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Molecular cloning, characterization and tissue expression of CD4 in Chinese goose
Xiaoling Yan1, Fei Liu, Shun Chen
1Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, PR China.
Insights
This study identifies the CD4 gene in Chinese geese, revealing its structure and expression patterns in different tissues and during infection. Goose CD4 shows higher similarity to avian species, with varying expression levels critical for immune responses.
Area of Science:
- Immunology
- Genomics
- Veterinary Science
Background:
- CD4 protein is crucial for cell-mediated immunity, recognizing MHC class II molecules.
- Understanding CD4 in avian species like the Chinese goose (Anser cygnoides) is vital for comparative immunology.
Purpose of the Study:
- To clone and characterize the full-length CD4 cDNA in Chinese geese.
- To investigate the tissue-specific and developmental expression of goose CD4 mRNA.
- To analyze CD4 gene expression changes during Newcastle Disease Virus (NDV) infection.
Main Methods:
- Full-length cDNA cloning and sequencing of goose CD4.
- Bioinformatic analysis including multiple sequence alignment and phylogenetic analysis.
- Semi-quantitative RT-PCR for analyzing CD4 mRNA expression in various tissues and at different time points post-infection.
Main Results:
- The goose CD4 gene is 1940bp, encoding 480 amino acids with conserved functional domains.
- Phylogenetic analysis indicates goose CD4 is most similar to avian CD4 sequences.
- CD4 mRNA expression is highest in the thymus, with differential distribution in other tissues and age groups (gosling vs. adult).
- NGVEV infection significantly alters CD4 expression in the spleen, bursa of Fabricius, and Harderian gland, with dynamic changes in the small intestine.
Conclusions:
- The study provides the first detailed characterization of goose CD4, highlighting its evolutionary conservation within avian species.
- Tissue-specific expression patterns suggest a role for CD4 in both primary and secondary lymphoid organs.
- CD4 gene expression is modulated by viral infection, indicating its involvement in the goose's immune response to pathogens.
Abstract:
CD4 protein is a single chain transmembrane glycoprotein belonging to the immunoglobulin superfamily that recognizes the major histocompatibility complex (MHC) class II molecules. It plays an important role in cell-mediated immunity. Here, the full-length cDNA of CD4 in Chinese goose (Anser cygnoides) was cloned and identified. The goose CD4 is 1940bp in length and encodes a single open-reading frame of 480 amino acids. The putative amino acid sequence of goose CD4 consisted of a signal peptide, four potential N-glycosylation sites, a transmembrane region and a cytoplasmic tail. The multiple sequence alignment and phylogenetic analysis suggested that goose CD4 shared a higher similarity with avian than other vertebrates. Semi-quantitative RT-PCR analysis showed that the highest level of CD4 mRNA transcripts was presented in the thymus, and relatively lower in the spleen, small intestine, brain and trachea. Low expression was seen in the bursa of fabricius, cecal tonsil, cecum, skin, lung, kidney and liver. In gosling, the CD4 transcript was expressed with high abundance in the thymus, and relatively lower in the spleen, cecal tonsil and small intestine. However, in adult goose, high expression was seen in the thymus, spleen and cecum, and relatively lower in the cecal tonsil and small intestine. During the course of NGVEV infection, the obvious increase in CD4 gene expression was observed in the spleen, bursa of fabricius and harderian gland. Interestingly, a notable decrease in CD4 mRNA expression in the small intestine at 5d PI and followed by an increase of that at 19d PI were shown.
