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Published on: November 5, 2012
Molecular cloning and expression analysis of pig CD79alpha
Suk Jun Lee1, Sang Joon Kim, Chung-Gyu Park
1Division of Biotechnology, School of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea.
Insights
Researchers cloned pig CD79a cDNA, revealing conserved signaling motifs crucial for B cell receptor function. Pig CD79alpha is primarily expressed in lymphoid tissues, aiding in B cell signaling research.
Area of Science:
- Immunology
- Molecular Biology
- Veterinary Science
Background:
- The CD79alpha (immunoglobulin alpha, Igalpha) protein is a key component of the B cell receptor (BCR) complex.
- CD79alpha heterodimerizes with CD79beta (Igbeta) to mediate B cell signaling pathways.
- Understanding CD79alpha in pigs is vital for comparative immunology and disease research.
Purpose of the Study:
- To clone and characterize the pig CD79a cDNA sequence.
- To investigate the tissue distribution and expression patterns of pig CD79a mRNA.
- To confirm the utility of a human CD79alpha antibody for detecting pig CD79alpha.
Main Methods:
- Reverse transcription-polymerase chain reaction (RT-PCR) for cDNA cloning and mRNA detection.
- Bioinformatic analysis of the deduced amino acid sequence and comparison with mammalian orthologs.
- Flow cytometry and immunohistochemistry using a human CD79alpha antibody on pig tissues.
Main Results:
- The complete cDNA sequence of pig Cd79a was determined, encoding 223 amino acids with significant homology to mammalian CD79alpha.
- Conserved transmembrane and cytoplasmic domains, including the immunoreceptor tyrosine-based activation motif (ITAM), were identified.
- Pig Cd79a mRNA expression was highest in the mesenteric lymph node and spleen, with lower levels in other lymphoid and non-lymphoid tissues.
- A human CD79alpha antibody successfully detected pig CD79alpha on B cells and in spleen tissue, localized to the follicular mantle zone.
Conclusions:
- The cloned pig CD79a sequence provides a foundation for further immunological studies in pigs.
- The conserved structure suggests similar functional roles in B cell signaling across mammalian species.
- The expression patterns highlight the importance of lymphoid organs in pig CD79alpha function.
- The validated antibody offers a tool for future research into pig infectious diseases and neoplasms involving B cells.
Abstract:
The CD79alpha (immunoglobulin alpha, Igalpha), a part of B cell receptor (BCR) complex, forms a heterodimer with CD79beta (Igbeta) and plays an important role in the B cell signaling. In this study, we have cloned pig Cd79a cDNA using RT-PCR and determined the complete cDNA sequence of pig Cd79a. Pig Cd79a cDNA contains an open reading frame (672bp) encoding 223 amino acids. The putative amino acid identity of pig CD79alpha with those of human, cattle and mouse are 70.4, 81.4, and 67.7%, respectively. Alignment of the CD79alpha amino acid sequence with those of mammalian species showed that the extracellular domain is the most divergent, whereas transmembrane region and cytoplasmic tail including immunoreceptor tyrosine-based activation motif (ITAM) are largely conserved. Pig Cd79a mRNA was detected mainly in lymphoid tissues by RT-PCR. The highest level of Cd79a mRNA expression was observed in mesenteric lymph node and spleen. Relatively low level of Cd79a mRNA expression was observed in lung, thymus and small intestine. The lowest level of Cd79a mRNA expression was observed in large intestine. Flow cytometry analyses demonstrated that human CD79alpha antibody recognizes a CD79alpha in pig B cells. Further, immunohistochemistry analysis using human CD79alpha antibody on pig spleen was revealed that CD79alpha is strongly expressed in the follicular mantle zone rather than in the germinal center. Future study will be focused on defining the functional role of CD79alpha during the course of pig infectious diseases and the formation of neoplasm.
