Gene Expression Analysis of Swine Leukocyte Antigen (SLA-1 and SLA-2) Involved in Porcine Pre-Weaning and Post-Weaning Diarrhea in Nueva Ecija, Philippines
Mary Rose D. Uy1§, Gemerlyn G. Garcia2§, Jeffrey P. Aquino2,
Joan F. Sampang2, Reginaldo V. Abuyuan2, and Claro N. Mingala1,3§*
1Philippine Carabao Center National Headquarters and Gene Pool,
Science City of Muñoz 3120, Nueva Ecija, Philippines
2College of Veterinary Science and Medicine, Central Luzon State University,
Science City of Muñoz 3120, Nueva Ecija, Philippines
3Department of Animal Science, College of Agriculture,
Central Luzon State University, Science City of Muñoz 3120, Nueva Ecija, Philippines
§These authors contributed equally to this work
The immune responses of two breeds of piglets to diarrhea at pre-weaning and post-weaning were evaluated in terms of the relative quantification of Major Histocompatibility Complex (MHC) glycoproteins represented by the swine leukocyte antigen (SLA) class I. The expression of SLA-1 and SLA-2 genes of diarrheic and non-diarrheic Native and Large White piglets were measured using real time polymerase chain reaction (qPCR). Blood samples from 20 Native and 20 Large White piglets were used in this study. It is comprised of 5 Native piglets with clinical signs of diarrhea and 5 Native piglets with no diarrhea at pre-weaning. Same number of piglets were used for Native piglets at post-weaning and Large White piglets at pre-and post-weaning periods. The cDNA samples were amplified using primers for SLA-1 and SLA-1 alleles having amplicon sizes of 217 bp and 126 bp, respectively. Factors that were considered in the study include breed and status of piglets. Relative quantification was done using comparative threshold cycle (CT) method. Significantly higher levels of SLA-1 were noted in diarrheic pigs compared to those of non-diarrheic piglets (P=0.040) of the Native and Large White breeds at pre-weaning period. This observation was not analogous with the non-significant differences in SLA-2 expression, deduced as SLA-linked immune responses of piglets from the Native and Large White breeds with and without diarrhea observed at pre-weaning and post-weaning stages. The upregulation of SLA-1 in piglets with diarrhea at pre-weaning in the two breeds of swine examined the potential role of SLA-1 in the host’s response to diarrhea. These data associate the significance of the SLA-1 gene as a marker for diarrhea in pre-weaning piglets.
Porcine pre- and post-weaning diarrhea poses a serious threat to the rapid development of the pig industry. It leads to reduced weight gain, increased mortality, and morbidity in piglets (Jonach et al. 2014).
The gastrointestinal tract hosts a multitude of microorganisms. Bacteria can interact in a competitive manner and any disturbance in the microbiota can cause bacterial proliferation and disease (Hao 2004). A number of microorganisms and parasites have been identified to be associated with diarrhea in piglets and those include enterotoxigenic Escherichia coli (ETEC), Clostridium perfringens type C, Cryptosporidium spp., Giardia spp., Cystoisospora suis, and Strongyloides ransomi (Maddox-Hyttel et al. 2006; Brown et al. 2007). Pre-weaned and post-weaned piglets are vulnerable to infectious enteric diseases due to immunodeficiency and incomplete gut microbiota at birth (Bauer et al. 2006). Morbidity and mortality in neonatal piglets due to pre- and post-weaning reportedly . . . . . read more
ANDO A, CHARDON P. 2006. Gene organization and polymorphism of the swine major histocompatibility complex. Anim Sci J 77: 127-137.
ANDO A, KAWATA H, SHIGENARI A, ANZAI T, OTA M, KATSUYAMA Y, SADA M, GOTO R, TAKESHIMA S, AIDA Y, IWANAGA T, FUJIMURA N, SUZUKI Y, GOJOBORI T, INOKO H. 2004. Genetic polymorphism of the swine major histocompatibility complex (SLA) class I genes, SLA-1, -2 and -3. Immunogenetics 55: 583-593.
BAO WB, YE L, PAN ZY, ZHU J, DU ZD, ZHU GQ, HUANG XG, WU SL. 2012. Microarray analysis of differential gene expression in sensitive and resistant pig to Escherichia coli F18. Anim Genet 43: 525-534.
BAUER E, WILLIAMS BA, SMIDT H, VERSTEGEN MWA, MOSENTHIN R. 2006. Influence of the gastrointestinal microbiota on development of the immune system in young animals. Curr Issues Intest Microbiol 7: 35-52.
BROWN CC, BAKER DC, BARKER IK. 2007. Infectious and parasitic diseases of the gastrointestinal tract. In: Pathology of Domestic Animals 2, 5th ed. Grant Maxie M (ed). Philadelphia: Saunders/Elsevier. p.170-177.
CHARDON P, ROGEL-GAILLARD C, CATTOLICO L, DUPRAT S, VAIMAN M, RENARD C. 2001. Sequence of the swine major histocompatibility complex region containing all non-classical class I genes. Tissue Antigens 57(1): 55-65.
[CDC-OID] Centers for Disease Control and Prevention - Office of Infectious Disease. 2013. Antibiotic resistance threats in the United States. Retrieved from http://www.cdc.gov/drugresistance/threat-report-2013 on 28 Jan 2018.
DALE D, FEDERMAN D. 1998. Scientific American Medicine. Encyclopedia of Life Sciences: Macmillan Publishers Ltd., Nature Publishing Group.
FAIRBROTHER JM, NADEAU E, GYLES CL. 2005. Escherichia coli in post-weaning diarrhea in pigs: an update on bacterial types, pathogenesis, and prevention strategies. Anim Health Res Rev 6(1):17-39.
FRELS WI, BORDALLO C, GOLDING H, ROSENBERG A, RUDIKOFF S, SINGER S. 1990. Expression of a class I MHC transgene: regulation by a tissue specific negative regulation DNA sequence element. New Biol 67: 211-218.
HAO W-L. 2004. Microflora of the gastrointestinal tract: A review. In: Methods in Molecular Biology 268. Spencer JFT (ed). New Jersey: Humana Press. p. 491-502.
HO CS, ROCHELLE ES, MARTENS GW, SCHOOK LB, SMITH DM. 2006. Characterization of swine leukocyte antigen polymorphism by sequence-based and PCR-SSP methods in Meishan pigs. Immunogenetics 58:873-882.
IVANOSKA D, SUN DC, LUNNEY JK. 1991. Production of monoclonal antibodies reacting with polymorphic and monomorphic determinants of SLA class I expression. Immunogenetics 59: 377-89.
JONACH B, BOYE M, STOCKMARR A, JENSEN T. 2014. Fluorescence in situ hybridization of potentially pathogenic bacteria involved in neonatal porcine diarrhea. BMC Vet Res 10: 68.
LIU L, WANG J, ZHAO QH, ZI C, WU ZC, SU XM, HUO YJ, ZHU GQ, WU SL, BAO WB. 2013. Genetic variation in exon 10 of the BPI gene is associated with Escherichia coli F18 susceptibility in Sutai piglets. Gene 523: 70-75.
LUNNEY JK, HO CS, WYSOCKI M, SMITH DM. 2009. Molecular genetics of the swine major histocompatibility complex the SLA complex. Dev Comp Immunol 33: 367-374.
MADDOX-HYTTEL C, LANGKJAER RB, ENEMAR HL, VIGRE H. 2006. Cryptosporidium and Giardia in different age groups of Danish cattle and pigs- occurrence and management associated risk factors. Vet Parasitol 141: 48-59.
MORENO A, MORERA L, GARRIDO JJ, JIMÉNEZ A, ESTESO G, PAÑOS G, YUBERO N, BARBANCHO M, LLANES D. 2003. Leukocyte antigens as candidate genes to improve the immunoresponse in swine. Arch Zootec 52: 249-253.
MORRIS RS, DAVIES PR, LAWTON DE. 2002. Evolution of diseases in the world’s pig industry; Proceedings of the 17th International Pig Veterinary Society Congress; Ames, IA, USA. 2-5 Jun 2002.
NEEFJES J, JONGSMA ML, PAUL P, BAKKE O. 2011. Towards a systems understanding of MHC class I and MHC class II antigen presentation. Nat Rev Immunol 11: 823-36.
NINO-SOTO M, JAFARI JOZANI R, BRIDLE B, MALLARD BA. 2008. Analysis of gene expression patterns by microarray hybridization in blood mononuclear cells of SLA-DRB1 defined Canadian Yorkshire pigs. BMC Res Notes 1: 31.
PEDERSEN LE, HARNDAHL M, RASMUSSEN M, LAMBERTH K, GOLDE WT, LUND O, NIELSEN M, BUUS S. 2011. Porcine major histocompatibility complex (MHC) class I molecules and analysis of their peptide-binding specificities. Immunogenetics 63(12): 821-34.
PEDERSEN LE, JUNGERSEN G, SORENSEN MR, HO CS, VADEKÆR DF. 2014. Swine Leukocyte Antigen (SLA) class I allele typing of Danish swine herds and identification of commonly occurring haplotypes using sequence specific low and high resolution primers. Vet Immunol Immunopathol 162:108-116.
PENN DJ. 2002. Major Histocompatibility Complex. Encyclopedia of Life Sciences. Macmillan Publishers Ltd., Nature Publishing Group.
RENARD C, VAIMAN M, CHIANNIKULCHAI N, CATTOLICO L, ROBERT C, CHARDON P. 2001. Sequence of the pig major histocompatibility region containing the classical class I genes. Immunogenetics 53: 490-500.
REYES LM, BLOSSER RJ, SMITH RF, MINER AC, PARIS LL, BLANKENSHIP RL, TECTOR MF, TECTOR AJ. 2014. Characterization of swine leucocyte antigen alleles in a crossbred pig to be used in xenotransplant studies. Tissue Antigens 84: 484-488.
SONG D, PARK B. 2012. Porcine epidemic diarrhoea virus: a comprehensive review of molecular epidemiology, diagnosis, and vaccines. Virus Genes 44: 167-175.
STRIETER RM, BELPERIO JA, KEANE MP. 2002. Cytokines in innate host defense in the lung. J Clin Invest 109(6): 699-705.
TAJIMA S, IKAWA Y, AIDA Y. 1998. Complete bovine leukemia virus (BLV) provirus is conserved in BLV-infected cattle throughout the course of B-cell lymphosarcoma development. J Virol 113: 55-56.
TENNANT LM, RENARD C, CHARDON P, POWELL PP. 2007. Regulation of porcine classical and non-classical MHC class I expression. Immunogenetics 59: 377-389.
YE L, ZI C, PAN ZY, ZHU J, DU ZD, ZHU GQ, HUANG XG, BAO WB, WU SL. 2012. Investigation of the relationship between SLA-1 and SLA-3 gene expression and susceptibility to Escherichia coli F18 in post-weaning pigs. Comp Immunol Microbiol Infect Dis 35(1): 23-30.