Abstract
BackgroundCirrhosis of liver is a pathological condition, wherein functions of liver are impaired by chronic liver exploitations. Due to decrease in synthetic capacity, expressions of plasma proteins tend to decrease in blood stream. Serpins (Serine protease inhibitors) are class of plasma proteins expressed from liver with structural similarities and diverse functions. SERPINA4 (Kallistatin) is a multifunctional serpin clade A protein expressed from liver and concentration in serum is the reflection of extent of liver dysfunction. ObjectiveTo identify interference of other serpins by immunological cross reactivity with SERPINA4 in cirrhotic liver and healthy subjects. Materials and methodsBlood samples were collected from 20 subjects (10 cirrhotic liver, 10 healthy) from R.L. Jalappa Hospital and Research Centre, Kolar, Karnataka, India. Separation of proteins was carried out by SDS-PAGE. Cross reactivity study was analyzed using western blot. ResultsProteins present in cirrhotic liver and healthy subject's serum were separated by SDS PAGE. There was no band detection on both (cirrhotic liver and healthy) PVDF (polyvinylidene diflouride) membranes. However, a significant band was observed with recombinant kallistatin. ConclusionStructurally similar serpins with minor amino acid sequence similarities did not show any immunological cross reactivity with SERPINA4 due to non identical epitope in cirrhotic liver and healthy subjects. Present study revealed that there is no interference of serpins for immunological reactions in quantitative estimation of kallistatin which needs further validation.
Highlights
Cirrhosis of liver is a pathological condition characterized by diffuse fibrosis, severe disruption of intra hepatic arterial and venous flow, portal hypertension and liver failure resulting from varied etiologies of chronic liver diseases [1]
Recombinant kallistatin was spotted on another SDS-PAGE with pooled and concentrated samples of cirrhotic liver and healthy subjects
Western blot analysis allowed identification of cross reactivity of serpins in diseased and healthy samples using monoclonal antibodies specific for kallistatin followed by secondary antibodies conjugated with HRP
Summary
Cirrhosis of liver is a pathological condition characterized by diffuse fibrosis, severe disruption of intra hepatic arterial and venous flow, portal hypertension and liver failure resulting from varied etiologies of chronic liver diseases [1]. Serpins (Serine Protease Inhibitors) are class of plasma proteins that have similar structure and diverse functions. Majority of clade A serpins are localized on chromosome 14 which are expressed from liver. Serpins (Serine protease inhibitors) are class of plasma proteins expressed from liver with structural similarities and diverse functions. SERPINA4 (Kallistatin) is a multifunctional serpin clade A protein expressed from liver and concentration in serum is the reflection of extent of liver dysfunction. Objective: To identify interference of other serpins by immunological cross reactivity with SERPINA4 in cirrhotic liver and healthy subjects. Results: Proteins present in cirrhotic liver and healthy subject's serum were separated by SDS PAGE. Conclusion: Structurally similar serpins with minor amino acid sequence similarities did not show any immunological cross reactivity with SERPINA4 due to non identical epitope in cirrhotic liver and healthy subjects. Present study revealed that there is no interference of serpins for immunological reactions in quantitative estimation of kallistatin which needs further validation
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