ISSN: 1301-2193 E-ISSN: 1308-9846
  • Turkish Journal of
    Endocrinology and Metabolism
ORIGINAL ARTICLE

Immunohistochemical Evidence of Apoptosis in Different Cells of Hashimoto's Thyroiditis and Graves' Disease Thyroids - Original Article
Immunohistochemical Evidence of Apoptosis in Different Cells of Hashimoto's Thyroiditis and Graves' Disease Thyroids - Original Article
Makale Dili: EN
ABSTRACT

Abstract
Objective:
Hashimoto’s (HT) and Graves’ (GD) diseases are thyroid autoimmune diseases that share similar pathogenic mechanisms. Apoptosis is often related to an autoimmune process and is present in HT thyroid follicular cells. We compared the apoptosis characteristics in these two thyroid diseases considering three thyroid cell types, namely lymphocytes, follicular cells and macrophages.
Material and Methods: Nineteen surgically removed thyroids coming from fourteen patients suffering from HT and five patients from GD, have been processed for immunohistochemistry. Apoptosis has been revealed using a terminal deoxynucleotidyl transferase - mediated dUTP nick end-labeling (TUNEL) and an anti active caspase-3 antibody respectively showing DNA fragmentation and activation of the last apoptosis effector. Bcl2, bax and p53 proteins were also immunohistochemically investigated.
Results: Lymphocytic infiltration, TUNEL positivity, caspase-3 activation, p53 and bax immunoreactivities were much more present in HT than in GD. Lymphoid cells reacted with anti bcl2 antibody only, with different patterns in HD and in GD. In GD, almost all the follicular cells were immunoreactive with anti bcl2 antibody. In HT, some follicular cells in the vicinity of lymphoid follicles were TUNEL, active caspase-3 and bax positive; on the contrary, far from lymphoid follicles, the follicular cells were bcl2 positive. Our main result showed the presence of numerous macrophages in HT thyroid follicular lumina, expressing strong bax and p53 immunoreactivities along with bcl2, TUNEL and active caspase-3 negativities.
Conclusion: Apoptosis is rare in GD thyroids and characteristic of HT thyroid follicular cells in the vicinity of lymphoid follicles. In HT, macrophages initiated apoptosis in follicular lumina and seemed either to stop apoptosis before its end or to commit suicide by a caspase-3 independent pathway without DNA fragmentation. Turk Jem 2007; 11: 73-8
Key words: Apoptosis, autoimmune thyroid diseases, TUNEL, caspase-3, bax, bcl2, p53

Özet
Amaç:
Hashimoto (HT) ve Graves (GH) hastalıkları tirodin otoimmun hastalıkları olup aynı patojenik mekanizmaları paylaşmaktadır. Apoptosis sıklıkla otoimmun olaylara bağlı olarak HT tiroid follikül hücrelerinde bulunmaktadır. Biz bu iki tiroid hastalığında üç tip tiroid hücresini lenfositler, follikül hücreleri ve makrofajları karşılaştırdık.
Gereç ve Yöntem: On-dokuz cerrahi olarak alınmış tiroid (14’ü HT, 5 hasta GH) immunohistokimyasal incelemeye alındı. Apoptosis “terminal deoxynucleotidyl transferase - mediated dUTP nick end-labeling” (TUNEL) ve anti aktif kaspaz 3 antikor kullanılarak  sırasıyla DNA fragmentasyonunu ve son apoptoz etkeni aktivasyonu bakılarak apoptosis değerlendirildi. Bcl2, bax ve p53 proteinleri de immunohistokimyasal olarak araştırıldı.
Bulgular: Lenfositik infiltrasyon TUNEL pozitifliği, kaspaz 3 aktivasyonu, p53 ve bax immunoreaktivitesi HT’da GH’na gore daha fazla bulundu. Lenfoid hücreler yalnızca  bcl2 antikoruyla HT ve GH’da farklı paternde reaksiyon gösterdi. GH’da hemen tüm folliküler anti bcl2 antikoru ile imunoreaktifdi. HT’da lenfoid follikül civarındaki bazı follikül hücreleri TUNEL, aktif kaspaz-3, ve bax pozitifdi. Buna karşın lenfoid folliküllerden uzak folliküler hücreler bcl2 pozitif bulundu. Ana bulgumuz olarak HT’da tiroid follikül lumeninde çok sayıda kuvvetli bax ve p53 immunoreaktivitesi ve bcl2, TUNEL ve aktif kaspaz 3 negatifliği gösteren çok sayıda makrofaj varlığı gözlendi.
Sonuç: Apoptosis GH tiroidinde nadir olup, HT için karakteristik olarak lenfoid folliküller çevresinde olur. HT’da makfofajlar apoptosisi follikül lümeninde başlatıyor ve apoptosisi DNA fragmantasyonu olmadan kaspaz-3’den bağımsız bir yolda durduruyor görünmektedir.  Turk Jem 2007; 11: 73-8
Anahtar kelimeler: Apoptosis, Otoimmun tiroid hastalıkları, TUNEL, caspase-3, bax, bcl2, p53

ÖZET

Abstract
Objective:
Hashimoto’s (HT) and Graves’ (GD) diseases are thyroid autoimmune diseases that share similar pathogenic mechanisms. Apoptosis is often related to an autoimmune process and is present in HT thyroid follicular cells. We compared the apoptosis characteristics in these two thyroid diseases considering three thyroid cell types, namely lymphocytes, follicular cells and macrophages.
Material and Methods: Nineteen surgically removed thyroids coming from fourteen patients suffering from HT and five patients from GD, have been processed for immunohistochemistry. Apoptosis has been revealed using a terminal deoxynucleotidyl transferase - mediated dUTP nick end-labeling (TUNEL) and an anti active caspase-3 antibody respectively showing DNA fragmentation and activation of the last apoptosis effector. Bcl2, bax and p53 proteins were also immunohistochemically investigated.
Results: Lymphocytic infiltration, TUNEL positivity, caspase-3 activation, p53 and bax immunoreactivities were much more present in HT than in GD. Lymphoid cells reacted with anti bcl2 antibody only, with different patterns in HD and in GD. In GD, almost all the follicular cells were immunoreactive with anti bcl2 antibody. In HT, some follicular cells in the vicinity of lymphoid follicles were TUNEL, active caspase-3 and bax positive; on the contrary, far from lymphoid follicles, the follicular cells were bcl2 positive. Our main result showed the presence of numerous macrophages in HT thyroid follicular lumina, expressing strong bax and p53 immunoreactivities along with bcl2, TUNEL and active caspase-3 negativities.
Conclusion: Apoptosis is rare in GD thyroids and characteristic of HT thyroid follicular cells in the vicinity of lymphoid follicles. In HT, macrophages initiated apoptosis in follicular lumina and seemed either to stop apoptosis before its end or to commit suicide by a caspase-3 independent pathway without DNA fragmentation. Turk Jem 2007; 11: 73-8
Key words: Apoptosis, autoimmune thyroid diseases, TUNEL, caspase-3, bax, bcl2, p53

Özet
Amaç:
Hashimoto (HT) ve Graves (GH) hastalıkları tirodin otoimmun hastalıkları olup aynı patojenik mekanizmaları paylaşmaktadır. Apoptosis sıklıkla otoimmun olaylara bağlı olarak HT tiroid follikül hücrelerinde bulunmaktadır. Biz bu iki tiroid hastalığında üç tip tiroid hücresini lenfositler, follikül hücreleri ve makrofajları karşılaştırdık.
Gereç ve Yöntem: On-dokuz cerrahi olarak alınmış tiroid (14’ü HT, 5 hasta GH) immunohistokimyasal incelemeye alındı. Apoptosis “terminal deoxynucleotidyl transferase - mediated dUTP nick end-labeling” (TUNEL) ve anti aktif kaspaz 3 antikor kullanılarak  sırasıyla DNA fragmentasyonunu ve son apoptoz etkeni aktivasyonu bakılarak apoptosis değerlendirildi. Bcl2, bax ve p53 proteinleri de immunohistokimyasal olarak araştırıldı.
Bulgular: Lenfositik infiltrasyon TUNEL pozitifliği, kaspaz 3 aktivasyonu, p53 ve bax immunoreaktivitesi HT’da GH’na gore daha fazla bulundu. Lenfoid hücreler yalnızca  bcl2 antikoruyla HT ve GH’da farklı paternde reaksiyon gösterdi. GH’da hemen tüm folliküler anti bcl2 antikoru ile imunoreaktifdi. HT’da lenfoid follikül civarındaki bazı follikül hücreleri TUNEL, aktif kaspaz-3, ve bax pozitifdi. Buna karşın lenfoid folliküllerden uzak folliküler hücreler bcl2 pozitif bulundu. Ana bulgumuz olarak HT’da tiroid follikül lumeninde çok sayıda kuvvetli bax ve p53 immunoreaktivitesi ve bcl2, TUNEL ve aktif kaspaz 3 negatifliği gösteren çok sayıda makrofaj varlığı gözlendi.
Sonuç: Apoptosis GH tiroidinde nadir olup, HT için karakteristik olarak lenfoid folliküller çevresinde olur. HT’da makfofajlar apoptosisi follikül lümeninde başlatıyor ve apoptosisi DNA fragmantasyonu olmadan kaspaz-3’den bağımsız bir yolda durduruyor görünmektedir.  Turk Jem 2007; 11: 73-8
Anahtar kelimeler: Apoptosis, Otoimmun tiroid hastalıkları, TUNEL, caspase-3, bax, bcl2, p53