1918

Sıçan Dalağında Makrofaj Alt Gruplarının Farklı Morfolojik Teknikler Kullanılarak İncelenmesi

Amaç: Dalak sahip olduğu özel morfololoji sayesinde, yaşlı hasarlı eritrositlerin yanı sıra patojenlerin kandan temizlenmesinde önemli rol üstlenir. Dalak makrofajları üzerinde yapılan çalışmalarda dala- ğın farklı bölgelerinde yerleşen makrofaj alt grupları tanımlanmıştır. Bu çalışmada sıçan dalağında bulunan makrofaj alt gruplarının intravenöz yolla verilen çini mürekkebine ve eritrositlere karşı fagositoz yeteneklerinin, lizozomal enzim aktivitelerinin ve ışık mikroskobu düzeyinde yapısal özelliklerinin değerlendirilmesi amaçlandı. Gereç ve Yöntem: Çalışmada 18 adet Wistar Albino tipi erişkin erkek sıçan kullanıldı. Hayvanlar üç eşit gruba ayrıldı. Birinci gruba herhangi bir işlem yapılmadı. ikinci ve üçüncü gruptaki hayvanlara kuyruk veninden ml/100 olacak şekilde çini mürekkebi verildi. Enjeksiyondan 30 dakika sonra ikinci gruptaki sıçanlar, 24 saat sonra üçüncü gruptakilerle birlikte birinci gruptaki sıçanlar sakrifiye edilerek dalakları çıkarıldı. Işık ve elektron mikroskobu için rutin tespit ve takip prosedürü uygulanarak elde edilen parafin ve araldit kesitlere çeşitli boyama teknikleri uygulandı. Lizozomal enzim aktivitelerinin belirlenmesi için enzim histokimya yöntemi kullanıldı. Bulgular: Sıçan dalağında marjinal bölgede yerleşen makrofajların karbon partiküllerine karşı zayıf fagositoz yeteneği gösterdiği, lizozomal enzim aktivitelerinin zayıf olduğu, morfolojik yönden kırmızı pulpada yerleşen makrofajlardan farklı oldukları belirlendi. Sonuç: Sıçan dalağının esas fagositoz yapan hücrelerinin kırmızı pulpa makrofajları olduğu sonucuna varıldı.

Investigation of Macrophage Subgroups in Rat Spleen by Using Different Morphological Techniques

Aim: As a result of its special morphology, the spleen plays an important role in cleaning of pathogens as well as old damaged erythrocytes from blood. Macrophage subgroups located in different areas of the spleen were identified through the studies performed on spleen macrophages. With this study we aimed to evaluate subgroups in rats spleen’s phagocytic and lysosomal enzyme activity for erythrocytes and intravenously administered India ink and macrophage subgroups structural properties assessed at light microscopy level. Material and Method: In this study, eighteen Wistar Albino adult male rats were used. Rats were divided into three equal groups. No application was made to the first group. India ink with a weight of 1 ml / 100 g were given to the second and third group animals from the tail vein. Thirty minutes after the injections, the animals of Group 2 were sacrificed. The animals of Group 3 were sacrificed twenty-four hours after the application simultaneously with Group 1 and their spleens were removed. Routine fixation and processing procedures applied for light and electron microscopy, and various staining techniques were performed to the paraffin and araldite sections. Enzyme histochemistry technique was used to determine lysosomal enzyme activities. Results: As a result, macrophages located in the marginal zone showed weak phagocytic activity to the carbon particles, their lysosomal enzyme activities were weak, and the macrophages located in red pulp were morphologically different. Conclusion: It was concluded that the main phagocytic cells of rat spleen are the red pulp macrophages

PDF

___

Mebius RE, Kraal G. Structure and 4. van Ewijk W, Nieuwenhuis P. Compart- function of the spleen. Nat Rev Immu- nol. 2005;5:606-616.
Schmidt EE1, MacDonald IC, Groom AC. Comparative aspects of splenic mic- rocirculatory pathways in mammals: the region bordering the white pulp. Scan- ning Microsc. 1993;7:613-628.
Kraal G. Cells in the marginal zone of the spleen. Int Rev Cytol. 1992;132:31-74. ments, domains and migration pathways of lymphoid cells in the splenic pulp. Experientia. 1985 15;41:199-208.
van Ewijk W, Nieuwenhuis P. Compartments, domains and migration pathways of lymphoid cells in the splenic pulp. Experientia. 1985 15;41:199-208.
Humphrey JH, Grennan D. Different macrophage populations distinguished by means of fluorescent polysaccharides. 7. den Haan JM, Kraal G. Innate immune Recognition and properties of marginal- zone macrophages. Eur J Immunol. 1981;11:221-228.
Prophet EB, Mills B, Arrington JB, Sobin LH, eds. Laboratory methods in histo- technology ( Armed Forces Institute of Pathology). 1st ed. Washington, D.C. : American Registry of Pathology; 1992:235-240. functions of macrophage subpopulations in the spleen. J Innate Immun. 2012; 4:437-445.
den Haan JM, Kraal G. Innate immune functions of macrophage subpopulations in the spleen. J Innate Immun. 2012; 4:437-445.
Borges da Silva H, Fonseca R, Pereira RM, et al. Splenic Macrophage Subsets and Their Function during Blood-Borne Infections. Front Immunol. 2015 22;6:480.
Elomaa O, Kangas M, Sahlberg C , et al. Cloning of a novel bacteria-binding re- ceptor structurally related to scavenger receptors and expressed in a subset of macrophages. Cell. 1995 24;80:603-609.
Chao D, MacPherson GG. Analysis of thymus-independent type 2 antigen upta- ke by marginal zone macrophages in thin slices of viable lymphoid tissue in vitro. Eur J Immunol. 1990;20:1451-1455.
Lee G. R , Bithell T.C, Foerster J et al., eds. Wintrobe's Clinical Hematology. 9th ed. Philadelphia: Lea & Febiger; 1992:311-322, 354.
Geijtenbeek TB1, Groot PC, Nolte MA, et al. Marginal zone macrophages express a murine homologue of DC-SIGN that captures blood-borne antigens in vivo. Blood. 2002 15;100:2908-2916.
Kraal G, Janse M. Marginal metallophilic cells of the mouse spleen identified by a monoclonal antibody. Immunology. 19. Junt T, Moseman EA, Iannacone M, et al. 1986;58:665-669.
Van Rooijen N, Kors N, vd Ende M, Dijkstra CD. Depletion and repopulation of macrophages in spleen and liver of rat after intravenous treatment with liposo- me-encapsulated dichloromethylene dip- hosphonate.. Cell Tissue Res. 1990;260:215-222.
Matsuno K, Fujii H, Kotani M. Splenic marginal-zone macrophages and marginal metallophils in rats and mice. Cell Tissue Res. 1986;246:263-269.
You Y, Myers RC, Freeberg L,et al. Marginal zone B cells regulate antigen capture by marginal zone macrophages. J Immunol. 2011 15;186:2172-2181.
Veerman AJ, van Ewijk W. White pulp compartments in the spleen of rats and mice. A light and electron microscopic study of lymphoid and non-lymphoid celltypes in T- and B-areas. Cell Tissue Res. 1975;156:417-441.
Phan TG, Green JA, Gray EE, et al. Immune complex relay by subcapsular si- nus macrophages and noncognate B cells drives antibody affinity maturation. Nat Immunol. 2009;10:786-793. Subcapsular sinus macrophages in lymph nodes clear lymph-borne viruses and pre- sent them to antiviral B cells. Nature. 2007 1;450:110-114.
Backer R, Schwandt T, Greuter M, et al. Effective collaboration between marginal metallophilic macrophages and CD8+ dendritic cells in the generation of cyto- toxic T cells. Proc Natl Acad Sci U S A. 2010 5;107:216-221.
Miyake Y, Asano K, Kaise H, et al. Criti- cal role of macrophages in the marginal zone in the suppression of immune res- ponses to apoptotic cell-associated anti- gens. J Clin Invest. 2007;117:2268-2278.
McGaha TL, Chen Y, Ravishankar B, et al. Marginal zone macrophages suppress innate and adaptive immunity to apopto- tic cells in the spleen. Blood. 2011 19;117:5403-5412.
King C, Tangye SG, Mackay CR. T folli- cular helper (TFH) cells in normal and dysregulated immune responses. Annu Rev Immunol. 2008;26:741-766.
Bratton DL, Henson PM. Apoptotic cell recognition: will the real phosphatidylse- rine receptor(s) please stand up? Curr Biol. 2008 22; 8:R76-79.