anatomic,external factors were excluded,were included.PBMC’s were isolated from venous blood whom RA,SA,SG and S subjects.Lymphocytes(5x105) were incubated for 48 hours in 48 well plates as Carboxyfluorescin succinimidyl ester(CFSE) stained and not,un-stimulated(US) and IL-4 stimulated.Subsequently, cells proliferation who targeted with CFSE,level of T regulatory cells(Tregs), apoptosis in lymphocytes and TIM-3 expression in Natural Killers(NK) were examined.Result:According to the datas,CD3-CD16+CD56+ peripheral NK cells,PBMC culture in US-SG also US-SA(p<0.02) and US-RA(p<0.008) cultures were obtained statistically logical in abortion.Level of TIM-3 was found statistically higher at CD3- CD56+ pNK cells in IL4-SA(p<0.002) and IL4-RA(p<0,0001) PBMC cultures than US-SA and US-RA cultures.Due to US-SG group, CD4+CD25+FOXP3+Treg cells were found statistically logically high in US-SA group(p<0.007).Due to IL4 stimulated and IL4-SA group,CD4+CD25+FOXP3+Treg cells were found statistically logically high in US-SA group(p<0.01).xiiConclusion:Level of pNK TIM-3 were decreased in SA and RA pregnancies.So,TIM- 3 can be used as a biomarker to anticipate occurrence of SA and RA abortion in early- pregnancy.