Studying the Cell Specificity and Differential Immune Activation by Chicken Type III Interferons (IFN-λ3)

Malakar, Dipendu (2025) Studying the Cell Specificity and Differential Immune Activation by Chicken Type III Interferons (IFN-λ3). Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS Dissertation of Dipendu Malakar (20MS105)) 20MS105_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (2MB)

Abstract

The poultry industry has always been subjected to threats from viral infection, especially avian influenza viruses, which are notoriously hard to control with their high mutation rates and low efficacy of traditional vaccination methods. As an alternative to control influenza infections, the use of antiviral cytokines like interferons have been recognized. Among them, recent studies in mammalian models have described type III interferon, especially IFN-λ3, to induce a robust antiviral activity with minimal inflammation through the virtue of its epithelial cell-specific receptor expression. Unlike various mammalian models, the function of type III IFNs in avian immune system is not well documented. This study intends to discover whether the conclusions about Type III IFN’s epithelial cell-specific behavior holds true for avian models, using In Vitro experiments on different chicken primary cells. We externally produced recombinant chicken IFN-λ3(rChIFN-λ3) proteins in E. coli expression system to treat primary chicken embryonic intestinal cells (CEICs) and chicken splenocytes with varying concentrations. After that we examined the expression of four important interferon-stimulated genes (ISGs)- Viperin, OASL, Mx1, IFITM3 and a type III IFN receptor subunit IL-28RA, using RT-PCR to evaluate any difference in immune response generated by the rChIFN-λ3. The results indicated a noticeable baseline (without any rChIFN-λ3 treatment) expression of IL-28RA in CEICs than in splenocytes, supporting our initial hypothesis. Furthermore, a significant increase in downstream ISGs with increasing dose of rChIFN-λ3 was observed in CEICs but not in case of chicken splenocytes. This indicated the epithelial cell specific activity type III IFN (chIFN-λ3), which is in line with the mammalian systems.

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