BCL10 as a new candidate gene for immune response in pigs: cloning, expression and association analysis J. Huang,* G.-J. Ma,* N.-N. Sun,† Z.-F. Wu,† X.-Y. Li* & S.-H. Zhao* Summary BCL10 is an apoptotic regulatory molecule identified through its direct involvement in t(1; 14)(p22; q32) of mucosa-associated lymphoid tissue lymphoma, and was implicated in the pathogenesis of this and several other tumour types. BCL10 was recognized as an antigen receptor-specific regulator of NF-jB, which showed close association with immune responses. In this study, we cloned and characterized BCL10 from the porcine spleen and analysed its genomic structure. BCL10 was mapped to SSC4q21–q23 by the IMpRH panels, it is closely linked to the marker S0161 and SW1461. This gene has three exons and two introns. Reverse trans- criptase-polymerase chain reaction analyses showed that BCL10 was widely expressed in all the examined tissues. Transient transfection indicated that porcine BCL10 was located in cytoplasm in Pig Kidney Epithe- lial cells. BCL10 gene displays the opposite expression trend between the two treatments mimic virus and bacteria of polyriboinosinic-polyribocytidylic acid (Poly I:C) and lipopolysaccharide (LPS). The level of the BCL10 mRNA was up-regulated during 12–24 h and peaking at 48 h when treated with LPS, whereas it was down-regulated during 0–48 h and highest at 0 h (cells without treating with Poly I:C) when treated with Poly I:C. One single nucleotide polymorphism (SNP) site was identified in the 3¢-untranslated region of porcine BCL10. Association analysis revealed that this SNP was significantly associated with intermediate cell mass (eosinophile granulocyte, basophile granulocyte and histoleucocyte) percentage, absolute intermediate cell mass count and mean red blood cell volume of 0-day- old pigs, and red blood cell count of 17-day-old pigs (P < 0.05), and also had significant associations with red blood cell count and haemoglobin concentration of 32-day-old pigs (P < 0.01). Introduction The BCL10 gene was originally identified through its recurrent involvement in t(1;14)(p22;q32) of the mucosa-associated lymphoid tissue (MALT) lym- phoma. It was translocated to the immunoglobulin heavy chain locus in MALT (Willis et al., 1999). Human BCL10 gene encodes a protein of 233 amino acids with residues 13–101 forming a caspase recruit- ment domain (CARD), which was found in a number of apoptotic regulatory molecules (Hofmann et al., 1997). The N-terminal CARD domain is required for its interaction with the hematopoietic-specific mem- brane-associated guanylate kinase (MAGUK) protein Carma1 while the C-terminal Ser ⁄ Thr rich region (res- idues 97–233) is phosphorylated (Bertin et al., 2001; Gaide et al., 2001). Carma1 is lipid raft-associated, co-localizes with the T-cell receptor (TCR) in T cells and is required for TCR-activation of NF-jB (Egawa et al., 2003). Bcl10 adaptor protein was recognized as an antigen receptor-specific regulator of NF-jB. Also some results showed that Truncated BCL10 lost the pro-apoptotic activity but retained the ability of NF- jB activation and, moreover, gained functional enhancement of malignant transformation (Willis et al., 1999). Bcl10 was associated with NF-jB activa- tion in cell surface-proximal signalling aspects, and other studies have suggested the protein acts upstream of IjB (Bouchier-Hayes et al., 2001; Cheng et al., 2002). The interaction of Bcl10 with several signalling partners in the regulation of NF-jB activation indi- cates that it may be essential in multiple physiological functions. Using knockout mice, several researchers found that Bcl10 is essential for the development of all mature B- cell subsets, including follicular, marginal zone and B1 B cells (Xue et al., 2003), and also associated with the * Key Lab of Agricultural Animal Genetics, Breeding, and Reproduc- tion of Ministry of Education & Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, China and † College of Animal Science and Technology, Huanan Agricultural University, Guangzhou 510642, China Received 3 January 2009; revised 2 September 2009; accepted 21 December 2009 Correspondence: Shu-Hong Zhao, Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China. Tel.: +86 27 87284161; Fax: +86 27 87280408; E-mail: shzhao@mail.hzau.edu.cn; Zhen-Fang Wu, College of Animal Science and Technology, Huanan Agricultural University, Guangzhou 510642, China. Tel.: 86-766-2986001; Fax: +86 766 2986008; E-mail: wzfemail@163.com ª 2010 Blackwell Publishing Ltd, International Journal of Immunogenetics 37, 103–110 103 doi: 10.1111/j.1744-313X.2010.00898.x