Transcriptome analysis of porcine PBMCs after in vitro stimulation by LPS or PMA/ionomycin using an expression array targeting the pig immune response.
BACKGROUND
Designing sustainable animal production systems that better balance productivity and resistance to disease is a major concern. In order to address questions related to immunity and resistance to disease in pig, it is necessary to increase knowledge on its immune system and to produce efficient tools dedicated to this species.


RESULTS
A long-oligonucleotide-based chip referred to as SLA-RI/NRSP8-13K was produced by combining a generic set with a newly designed SLA-RI set that targets all annotated loci of the pig major histocompatibility complex (MHC) region (SLA complex) in both orientations as well as immunity genes outside the SLA complex. The chip was used to study the immune response of pigs following stimulation of porcine peripheral blood mononuclear cells (PBMCs) with lipopolysaccharide (LPS) or a mixture of phorbol myristate acetate (PMA) and ionomycin for 24 hours. Transcriptome analysis revealed that ten times more genes were differentially expressed after PMA/ionomycin stimulation than after LPS stimulation. LPS stimulation induced a general inflammation response with over-expression of SAA1, pro-inflammatory chemokines IL8, CCL2, CXCL5, CXCL3, CXCL2 and CCL8 as well as genes related to oxidative processes (SOD2) and calcium pathways (S100A9 and S100A12). PMA/ionomycin stimulation induced a stronger up-regulation of T cell activation than of B cell activation with dominance toward a Th1 response, including IL2, CD69 and TNFRSF9 (tumor necrosis factor receptor superfamily, member 9) genes. In addition, a very intense repression of THBS1 (thrombospondin 1) was observed. Repression of MHC class I genes was observed after PMA/ionomycin stimulation despite an up-regulation of the gene cascade involved in peptide processing. Repression of MHC class II genes was observed after both stimulations. Our results provide preliminary data suggesting that antisense transcripts mapping to the SLA complex may have a role during immune response.


CONCLUSIONS
The SLA-RI/NRSP8-13K chip was found to accurately decipher two distinct immune response activations of PBMCs indicating that it constitutes a valuable tool to further study immunity and resistance to disease in pig. The transcriptome analysis revealed specific and common features of the immune responses depending on the stimulation agent that increase knowledge on pig immunity.
Gao
Yu
Y
INRA, UMR 1313 de Génétique Animale et Biologie Intégrative, Domaine de Vilvert, 78350 Jouy-en-Josas, France.
Flori
Laurence
L
Lecardonnel
Jérome
J
Esquerré
Diane
D
Hu
Zhi-Liang
ZL
Teillaud
Angélique
A
Lemonnier
Gaëtan
G
Lefèvre
Francois
F
Oswald
Isabelle P
IP
Rogel-Gaillard
Claire
C
D000595:Amino Acid Sequence; D000818:Animals; D020869:Gene Expression Profiling; D053263:Gene Regulatory Networks; D006649:Histocompatibility Antigens / Q000235:genetics; D015759:Ionomycin / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology* / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000276:immunology; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D034421:Sus scrofa / Q000276:immunology* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000276:immunology
D006649:Histocompatibility Antigens; D008070:Lipopolysaccharides; D015759:Ionomycin; D013755:Tetradecanoylphorbol Acetate
Genomics. 2006 Jul;88(1):96-110
16515853
Nat Rev Genet. 2009 Mar;10(3):184-94
19223927
J Immunol. 1998 Aug 1;161(3):1220-30
9686582
Clin Diagn Lab Immunol. 2004 Jul;11(4):691-8
15242943
BMC Genomics. 2005 Sep 22;6:135
16176586
Vet Immunol Immunopathol. 2002 Aug;87(1-2):11-8
12052338
Curr Opin Microbiol. 2008 Jun;11(3):251-6
18567532
BMC Genomics. 2005 May 10;6:70
15885146
Blood. 2006 Sep 1;108(5):1751-7
16735604
Proc Natl Acad Sci U S A. 2002 Jan 22;99(2):972-7
11805339
Immunogenetics. 1992;36(2):117-20
1612643
J Cell Biol. 1995 Aug;130(3):503-6
7542656
Genomics. 2005 Apr;85(4):503-11
15780753
J Leukoc Biol. 2008 Jan;83(1):139-48
17962368
BMC Genomics. 2008 Sep 23;9:437
18811943
J Biol Chem. 1994 Jan 7;269(1):77-85
8276881
Nat Rev Immunol. 2009 Apr;9(4):271-85
19319144
J Pineal Res. 2007 Sep;43(2):121-9
17645690
Nature. 2003 Aug 14;424(6950):788-93
12917688
Blood. 2008 Jan 15;111(2):613-23
17890448
Exp Lung Res. 2006 Sep;32(8):331-47
17090475
BMC Genomics. 2008 Mar 10;9:123
18331636
Med Immunol. 2002 Nov 18;1(1):2
12459040
Bioinformatics. 2003 Feb 12;19(3):368-75
12584122
Cytokine. 2008 May;42(2):145-151
18304834
Am J Physiol Heart Circ Physiol. 2006 Apr;290(4):H1433-45
16272197
BMC Genomics. 2009 Feb 05;10:64
19196461
J Immunol. 1997 May 1;158(9):4260-9
9126988
Can J Vet Res. 2007 Apr;71(2):108-18
17479774
Anim Genet. 2006 Aug;37(4):423-4
16879364
Science. 1995 Oct 20;270(5235):467-70
7569999
Exp Mol Med. 2000 Sep 30;32(3):135-40
11048644
Nat Rev Immunol. 2006 Oct;6(10):751-60
16998508
Nat Rev Microbiol. 2005 Apr;3(4):281-94
15806094
Genome Biol. 2004;5(10):R80
15461798
Vet Immunol Immunopathol. 2004 Dec 8;102(3):199-216
15507306
Curr Opin Microbiol. 2006 Apr;9(2):138-42
16459132
Genomics. 2005 Nov;86(5):618-25
16216716
Microbes Infect. 2008 Jul;10(9):1067-71
18662797
J Immunol. 2001 Jul 1;167(1):283-8
11418661
Vet Res. 2008 Nov-Dec;39(6):54
18638439
Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14863-8
9843981
Vet Immunol Immunopathol. 2004 Sep;101(1-2):1-17
15261689
J Cell Biol. 1997 Aug 11;138(3):707-17
9245797
Int J Biol Sci. 2007 Feb 09;3(3):132-52
17384733
BMC Genomics. 2006 Mar 13;7:49
16533398
Mutat Res. 2004 May 18;549(1-2):241-53
15120974
Methods. 2001 Dec;25(4):402-8
11846609
Int Immunopharmacol. 2006 Feb;6(2):133-43
16399618
J Appl Genet. 2005;46(2):171-7
15876684
J Biol Chem. 1996 Jan 5;271(1):21-4
8550562
Stat Appl Genet Mol Biol. 2004;3:Article3
16646809
Clin Microbiol Rev. 1999 Jan;12(1):147-79
9880479
J Immunol. 2004 Apr 1;172(7):4583-91
15034076
Immunol Lett. 2005 May 15;98(2):280-90
15860229
Cancer Immunol Immunother. 2009 Apr;58(4):567-74
18751977
J Virol. 2006 Nov;80(21):10514-21
17041222
BMC Genomics. 2008 Jan 14;9:17
18194535
BMC Immunol. 2003 Jun 26;4:5
12826024
Genomics. 2008 Nov;92(5):255-64
18703132
PLoS One. 2009;4(2):e4427
19212463