Investigation of Histochemical and Histometric Distribution of Lyphocytes in Thymus and Blood Tissues Due to Rats Age

Year 2018, Volume: 4 Issue: 2, 900 - 920, 15.08.2018

Alper Yalçın , Mehmet Kanter

https://doi.org/10.30569/adiyamansaglik.441354

Abstract

Objective: This study was performed to investigate the histochemical and
histometric distribution of lymphocytes in thymus and blood tissues of the rats
depending on their different age stages

Material and Method:  In the study, 60 Wistar
albino rats were used. Thymus and blood tissue samples were taken following
decapitation. Thymus tissues were firstly placed in formal-sucrose and then
Holtz solutions for fixation. Blood samples were put into the heparinized
tubes. To determine the alpha naptyhl acetate esterase  (ANAE) positive lymphocytes from the both
tissues, thymus tissue sections cut into 10 µ thickness by cryostat microtome
and smears made from blood samples were stained in the incubation solutions for
5 minutes and  3  hours 
at  pH  5.8, 
respectively.

.Results: When the ANAE  (+)  positive
lymphocytes existence in cortical and medullar parts of the thymic tissue were
evaluated for each stage statistically; the most numbers in puberty and the
least in adult stage were found respectively whereas the pre-puberty stage were
found to have much more positive lymphocytes compared with the adult stage. In
the blood tissue samples, the numbers of ANAE positive lymphocytes were found
to be very few in pre-puberty stage, the most in the puberty stage; however,
the numbers of the cells in the adult stage were fewer than puberty but more
than pre-puberty stages. The ANAE negative lymphocytes in the blood tissue
samples were regarded as B lymphocytes

Conclusion: While the rate of lymphocytes in pre-puberty and puberty stages
in blood samples paralleled with their distribution in the thymic tissue, the
rate of positive lymphocytes in peripheral blood was much more higher than
thymus tissue only in the adult stage, which it could be postulated that
abundant numbers might have been because of thymus involution or constant
proliferation of the memory T cells in periphery.

Keywords

Age, ANAE, histochemistry, lymphocytes, rat

Sıçanlarda Yaşa Bağlı Olarak Timus ve Periferal Kan Dokusunda bulunan Lenfositlerin Histokimyasal ve Histometrik İncelenmesi

Abstract

Amaç: Bu çalışma, sıçanlarda yaşa bağlı olarak
timus ve periferal kan dokusunda bulunan lenfositlerin histokimyasal ve histometrik
incelenmesi amacıyla yapıldı.

Gereç ve Yöntem: Çalışmada 60 Wistar Albino sıçan kullanıldı.  Farklı yaş grubundaki sıçanlar dekapite
edilerek timus ve kan dokuları uzaklaştırıldı. Timus dokuları sırasıyla
formol-sukroz ve Holtz solüsyonunda tespit edildi. Kan dokuları ise heparinli
tüplere alındı. Alfa naftil asetat esteraz 
(ANAE) pozitif lenfositleri belirlemek amacıyla kriyostatta timus dokusundan
alınan kesitler, pH 5.8'de 5 dakika, kan dokusundan yapılan frotiler ise pH 5.8
'de 3 saat süre ile inkübasyon solüsyonunda boyandı.

Bulgular: Timusta her dönem için korteks ve
medullada bulunan toplam ANAE (+) lenfosit dağılımı istatistiksel olarak
değerlendirildiğinde en fazla puberte, en az ergin dönemde bulunduğu;
prepuberte döneminde ise ergin döneme göre bu sayının anlamlı bir şekilde fazla
oldugu saptandı. Kan dokusunda  ise ANAE
pozitif lenfosit sayılarının prepuberte döneminde oldukça az, puberte  dönerninde 
en fazla, ergin dönemde ise puberteye göre az ancak prepuberte dönemine  göre 
fazla olduğu saptandı. Kan dokusunda ANAE negatif lenfositler B lenfosit
olarak değerlendirildi.

Sonuç: Kan ömeklerinde ANAE (+) lenfositlerin
oranı prepuberte ve puberte dönernlerinde timustaki dağılımla paralellik
gösterirken, sadece ergin dönemde periferal kandaki pozitiflik oranının timusa
göre oldukça yüksek çıkması timustaki involusyona bağlanabileceği gibi,
periferdeki bellek T hücrelerinin sürekli olarak proliferasyonu sonucu da bu
artışın olabilecegi kanaatini taşımaktayız.

Keywords

ANAE, histokimya, lenfosit, sıçan, yaş

References

• 1. Karin E. de Visser, Alexandra Eichten, Lisa M. Paradoxical roles of the immune system during cancer development. Coussens Nature Reviews Cancer volume 2006;6:24–37.
• 2. Kay BMM. The thymus : Clock of immunologic aging?. J Invest Dermatol 1979;73,29-38.
• 3. Gillis S, Kozak R, Durante M. Immunological studies of aging Decreased production of and response to T cell growth factor by lymphocytes from aged humans. J.Clin. Invest 1981;67,937.
• 4. Utsuyama M, Hirokawa K, Kurashima C et all. Differential age-change of CD4 + CD45RA + and CD4 + CD29+ T cells subsets in human peripheral blood. Mech Ageing Dev 1992;10:57–68.
• 5. Hirokawa K, Utsuyama M, Makinodan T. Immunity and ageing. In: Principles and Practice of Geriatric Medicine. 4. Pathy MSJ, Sinclair AJ, Morley JE, editor. John Wiley and Sons, 2006:19–36.
• 6. Scholz JL, Diaz A, Riley RL et all. Curr Opin Immunol: A comparative review of aging and B cell function in mice and humans. Epub 2013;4:504-510.
• 7. Fabris N. Immunulogy and Aging. Experientia, 1981;37:1041-1043
• 8. Sigal HL, Ron Y. Immunology Inflammation, McGraw-Hill, Inc, USA 1994.
• 9. Nociari MM, Telford Wand Russo C. Postthymic development of CD28 CD8+ T cellsubset: Age-associated expansion and shift from memory to naive phenotype. J Immunol 1999; 162:6,3327-3335.
• 10. Hirokawa K. Understanding the mechanism of the age-irel;tted decline in immune function. Nutr Rev 1992; 50(12):361-366.
• 11. Caruso C, Lio D, Cavallone L et all. Aging, longevity, inflammation and cancer. Ann NY Acad Sci 2004;1028:1-13.
• 12. Hurme M. Immunity in the elderly. Article in Finnish, Duodecim 2013;18:1878-1885.
• 13. Higgy EK, Bums FG, Hayhoe JGF. Discrimination of B, T and null lymphocytes by esrerase cytochemistry. Scand J Haematol 1977;18:437-448.
• 14. Basso G, Cocito GM, Semenzato G et all. Cytochemical study of thymocytes and T lymphocytes. Br J Haematol 1980; 44: 577- 582.
• 1. 15.Yang TJ, Patricia JA, Williams LF. Acid alpha naphthyl acetate esterase: Presence of activity in bovine and human T and B lymphocytes. Immunology 1979;38:85-92.
• 15. Monahan AR, Dvorak FH and Dvorak MA. Ultrastructural localization of nonspesific esterase activity in guinea pig and human monocytes, macrophages and lymphocytes, Blood, 1981;58: 6.
• 16. Li CY, Lam W and Yam LT. Esrerases in leucocytes, J Histochem Cytochem, 1973;21:1-12.
• 17. Mueller G, Buerki H, Keller HWM and Cottier H . Nonspesific acid esterase activity. A criterion for differentiation ofT and B lymphocytes in mouse lymph nodes, Eur J Immunol. 1975;5,:270-274.
• 18. Hovmark A. Acid a. naphthyl acetate esterase staining ofT lymphocytes in human skin, Acta Derm Venereal, 1977;57:497-502.
• 19. Knowles MD and Holck S. Tissue localization ofT-lymphocytes by the histochemical demonstration of acid alfa napthyl acetate esterase, Lab Invest, 1978;39(1):70-76.
• 20. Palestra G, Valente G, Micca BV et all. Detection and distribution of alfa-naphthyl acetate esterase activity in thymocytes of normal myasthenic thymus and thymoma. VirchowsArch B C ell Pathol 1980; 35: 33-43.
• 21. Özdamar, K. Spss ile Biyoistatistik, Yenilenmiş 5. baskı, Ankara: Kazan Kitapevi, 2003.
• 22. Kulenkumph J, Janossy G, Greaves MF. Acid esterase in human lymphoid cells and leukaemic blasts: A marker for T lymphocytes. Br J Haematol 1977;36:231-240.
• 23. Knowles MD, Halper PJ. Human medullary and cortical thymocytes are distinguishable according to the presence or absence of cytochemically demonstrable acid a. naphthyl acetate esterase (ANAE) activity. J Immunol 1980; 6:2823-2825.
• 24. Bianchi ATJ, Zwart RJ, Jeurissen SHM et all. Development of the B and T cell compartments in porcine lymphoid organs from birth to adult life: An immunohistological approach. Vet Immunol Jmmunopathol 1992;33:201-221.
• 25. Djilali S, Dacosta B, Kessler JL et all. Preparation and characterization of a monoclonal antibody against bovine CDS lymphocyte surface antigen. Comp Immun Microhiol Infect Dis 1991;3:257-263.
• 26. Knowles MD, Halper PJ, Machin AG et all. Acid a. naphthyl acetate esterase activity in human neoplastic lymphoid cells. Am J Pathol 1979;1:257-270.
• 27. Demir R. İnsanın Gelişimi implantasyon Biyolojisi. 1. Baskı, Ankara: Palme Yayıncılık, 1995.
• 28. Kato S, Kurihara K. Nonspesific esterase activity in monkey lymphocytes: Study of distribution in lymphocyte subpopulation,. Experientia 1981 37:1026-1027.
• 29. Pinkus SG, Hargreaves KH, McLeod AJ et all. A naphthyl acetate esterase activity: A cytochemical marker for T lymphocytes. Am JPathol 1979; 97: 17-42.
• 30. Ranki A, Totterman TH, Hayry P. Identification of mouse T and B lymphocytes from cytocentrifuged cell smears. Clin Exp Immunol 1976; 26: 632.
• 31. Kajikawa 0, Koyama H, Yoshikawa T et all. Use of apha-naphtyl acetate esterase staining to identify T lymphocytes in cattle. Am J VetRes 1982; 8: 1549-1552.
• 32. Yörük M, Aştı NR, Kurtdere N et all. Light and electron microscopic studies on alpha naphtyl acetate esterase activity of the peripheral blood T lymphocytes in Van Cats. Anat Histol Embryol 1998; 27:289-292.
• 33. Wada N, Prieur AM, Griscelli C. Nonspesific alpha naphthyl acetate esterase activity of T lymphocytes: Study in healthy newborn and children, in immune deficiencies and juvenile rheumatoid arthritis. Pediatr Res 1981; 15: 1266- 1270.
• 34. Aştı NR, Çelik İ, Kadak R ve ark. Farklı yaşlardaki sığırların kan T lenfosit oranlarında görülen değişiklikler. Hayvancılık Araştırma Dergisi 1993;3:2.
• 35. Otis LL, Larsen AE. Identification of T lymphocytes by a histochemical stain for alpha naphthyl acetate esterase activity. Am J Med Technol 1982;5: 367-370.
• 36. Bergroth V, Konttmen YT, Rettamo S. A method for the identification ofhuman peripheral blood T lymphocytes by seguentialtmmunogold and esterase double staining. J Histochem Cytochem 1983; 6: 837-839.
• 37. Windmill FK, Meade BJ, Lee KWV. Effect of prepubertal gonadectomy and sex steroid treatment on the growth and lymphocyte populations of the rat thymus. Reprod Fertil Dev 1993; 5: 73-81.
• 38. Paul PS, Senogl_es DR, Muscoplat CC et all. Enumaration ofT cells, B cells and monocytes in the peripheral blood of normal and lymphocytic cattle. Clin Exp Immunol 1979; 35: 306-316.
• 39. Yang TJ, Patricia JA, Williams LF. Acid alpha naphthyl acetate esterase: Presence of activity in bovine and human T and B lymphocytes. Immunology 1979; 38: 85-92.
• 40. Çelik İ, Vural Ö, Dönmez H ve ark. Determination of percentages in peripheral blood and tissue localization of T-lymphocytes in fetal and adult lymphoid tissues of cattle by the histochemical demonstration of alpha­ naphtyl acetate esterase. Veterinarium 1993;1:10-17.
• 41. Klempau EA, Cooper LE. T-lymphocytes and B lymphocytes in AnuranAmphh1b1ans: T-lymphocytes proportions, distribution and ontogeny, as measured by E-rosetting, nylon wool adherence, postmetamorphic thymectomy and non spesific esterase staining. Dev Comp Immunol 1983;7:99-110.