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    Comparison of immune responses elicited by adjuvanted tachyzoite lysate vaccines developed from two different toxoplasma gondii strains isolated in Turkey [Türkiye'de ïzole edilen i·ki farkli toxoplasma gondii suşundan üretilen adjuvanli takizoit eriyik protein aşilarinin uyardigi immün yanitlarin karşilaş tirilmasi]
    (Ankara Microbiology Society, 2013) Polat C.; Sultan Gülçe I.Z.; Döşkaya M.; Hüseyin C.A.N.; Caner A.; Degirmenci A.; Balcan E.; Gürüz Y.
    Toxoplasma gondii the causative agent of toxoplasmosis is an obligate intracellular parasite with a wide host range including all warm-blooded animals and birds. T.gondii infection causes congenital toxoplasmosis in newborns and this may lead to fetal anomalies, retinochoroiditis leading to blindness, lethal toxoplasmic encephalitis in immune compromised patients, and organ failure in transplantation patients. The pathogenesis of toxoplasmosis change due to differences in the specific immune response elicited by diverse T.gondii strains. The protective immunity against toxoplasmosis is conferred by cellular immune responses. In the present study, two different strains isolated from Turkey named T.gondii Ankara and Ege were used to evaluate the types of humoral and cellular immune responses elicited by adjuvanted tachyzoite protein vaccines in an animal model. In the study, 6-8 weeks old female BALB/c mice were used and six study groups (each contains three mice) were composed for vaccination. The first and second groups were vaccinated with T.gondii Ankara and Ege (TAnkPE and TEgePE, respectively) tacyhzoite lysates, the third and fourth groups were vaccinated by tacyhzoite lysates adjuvanted with Freund's adjuvant (TAnkPE-Freund; TEgePE-Freund, respectively). The fifth and sixth groups were vaccinated with PBS and Freund's adjuvant as controls. Immunization of the animals was performed two times at three weeks intervals. The serum samples were collected before vaccination and after each vaccination to determine the IgG response by Western blotting, and IgG1 and IgG2a responses by ELISA. To determine the cellular immune response, CD8/CD4 cell ratio, intracellular IFN-y and IL-4 levels were determined in stimulated spleen cells grown in cell culture systems by flow cytometry. Toxoplasma IgG antibodies were only detected in TAnkPE-Freund group. IgG1 and IgG2a responses did not increase in any vaccination groups and there was not any polarization towards IgG1 or IgG2a. There was no significant increase in CD8/CD4 ratio of stimulated spleen cells. IFN-y level was increased in only TAnkPE-Freund vaccination group, however IL-4 levels were increased in TAnkPE-Freund, TEgePE-Freund and TEgePE groups. Our data showed that TAnkPE-Freund vaccine led to increase in IgG and IFN-y responses in BALB/c mice, however, tachyzoite lysate vaccines developed in this study did not induce sufficient protective immune response against toxoplasmosis. Thus, use of specific immunogenic proteins must be taken into consideration in the future vaccine development studies against toxoplasmosis.
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    Ultrastructural dynamics of transendothelial migration of lymphocytes through high endothelial venules (HEVs) of the mucosa associated peyer’s patches
    (Croatian Society of Natural Sciences, 2017) Balcan E.; Karaçali S.
    Background and Purpose: Although the pivotal role of high endothelial venules (HEVs) in the migration of leukocytes from the blood into the secondary lymphoid parenchyma is well established, conflicting ideas concerning the cellular dynamics both of leukocytes and endothelial cells throughout the migratory processes have been present. Here we focused specifically on the cellular dynamics of HEVs from Peyer’s patches in an ultrastructural perspective. Materials and Methods: In order to determine the microstructural organization of transendothelial migration we used conventional methods for transmission electron microscopy. Results: Our results indicate that both lymphocytes and endothelial cells are highly active in the processes of transmigration steps, and a series of morphological and cellular alterations can occur depending upon their activity. Various types of cellular protrusions provide a direct contact between luminal lymphocytes and the endothelial cells at the initial phases of the migration. The endothelial protrusions subsequently embrace the lymphocytes and guide them into lymphoid stroma during the transcellular migration. Meanwhile, different sizes of vesicles show different cellular localizations according to their roles. The vesicles which are clustered near the lateral borders and the stand alone ones found only in the abluminal surfaces of endothelial cells might be involved in the paracellular migration. Concurrently, the other types of vesicles were smaller and appeared in the lateral border of the endothelial cells. Differ from the clustered and abluminally localized vesicles, they were closely related with plasma membranes. Conclusions: These results indicated that not only adhesion molecules, but also cellular dynamics of leukocytes and endothelial cells regulate the leukocyte traffic into lymphoid stroma or vice versa. © 2017, Kanagawa Prefectural Museum of Natural History. All rights reserved.