Immunohistochemical Examination of TNF-α and SAA Expressions in Goats Infected with Mycobacterium avium subspecies paratuberculosis

Öz

Paratuberculosis (Johne Disease), caused by Mycobacterium avium subspecies paratuberculosis (MAP) and commonly seen in ruminants, is a contagious disease that causes chronic diarrhea and serious economic losses. In this study, the presence of pro-inflammatory cytokine and acute phase protein expressions in lesional tissues of naturally infected goats with MAP was investigated. For this purpose, paratuberculosis suspected goats complaining of chronic diarrhea and excessive weight loss were examined by ELISA method for Mycobacterium avium subspecies paratuberculosis infection. After sacrificing 20 animals found to be infected with MAP, tissue samples taken from the intestines and lymph nodes were stained with Hematoxylin-Eosin (HE) for histopathological examination and Ziehl-Nelsen (ZN) to show acid-fast mycobacterium. Immunohistochemical staining was performed to examine the expressions of TNF-α as a pro-inflammatory cytokine and SAA as an acute phase protein. Macroscopic examination revealed in serous fat atrophy, mesenteric lymphadenitis, and granulomatous enteritis in the intestines. Histopathological examination revealed atrophy in villi, degeneration of enterocytes, inflammatory cell infiltration consisting mostly of mononuclear leukocytes in the mucosa, and epithelioid macrophage aggregation. Lymphoid hyperplasia and epithelioid histocyst aggregates were found in the mesenterial lymph nodes. In staining with ZN revealed the presence of bright red acid-fast agents in the intestine and mesenteric lymph nodes. In immunohistochemical examination, TNF-α and SAA expressions were observed in all samples. With this study, for the first time, the expressions of TNF-α, a proinflammatory cytokine, and SAA, an acute phase protein, were shown in the lesional tissues of MAP-infected animals, contributing to the immunopathogenesis of the disease.

Anahtar Kelimeler

• Goat
• Mycobacterium avium subspecies paratuberculosis
• paratuberculosis
• SAA
• TNF-α

Mycobacterium avium subspecies paratuberculosis ile Enfekte Keçilerde TNF-α ve SAA Ekpresyonlarının İmmünohistokimyasal İncelenmesi

Öz

Etkeni Mycobacterium avium subspecies paratuberculosis (MAP) olan ve geviş getirenlerde yaygın olarak görülen paratüberküloz (Johne Disease) kronik ishal ile ciddi ekonomik kayba neden olan bulaşıcı bir hastalıktır. Bu çalışmada MAP ile doğal enfekte keçilerin lezyonlu dokularında pro-inflmatuvar sitokin ve akut faz protein ekpresyonlarının varlığı araştırıldı. Bunun için kronik ishal ve aşırı zayıflama şikayeti bulunan paratüberküloz şüpheli keçiler Mycobacterium avium subspecies paratuberculosis enfeksiyonu yönünden ELISA yöntemiyle incelendi. MAP ile enfekte olduğu saptanan 20 hayvanın kesime sevki sonrası sakrifiye edilmeleri ile bağırsak ve lenf yumrularından alınan doku örnekleri histopatolojik inceleme için Hematoksilen-Eozin (HE) ve asit-fast mycobacteriumları göstermek için Ziehl-Nelsen (ZN) ile boyandı. Pro-inflamatuvar sitokin olarak Tümör Nekroz Faktör (TNF-α) ve akut faz protein olarak Serum Amiloid A (SAA) ekpresyonlarını incelemek için immünohistokimyasal boyama yapıldı. Makroskobik incelemede seröz yağ atrofisi, mezenteriyal lenfadenit ve bağırsaklarda granülamatoz enteritis görüldü. Histopatolojik incelemede villuslarda atrofi, enterositlerde dejenerasyon, mukozada çoğunluğu mononükleer lökositlerden oluşan yangı hücre infiltrasyonu ve epiteloid makrofaj agregasyonu gözlendi. Mezenteriyal lenf düğümlerinde lenfoid hiperplazi ve epiteloid histoyosit yığınlarına rastlandı. ZN boyamada bağırsak ve mezenterik lenf düğümlerinde parlak kırmızı renkte asit-fast etkenlerin varlığı saptandı. İmmünohistokimyasal incelemede TNF-α ve SAA ekpresyonlarının tüm örneklerde meydana geldiği gözlendi. Bu çalışma ile ilk defa MAP ile enfekte hayvanların lezyonlu dokularında bir proinflamatuar sitokin olan TNF-α ve bir akut faz proteini olan SAA ekpresyonları gösterilerek hastalığın immünpatogenezine katkı sağlandı.

Anahtar Kelimeler

• Keçi
• Mycobacterium avium subsp. paratuberculosis
• paratüberküloz
• SAA
• TNF-α

Kaynakça

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