Candan ÖZOĞUL, Deniz ERDOĞAN, Çiğdem ELMAS, S. Gülşen GİRAY

Puberteye kadar farklı yaş guruplarında çeşitli cd moleküllerinin timustaki tutumlarının immunohistokimyasal olarak belirlenmesi

Amaç: Bu çalışmada doğumdan puberteye kadar olan evrede, insan timusunda, CD3, CD4 ve CD8 antijenlerinin immünohistokimyasal dağılımları ve yaşa bağlı timik yapısal değişimlerin araştırılması amaçlandı. Gereç ve Yöntem: Timus doku örnekleri, Gazi Üniversitesi Tıp Fakültesi Hastanesi Kalp Damar Cerrahisi Anabilim Dalında çeşitli nedenlerle ameliyat olan ve yaşlan 11 aylıktan, 11 yaşma kadar değişen 8 hastadan elde edildi. Farklı yaş gruplarından alınan timus dokularında, CD3, CD4 ve CD8 moleküllerinin dağılımlarını tanımlamak için, dokuların tamamına parafın, bir grubuna da dondurma-kesit immünohistokimyası uygulandı. Bulgular: Bütün yaş gruplarında, timusun korteks ve medulla bölgelerinin timositlerle dolu olduğu ve timositlerde belirgin bir apopitosiz gözlenmemiştir. İmmünohistokimyasal bulgularda, erken dönemlerde, korteks lenfositlerindeki tutulumun yoğun membranöz ve sitoplazmik olduğu, buna karşın ilerleyen dönemlerde bu tutulumun kuvvetliden ortaya değiştiği izlendi. Ayrıca, medulladaki reaktif hücre sayısının ise CD3'e oranla oldukça az olduğunu saptandı. CD8 antikoru ile boyanan grupların, korteksindeki timositlerde yaygın membranöz tutulum saptandı. Sonuç: Çalışmanın sonucunda CD4 ve CD8'in benzer dağılım gösterdiği saptandı. Reaktivite korteks timositlerinin büyük çoğunluğunu, meduller timositlerin ise küçük bir grubunu, kuvvetli membranöz ve orta dereceli sitoplazmik olarak olarak kapsamaktaydı. CD3 reaksiyonu ise korteks ve medulla timositlerinde yaklaşık benzer dağılım gösterdi. Bulgular timusdaki T lenfosit farklanması aşamalarıyla uyumlu olarak değerlendirildi. Ekspresyon açısından yaşa bağlı önemli değişim saptanmadı. Timusun yaşa bağlı olarak involusyona uğramasına karşın timositlerin işlevlerini aynı etkinlikte sürdürdükleri kanısına varıldı.

Immunohistochemical investigation of various cd molecules on thymus form birth to puberty in difeerent age groups

Purpose: Immunohistochemical expression of CD3, CD4, and CD8 antigens in the thymus from birth to puberty was investigated in this study. We analyzed the histologic changes that occur in the thymus with age and the distrubution of cells that react with these antigens. Methods: Tissue samples were obtained from patients of Gazi University Faculty of Medicine, Department of Cardivascular Surgery. Age of the patients ranged from 11 months to 11 years old. Three of them were boys and five of them were girls. Both parraffin and cryo sections were taken and immunostaining was performed. Results: Thymocytes were seen in the cortex and medulla of all age groups. Definite apoptosis was not seen in the thymocytes. Immunohistochemieal findings show that although strongly positive membranous and cytoplasmic staining was seen in the cortex thymocytes of early age groups, this staining seems to change from strongly positive to mild with age in thymus samples. The most positive reaction was seen with CD3 in the medulla of all age groups. CD8 immunoreactivity in the cortex was Seen mostly in membranous staining. Conclusion: We observed that CD4 and CD8 had similar immunoreactions in the thymus. Reactive thymocytes include most of the cortex thymocytes and small amounts of medullar thymoctes. This reactivity was strongly membraneous and mild cytoplasmic' In addition, CD3 reactivity was similar in the cortex and medulla of all age groups. Findings were well-matched with the changing structure of the aging thymus. Major changes were not seen in different age groups. We estimated that the thymus involutes with age but thymocytes work with the same activity and efficiency.

PDF

___

1. Fagoaga OR, Yellon SM, Nehlsen-Cannarella SL: Maturation of lymphocyte immunophenotypes and memory T helper cell differentiation during development in mice, Dev Immunol. 2000; 8: 47-60.
2. Haeryfar SM, Berczi I: The thymus and acute phase response, Cellular and Molecular Biology, 2001; 47: 145-156.
3. Pearse G: Normal structure, function and histology of the thymus. Toxicologic Pathology, 2006, 34: 504-514.
4. Bosselut R, Feıgenbaum L, Sharrow SO, Singer A: Strength of signaling by CD4 and CD8 coreceptor tails determines the number but not the lineage direction of positively selected thymocytes, Immunity. 2001; 14: 483-94.
5. Barbier E, Cazenav PA, Grassi F: A role for CD8 in limiting degeneracy of thymocyte selection, Cell Mol Biol. 2001; 47: 129-133.
6. Cho EY, Bae Y, Lee DS, Park SH: T-T interaction during thymic selection, Cellular and Molecular Biology, 2001; 47: 135-143.
7. Tatsumi Y, Kumanogoh A, Saitoh M, Mizushima Y, Kimura K, Suziki S, Yağı H, Horiuchi A, Ogata M, Hamaoka T: Differentiation of thymocytes from CD3-CD4CD8- through CD3-CD4-CD8+ into more mature stages induced by a thymic stromal cell clone, Proc Natl Acad Sci. 1990; 87: 2750-2754.
8. Ward JM, Erexson CR, Faucette LJ, Foley JF, Dijkstra C, Cattoreeti G: Immunohistochemical markers for the rodent immune system. Toxicologic Pathology 2006 34: 6,16-630.
9. Havran WL, Poenie M, Kimura J, Tsien R, Weiss A, Allison JP: Expression and function of the CD3 -antigen receptor on murine CD4+8+ thymocytes, Nature, 1987; 330: 170-173.
10. Ramos-Vara JA, Miller MA, Lopez E, Prats N, Brevik L: Reactivity of polyclonal human CD3 antiserum in lymphoid tissues of cattle, sheep, goats, rats and mice, Am J. Vet. Res. 1994; 55: 63-66.
11. Yamazaki H, Tateyama H, Asai K, Fukai I, Fujii Y, Tada T, Eimoto T: Glia maturation factor-b is produced by thymoma and may promote intrarumoral T-cell differentiation. Histopathology 2005, 47, 292-302.
12. Gilfillan S, Shen X, Konig R: Selection and function of CD4+ lymphocytes in transgenic mice expressing mutant MHC class II molecules deficient in their interaction with CD4, J Immunol. 1998; 161:6629-6637.
13. Chan S, Margarida CN, Dierich A, Benoist C, Mathis D: Visualization of CD4/CD8 T cell commitment, J Exp Med 1998; 12: 2321-2333.
14. Maroto R, Shen X, Konig R.: Requirement for efficient interactions between CD4 and MHC class II molecules for survival of resting CD4+ T lymphocytes in vivo and for activation-induced cell death, J Immunol. 1999: 162: 5973-5980.
15. Hue S, Monteiro RC, Berrih-Aknin S, Caillat-Zucman S: Potential Role of NKG2D/MHC Class I-Related Chain A Interaction in Intrathymic Maturation of Single-Positive CD8 T Celisi. The Journal of Immunology 2003; 171-1909-1917.
16. MacDonald HR, Budd RC, Howe RC: A CD3- subset of CD4-8+ thymocytes: a rapidly cycling intermediate in the generation of CD4+8+ cells, Eur J Immunol. 1988; 18: 519-523.