Content and location of lymphocyte subpopulations with markers CD4+, CD8+ and CD20+ in the esophageal tonsil of chickens and the Meckel diverticulum of ducks

  • V. Т. Khomich National University of Life and Environmental Sciences of Ukraine
  • N. V. Dyshliuk National University of Life and Environmental Sciences of Ukraine
  • T. A. Mazurkevych National University of Life and Environmental Sciences of Ukraine
  • S. V. Guralskа Polissia National University
  • S. І. Usenko National University of Life and Environmental Sciences of Ukraine
Keywords: immunohistochemistry; lymphocyte markers; humoral immunity; cellular immunity; foreign antigens


Immune formations of birds' digestive organs, including the esophageal tonsil and Meckel’s diverticulum, protect the body from foreign antigens that enter the body with food and water and play an important role in maintaining the genetic constancy of its internal environment. This unique property of the immune system is formed during ontogenesis and is associated with maintaining the selection of lymphocyte clones that are able to respond to foreign antigens and carry out a specific immune response of two types: humoral and cellular. This article presents the results of a study of T- and B-lymphocyte subpopulations of the esophageal tonsil of Shever 579 cross chickens at the age of 25, 180 and 300 days, the Meckel diverticulum of the Blagovarsky cross ducks at the age of 30, 150 and 180 days. Immunohistochemical and statistical research methods were used to determine the localization and quantitative parameters of cell populations of lymphocytes (CD4+, CD8+, CD20+) using monoclonal antibodies and the DAKO EnVision FLEX+ imaging system (Dako Cytomation, Denmark). Separate subpopulations of T-lymphocytes (CD4+ - helpers, CD8+ -cytotoxic / T-suppressors) and mature B-lymphocytes (CD20+) were found in the esophageal tonsil and Meckel diverticulum of birds. Their presence confirms that antigen-independent proliferation and differentiation of lymphocytes into effector cells occur in the immune formations of the digestive system. The lymphoid tissue of these formations is represented mainly by a well-defined diffuse form and nodules with light centers (secondary). In the esophageal tonsil of chickens, these structures are located in the tunica mucosa and tela submucosa, and in the Meckel diverticulum of ducks – also in the tunica muscularis. The content of lymphocytes with these markers predominates in diffuse lymphoid tissue compared to that in secondary lymphoid nodules. In the diffuse lymphoid tissue of the esophageal tonsil, lymphocytes are located mainly near the adenomeres and excretory ducts of the esophageal glands, blood vessels, and under the surface epithelium, and in Meckel’s diverticulum – around the crypts, in their epithelium and in the epithelium of the villi. They are also found in the light centers of lymphoid nodules and on their periphery. The indices of the content of lymphocytes with the indicated markers in the esophageal tonsil and Meckel diverticulum which we determined were associated with age characteristics of the poultry in the postnatal period of ontogenesis. According to our observations, the content of CD20+ lymphocytes was the highest, while the populations of CD4+ and CD8+ lymphocytes were much smaller. This indicates an increase in the activity and predominance of the humoral immunity over the cellular one. The content of CD20+ lymphocytes was highest in birds at the age of 180 days, that is, during their sexual maturity. The data presented in the work can be used by morphologists researching the organs of the immune system, immunologists, poultry specialists involved in breeding, using and raising poultry and in educational work.


Ahluwalia, B., Magnusson, M. K., & Öhman L. (2017). Mucosal immune system of the gastrointestinal tract: Maintaining balance between the good and the bad. Scandinavian Journal of Gastroenterology, 52(11), 1185–1193.

Alitheen, N. B., McClure, S., & McCullagh, P. (2010). B-cell development: One problem, multiple solutions. Immunology and Cell Biology, 88(4), 445–450.

Avtandilov, G. G. (1990). Medicinskaja morfometrija [Medical morphometry]. Medicina, Moscow (in Russian).

Besoluk, К., & Eken, E. (2001). The arterial supply of Meckel’s diverticulum in geese (Anser anser domesticus). Journal Veterinary Medical Science, 63(12), 1343–1345.

Branton, S. L., Lott, B. D., Morgan, G. W., & Deaton, J. W. (1988). Position of Meckel’s diverticulum in broiler-type chichens. Poultry Science, 67(4), 677–679.

Casteleyn, D., Doom, M., Van den Broeck, W., Simoens, P., & Cornillie, P. (2010). Locations of gut associated lymphoid tissue in the 3-month-old chicken: A review. Avian Pathology, 39(3), 143–150.

Cesta, M. F. (2006). Normal structure, function, and histology of mucosa-associated lymphoid tissue. Toxicologic Pathology, 34, 599–608.

Ciriaco, E., Perez Pinera, P., Díaz-Esnal, B., & Laurà, R. (2003). Age-related changes in the avian primary lymphoid organs (thymus and bursa of Fabricius). Microscopy Research and Technique, 62(6), 482‒487.

Davison, F. (2014). The importance of the avian immune system and its unique features. In: Schat, K. A., Kaspers, B., & Kaiser, P. (Eds). Avian immunology. Academic Press, London. Pp. 1–9.

Debard, N., Sierro, F., Browning, J., & Kraehenbuhl, J.-P. (2001). Effect of mature lymphocytes and lymphotoxin on the development of the follicle-associated epithelium and M cells in mouse peyer’s patches. Gastroenterology, 120(5), 1173–1182.

Del Moral, G. M., Fonfría, J., Varas, A., Jiménez, E., Moreno, J., & Zapata, A. G. (1998). Appearance and development of lymphoid cells in the chicken caecal tonsil. The Anatomical Record, 250, 182–189.

Dishluk, N. V., & Orlova, A. V. (2017). Osoblyvosti budovy stravokhodu ta yoho imunnykh utvoren’ perepeliv [Structural features of esophagus and its immune formations in quails]. Scientific Messenger of Lviv National University of Veterinary Medicine and Biotechnologies named after S. Z. Gzhytskyj, 77(19), 3–6 (in Ukrainian).

Donmez, H. H., Eken, E., Besoluk, K., & Sur, E. (2012). The histological characteristics and localization of ACP and ANAE positive lymphocytes in the oesophageal tonsil of the duck (Anas platyrhynchos). Avian Biology Research, 5(1), 11–15.

Fellah, J. S., Jaffredo, T., Nagy, N., & Dunon, D. (2014). Development of the avian immune system. In: Schat, K. A., Kaspers, B., & Kaiser, P. (Eds.). Avian immunolоgy. Academic Press, London. Pp. 45‒63.

Göbel, T. W., Kaspers, B., & Stangassinger, M. (2001). NK and T cells constitute two major, functionally distinct intestinal epithelial lymphocyte subsets in the chicken. International Immunology, 13(6), 757‒762.

Gofur, M. R. (2020). Meckel’s diverticulum in animals and birds: An immuno-pathoclinical perspective. Bangladesh Journal of Veterinary Medicine, 18(1), 1–12.

Goralsky, L. P., Khomich, V. T., & Kononsky, O. I. (2015). Osnovy gistologichnoji tekhniky i morfofynctsionalni doslidzhennia u normi ta pry patologiji [Basics of hіstologіcal technіque and morphofunctіonal methods of researches in the norm and pathology]. Polissya, Zhytomyr (in Ukrainian).

Guralska, S. V. (2017). Immunohistochemistry of the organs of hemopoiesis and immunogenesis of chickens at infectious bronchitis. Veterinary Biotechnology, 31, 50–58.

Havrylina, O. H., & Evert, V. V. (2016). Metodychni osoblyvosti zastosuvannya imunohistokhimichnoyi diahnostyky tsyrkovirusnoyi infektsiyi svyney [Methodological features of application of immunohistochemical diagnostics of circovirus infection of pigs]. Problems of Zooengineering and Veterinary Medicine, 32(2), 294–301 (in Ukrainian).

Jaffe, E. S., Harris, N. L., Stein, H., & Vardiman, J. W. (2001). Pathology and genetics. Tumors of Haematopoietic and Lymphoid Tissues. IARC Press, Lyon.

Jeurissen, S. H. M., Janse, E. M., Koch, G., & De Boer, G. F. (1989). Postnatal development of mucosa-associated lymphoid tissues in chickens. Cell and Tissue Research, 258, 119–124.

Karadag, S. E., Altunay, H., Kurtdede, N., & Bakir, B. (2015). The structure of bursa of fabricius in the long-legged buzzard (Buteo rufinus): Histological and histochemical study. Actа Veterinaria, 65(4), 510–517.

Kaspers, B., & Göbel, T. W. F. (2016). The avian immune system. In: Ratcliffe, M. J. H. (Ed.). Encyclopedia of immunobiology. Elsevier Ltd. Vol. 1. Pp. 498–503.

Knowles, D. M. (2000). Neoplastic hematopathology. 2nd ed. Lippincott Williams & Wilkins, Philadelphia.

Kotsyumbas, I. Y., Zhyla, M. I., Shkodyak, N. V., Sobodosh, S. Y., & Martynyk, S. Y. (2014). Retseptory poverkhnevykh struktur imunokompetentnykh klityn u tvaryn. Suchasni metody yikh vyznachennya [Receptors of surface structures of immunocompetent cells in animals. Modern methods of their determination]. Scientific and Technical Bulletin of the Institute of Animal Biology and the State Research Control Institute of Veterinary Drugs and Feed Additives, 15(2), 349–355 (in Ukrainian).

Koutsos, E. A., & Klasing, K. C. (2014). Factors modulating the avian immune system. In: Schat, K. A., Kaspers, B., & Kaiser, P. (Eds.). Avian immunolоgy. Academic Press, London. Pp. 299–313.

Kovtun, M. F., & Kharchenko, L. P. (2005). Limfoidnyye obrazovaniya pishchevaritel’noy trubki ptits: Kharakteristika i biologicheskoye znacheniye [Lymphoid formations of the digestive tube of birds: Characteristics and biological significance]. Vestnik Zoologii, 39(6), 51–60 (in Ukrainian).

Laky, K., Fleischacker, C., & Fowlkes, B. J. (2006). TCR and Notch signalingin CD4 and CD8 T-cell develjpment. Immunological Reviews, 209, 274–283.

Logvinova, V. V., Oliyar, A. V., & Lieshchova, M. A. (2020). Formuvannya imunnykh struktur tonkoyji kyshky muskusnykh kachok (Cairina moschata) [Formation of immune structures in small intestine of Muscovy ducks (Cairina moschata)]. Theoretical and Applied Veterinary Medicine, 8(1), 50–55.

Macardle, P. J., & Nicholson, I. C. (2002). CD20. Journal of Biological Regulators and Homeostatic Agents, 16(2), 136–138.

Nagy, N., Igyártó, B., Magyar, A., Gazdag, E., Palya, V., & Olah, L. (2005). Oesophageal tonsil of the chicken. Acta Veterinaria Hungarica, 53(2), 173–188.

Nakamura, K., Yube, A., Miyatake, J. C., & Cambier, M. (2003). Hirashima involvement of CD4 D3-D4 membrane proximal extracellular domain for the inhibitory effect of oxidative stress on activation-induced CD4 down-regulation and its possible role for T cell activation. Molecular Immunology, 39, 909–921.

Olah, I., & Glick, B. (1984). Meckel’s diverticulum. I. Extramedullary myelopoiesis in the yolk sac of hatched chickens (Gallus domesticus). The Anatomical Record, 208, 243–252.

Oláh, I., Nagy, N., & Vervelde, L. (2013). Structure of the аvian lymphoid system. In: Schat, K. A., Kaspers, B., & Kaiser, P. (Eds.). Avian immunology. Academic Press, London. Pp. 11–44.

Panikar, I. I. (2015). Imunohistokhimichna kharakterystyka subpopulyatsiy limfotsytiv orhaniv imunnoyi systemy svyney na rannikh etapakh postnatal’noho periodu ontohenezu [Immunohistochemical characteristics of subpopulations of lymphocytes of the immune system of pigs in the early stages of the postnatal period of ontogenesis]. Problems of Zooengineering and Veterinary Medicine, 30(2), 426–432 (in Ukrainian).

Polegenka, M. A. (2019). Analiz suchasnoho stanu vyrobnytstva produksiyi ptakhivnytstva v Ukrajini [Analysis of the current state of production of poultry products in Ukraine]. Ekonomika i Derzhava, 3, 137–143 (in Ukrainian).

Rahman, M. L., Islam, M. R., Masuduzzaman, M., & Khan, M. Z. I. (2003). Lymphoid tissues in the digestive tract of deshi chicken (Gallus domesticus) in Bangladesh. Pakistan Journal of Biological Sciences, 6, 1145–1150.

Samour, J. (2015). Avian medicine. 3rd ed. Mosby Ltd., Город.

Schepers, K., Arens, R., & Schumacher, T. N. M. (2005). Dissection of cytotoxic and helper T cell responses. Cellular and Molecular Life Sciences, 62(23), 2695–2710.

Sharma, J. M. (1991). Overview of the avian immune system. Veterinary Immunology and Immunopathology, 30(1), 13–17.

Smith, P., MacDonald, T., & Blumberg, T. (2012). Principles of mucosal immunelogy. Garland Science, New York.

Walport, M., Murphy, K., Janeway, C., & Travers, P. J. (2008). Janeway’s Immunobiology. 7th ed. Garland Science, New York.

Yamanaka, T., Straumfors, A., Morton, H. C., Fausa, O., Brandtzaeg, P., & Farstad, I. N. (2001). M cell pockets of human Peyer’s patches are specialized extensions of germinal centers. European Journal of Immunology, 31, 107–117.

Yasuda, M., Kajiwara, E., & Ekino, S. (2003). Immunobiology of chicken germinal center: I. Changes in surface Ig class expression in the chicken splenic germinal center after antigenic stimulation. Developmental and Comparative Immunology, 27(2), 159–166. 

How to Cite
KhomichV. Т., Dyshliuk, N. V., Mazurkevych, T. A., GuralskаS. V., & UsenkoS. І. (2021). Content and location of lymphocyte subpopulations with markers CD4+, CD8+ and CD20+ in the esophageal tonsil of chickens and the Meckel diverticulum of ducks . Regulatory Mechanisms in Biosystems, 12(3), 396-402.