Bruseloz'da spesifik T hücre yanıtının regülasyonu
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Date
2006
Authors
Kazak, Esra
Journal Title
Journal ISSN
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Publisher
Uludağ Üniversitesi
Abstract
Brucella, insanlarda endokardit, artrit, osteomiyelit, menenjite yol açan gram negatif hücre içi bakteridir. Hastalığın kronikleşme riski ve yüksek morbiditesi nedeni ile etkili aşı ve yeni tanı reaktanlarının geliştirilmesine ihtiyaç duyulmuştur. Bu amaçla Brucella antijenlerinden yeni sentetik peptitler geliştirilmiş ve hayvan modellerinde denenmiştir. Ancak bu peptidlere karşı insan immün sisteminin yanıtına ait yeterli çalışma yoktur. Çalışmamızda B. abortus’dan elde edilen L7/L12 ve GAPDH peptitlerine karşı Bruselloz’lu hastaların periferal kan lenfositlerinin profilleri ve sitokin yanıtının araştırılması amaçladık. Akut Bruselloz’lu ve sağlıklı hastaların kan lenfositlerinde bu antijenlere yanıt olarak ortaya çıkan lenfosit alt gruplarındaki değişiklikleri akım sitometrisi ile ve hücre dışı sitokin üretimini bu hücrelerin kültür süpernatantlarında ELISA yöntemi ile araştırdık. Çalışmaya 25 Akut Bruselloz hastası ve 15 sağlıklı birey alındı. Bu kişilerden elde edilen periferal mononükleer hücreler PHA, L7/L12, GAPDH ile uyarıldıktan sonra lenfosit alt grupları ve hücre dışı sitokin (IFN-γ, IL-4, IL10) miktarı ölçüldü. Uyarı sonucu Bruselloz’lu hastalarda hücre dışı IFN-γ seviyeleri yüksek tespit edildi. Gruplar arasında uyarı sonrası IL-4, IL-10 seviyeleri arasında farklılık yoktu. Çalışmada ayrıca CD3+/CD4+, CD3+/CD8+, CD4+/CD25+, CD4+/CD27-, CD3+/CD152+(CTLA4), CD3+/CD69+ lenfosit dağılımı araştırıldı. Bu peptidler ile uyarı sonucunda hastalarda CD3+/CD69+ lenfosit oranının arttığı saptandı. Ancak diğer lenfosit alt gruplarının dağılımında hasta ve sağlıklı gruplar arasında farklılık tespit edilmedi. Bu çalışmalarda elde edilen veriler ışığında Bruselloz’da koruyucu immüniteden Th1 tip immünitenin sorumlu olduğunu, GAPDH ve/veya L7/L12’nin yeni Bruselloz aşı preparasyonlarında; bu peptitler ile uyarıyı takiben IFN-γ yapımını temel alan spesifik hücresel immün yanıtı ölçen immünolojik tanı testlerinde kullanılabileceğini söyleyebiliriz. Bununla birlikte GAPDH ve L7/L12’nin kompleks immünomodülatör etkilerini açıklığa kavuşturacak yeni ve daha ileri çalışmalara gereksinim vardır.
Brucella is a gram-negative intracellular bacterium that leads to endocarditis, arthritis, meningitidis and osteomyelitis in humans. Because of the chronicity risk of the disease and high morbidity, there are requirements for effective vaccines and spesific diagnostic reagents to eradicate brucellosis. For that purpose novel synthetic peptides derived from Brucella antigens were generated and tested in animal models. However there is a little known about the human immune response to these peptides. In this study we investigated the profiles of the peripheral blood lymphocytes and cytokine responses of Acute Brucellosis patients and healthy subjects in response to two novel peptides derived from Brucella; GAPDH and L7/L12 by evaluating changes in lymphocyte subpopulations and by measuring the amount of cytokine productions in culture supernatants using flow cytometry and ELISA, respectively. Twenty-five Acute Brucellosis patients and fifteen healthy subjects were included in the study. The peripheral blood mononuclear cells obtained from Acute Brucellosis patients (study group) and healthy subjects (control group) were stimulated with PHA, L7/L12, GAPDH prior to the measurement of extracellular cytokines (IFN-γ, IL-4, IL-10) and evaluation of the lymphocytes subgroups. As a result of stimulation, extracellular IFN-γ levels increased in brucellosis patients. There were no differences in the levels of IL-4, IL-10 under the stimulation with these peptides between the groups. Also CD3+/CD4+, CD3+/CD8+, CD4+/CD25+, CD4+/CD27-, CD3+/CD152+(CTLA4), CD3+/CD69+ lymphoctes were counted. The CD3+/CD69+ counts in patients increased under the stimulation with L7/L12, GAPDH. However there were no statistically significant differences in the other lymphocytes subpopulations between the healthy and patient groups. In the light of the data obtained in our study, it can be suggested that Th1 type spesific cellular immunity is responsible for protective immunity in Brucellosis. The more potent vaccines may be developed by including L7/L12, GAPDH and diagnostic tests using these peptides may be evaluated. However, there are needs for new and further studies which reveals the complex immunomodulatory effects of GAPDH and L7/L12.
Brucella is a gram-negative intracellular bacterium that leads to endocarditis, arthritis, meningitidis and osteomyelitis in humans. Because of the chronicity risk of the disease and high morbidity, there are requirements for effective vaccines and spesific diagnostic reagents to eradicate brucellosis. For that purpose novel synthetic peptides derived from Brucella antigens were generated and tested in animal models. However there is a little known about the human immune response to these peptides. In this study we investigated the profiles of the peripheral blood lymphocytes and cytokine responses of Acute Brucellosis patients and healthy subjects in response to two novel peptides derived from Brucella; GAPDH and L7/L12 by evaluating changes in lymphocyte subpopulations and by measuring the amount of cytokine productions in culture supernatants using flow cytometry and ELISA, respectively. Twenty-five Acute Brucellosis patients and fifteen healthy subjects were included in the study. The peripheral blood mononuclear cells obtained from Acute Brucellosis patients (study group) and healthy subjects (control group) were stimulated with PHA, L7/L12, GAPDH prior to the measurement of extracellular cytokines (IFN-γ, IL-4, IL-10) and evaluation of the lymphocytes subgroups. As a result of stimulation, extracellular IFN-γ levels increased in brucellosis patients. There were no differences in the levels of IL-4, IL-10 under the stimulation with these peptides between the groups. Also CD3+/CD4+, CD3+/CD8+, CD4+/CD25+, CD4+/CD27-, CD3+/CD152+(CTLA4), CD3+/CD69+ lymphoctes were counted. The CD3+/CD69+ counts in patients increased under the stimulation with L7/L12, GAPDH. However there were no statistically significant differences in the other lymphocytes subpopulations between the healthy and patient groups. In the light of the data obtained in our study, it can be suggested that Th1 type spesific cellular immunity is responsible for protective immunity in Brucellosis. The more potent vaccines may be developed by including L7/L12, GAPDH and diagnostic tests using these peptides may be evaluated. However, there are needs for new and further studies which reveals the complex immunomodulatory effects of GAPDH and L7/L12.
Description
Keywords
Akut Bruselloz, GAPDH, L7/L12, Sitokin, Lenfosit Alt Grupları, IFN-γ, IL-4, IL-10, Acute Brucellosis, Cytokine, Lymphocyte Subpopulation
Citation
Kazak, E. (2006). Bruseloz'da spesifik T hücre yanıtının regülasyonu. Yayınlanmamış uzmanlık tezi. Uludağ Üniversitesi Tıp Fakültesi.