The response of porcine monocyte derived macrophages and dendritic cells to Salmonella Typhimurium and lipopolysaccharide

BMC Veterinary Research

Jiri Volf¹  Martin Faldyna¹  Zuzana Sekelova¹  Lenka Leva¹  Ondrej Polansky¹  Ivan Rychlik¹  Hana Stepanova¹  Kamila Kyrova¹ 

[1] Veterinary Research Institute, Hudcova 70, Brno 621 00, Czech Republic

关键词: Response;    Salmonella;    LPS;    Porcine;    Dendritic cell;    Macrophage;   

Others  :  1115034
DOI  :  10.1186/s12917-014-0244-1

 received in 2014-04-25, accepted in 2014-09-24,  发布年份 2014

【 摘 要 】

Background

Following infection and initial multiplication in the gut lumen, Salmonella Typhimurium crosses the intestinal epithelial barrier and comes into contact with cells of the host immune system. Mononuclear phagocytes which comprise macrophages and dendritic cells (DC) are of key importance for the outcome of Salmonella infection. Although macrophages and DC may differentiate from a common precursor, their capacities to process and present antigen differ significantly. In this study, we therefore compared the response of porcine macrophages and DC differentiated from peripheral blood monocytes to S. Typhimurium and one of the most potent bacterial pathogen associated molecular patterns, bacterial lipopolysaccharide. To avoid any bias, the expression was determined by protein LC-MS/MS and verified at the level of transcription by quantitative RT-PCR.

Results

Within 4 days of culture, peripheral blood monocytes differentiated into two populations with distinct morphology and expression of MHC II. Mass spectrometry identified 446 proteins in macrophages and 672 in DC. Out of these, 433 proteins were inducible in macrophages either after infection with S. Typhimurium or LPS exposure and 144 proteins were inducible in DC. The expression of the 46 most inducible proteins was verified at the level of transcription and the differential expression was confirmed in 22 of them. Out of these, 16 genes were induced in both cell types, 3 genes (VCAM1, HMOX1 and Serglycin) were significantly induced in macrophages only and OLDLR1 and CDC42 were induced exclusively in DC. Thirteen out of 22 up-regulated genes contained the NF-kappaB binding site in their promoters and could be considered as either part of the NF-kappaB feedback loop (IkappaBalpha and ISG15) or as NF-kappaB targets (IL1beta, IL1alpha, AMCF2, IL8, SOD2, CD14, CD48, OPN, OLDLR1, HMOX1 and VCAM1).

Conclusions

The difference in the response of monocyte derived macrophages and DC was quantitative rather than qualitative. Despite the similarity of the responses, compared to DC, the macrophages responded in a more pro-inflammatory fashion.

【 授权许可】

   
2014 Kyrova et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Kampelmacher EH, Edel W, Guinée PA, van Noorle Jansen LM: Experimental salmonella infections in pigs. Zentralblatt Für Veterinärmedizin Reihe B J Vet Med Ser B 1969, 16:717-724.
• [2]Reed WM, Olander HJ, Thacker HL: Studies on the pathogenesis of salmonella typhimurium and salmonella choleraesuis var kunzendorf infection in weanling pigs. Am J Vet Res 1986, 47:75-83.
• [3]Schwartz KJ: Salmonellosis. In Diseases of Swine. Edited by Straw BE, D’Allaire S, Mengeling WL, Taylor DJ. Iowa State University Press, Ames, Iowa; 1999:535-551.
• [4]Berends BR, Urlings HA, Snijders JM, Van Knapen F: Identification and quantification of risk factors in animal management and transport regarding salmonella spp. In pigs. Int J Food Microbiol 1996, 30:37-53.
• [5]Meurens F, Berri M, Auray G, Melo S, Levast B, Virlogeux-Payant I, Chevaleyre C, Gerdts V, Salmon H: Early immune response following salmonella enterica subspecies enterica serovar typhimurium infection in porcine jejunal gut loops. Vet Res 2009, 40:5.
• [6]Collado-Romero M, Arce C, Ramírez-Boo M, Carvajal A, Garrido JJ: Quantitative analysis of the immune response upon salmonella typhimurium infection along the porcine intestinal gut. Vet Res 2010, 41:23.
• [7]Martins RP, Collado-Romero M, Arce C, Lucena C, Carvajal A, Garrido JJ: Exploring the immune response of porcine mesenteric lymph nodes to salmonella enterica serovar typhimurium: an analysis of transcriptional changes, morphological alterations and pathogen burden. Comp Immunol Microbiol Infect Dis 2013, 36:149-160.
• [8]Rosenberger CM, Finlay BB: Phagocyte sabotage: disruption of macrophage signalling by bacterial pathogens. Nat Rev Mol Cell Biol 2003, 4:385-396.
• [9]Schnare M, Barton GM, Holt AC, Takeda K, Akira S, Medzhitov R: Toll-like receptors control activation of adaptive immune responses. Nat Immunol 2001, 2:947-950.
• [10]Carrasco CP, Rigden RC, Schaffner R, Gerber H, Neuhaus V, Inumaru S, Takamatsu H, Bertoni G, McCullough KC, Summerfield A: Porcine dendritic cells generated in vitro: morphological, phenotypic and functional properties. Immunology 2001, 104:175-184.
• [11]Auray G, Facci MR, van Kessel J, Buchanan R, Babiuk LA, Gerdts V: Differential activation and maturation of two porcine DC populations following TLR ligand stimulation. Mol Immunol 2010, 47:2103-2111.
• [12]Facci MR, Auray G, Buchanan R, van Kessel J, Thompson DR, Mackenzie-Dyck S, Babiuk LA, Gerdts V: A comparison between isolated blood dendritic cells and monocyte-derived dendritic cells in pigs. Immunology 2010, 129:396-405.
• [13]Raymond CR, Wilkie BN: Toll-like receptor, MHC II, B7 and cytokine expression by porcine monocytes and monocyte-derived dendritic cells in response to microbial pathogen-associated molecular patterns. Vet Immunol Immunopathol 2005, 107:235-247.
• [14]Sun SC, Ganchi PA, Ballard DW, Greene WC: NF-kappa B controls expression of inhibitor I kappa B alpha: evidence for an inducible autoregulatory pathway. Science 1993, 259:1912-1915.
• [15]Kapetanovic R, Fairbairn L, Beraldi D, Sester DP, Archibald AL, Tuggle CK, Hume DA: Pig bone marrow-derived macrophages resemble human macrophages in their response to bacterial lipopolysaccharide. J Immunol Baltim Md 1950 2012, 188:3382-3394.
• [16]Guzylack-Piriou L, Alves MP, McCullough KC, Summerfield A: Porcine Flt3 ligand and its receptor: generation of dendritic cells and identification of a new marker for porcine dendritic cells. Dev Comp Immunol 2010, 34:455-464.
• [17]Islam MA, Pröll M, Hölker M, Tholen E, Tesfaye D, Looft C, Schellander K, Cinar MU: Alveolar macrophage phagocytic activity is enhanced with LPS priming, and combined stimulation of LPS and lipoteichoic acid synergistically induce pro-inflammatory cytokines in pigs. Innate Immun 2013, 19:631-643.
• [18]Minakawa M, Sone T, Takeuchi T, Yokosawa H: Regulation of the nuclear factor (NF)-kappaB pathway by ISGylation. Biol Pharm Bull 2008, 31:2223-2227.
• [19]Hiscott J, Marois J, Garoufalis J, D’Addario M, Roulston A, Kwan I, Pepin N, Lacoste J, Nguyen H, Bensi G: Characterization of a functional NF-kappa B site in the human interleukin 1 beta promoter: evidence for a positive autoregulatory loop. Mol Cell Biol 1993, 13:6231-6240.
• [20]Mori N, Prager D: Transactivation of the interleukin-1alpha promoter by human T-cell leukemia virus type I and type II Tax proteins. Blood 1996, 87:3410-3417.
• [21]Keates AC, Keates S, Kwon JH, Arseneau KO, Law DJ, Bai L, Merchant JL, Wang TC, Kelly CP: ZBP-89, Sp1, and nuclear factor-kappa B regulate epithelial neutrophil-activating peptide-78 gene expression in Caco-2 human colonic epithelial cells. J Biol Chem 2001, 276:43713-43722.
• [22]Kunsch C, Rosen CA: NF-kappa B subunit-specific regulation of the interleukin-8 promoter. Mol Cell Biol 1993, 13:6137-6146.
• [23]Xu Y, Kiningham KK, Devalaraja MN, Yeh CC, Majima H, Kasarskis EJ, St Clair DK: An intronic NF-kappaB element is essential for induction of the human manganese superoxide dismutase gene by tumor necrosis factor-alpha and interleukin-1beta. DNA Cell Biol 1999, 18:709-722.
• [24]Schumann RR: Mechanisms of transcriptional activation of lipopolysaccharide binding protein (LBP). Prog Clin Biol Res 1995, 392:297-304.
• [25]Klaman LD, Thorley-Lawson DA: Characterization of the CD48 gene demonstrates a positive element that is specific to epstein-barr virus-immortalized B-cell lines and contains an essential NF-kappa B site. J Virol 1995, 69:871-881.
• [26]Samant RS, Clark DW, Fillmore RA, Cicek M, Metge BJ, Chandramouli KH, Chambers AF, Casey G, Welch DR, Shevde LA: Breast cancer metastasis suppressor 1 (BRMS1) inhibits osteopontin transcription by abrogating NF-kappaB activation. Mol Cancer 2007, 6:6. BioMed Central Full Text
• [27]Nagase M, Abe J, Takahashi K, Ando J, Hirose S, Fujita T: Genomic organization and regulation of expression of the lectin-like oxidized low-density lipoprotein receptor (LOX-1) gene. J Biol Chem 1998, 273:33702-33707.
• [28]Lavrovsky Y, Schwartzman ML, Levere RD, Kappas A, Abraham NG: Identification of binding sites for transcription factors NF-kappa B and AP-2 in the promoter region of the human heme oxygenase 1 gene. Proc Natl Acad Sci U S A 1994, 91:5987-5991.
• [29]Aderem A: The role of myristoylated protein kinase C substrates in intracellular signaling pathways in macrophages. Curr Top Microbiol Immunol 1992, 181:189-207.
• [30]Blackshear PJ: The MARCKS family of cellular protein kinase C substrates. J Biol Chem 1993, 268:1501-1504.
• [31]Yue L, Lu S, Garces J, Jin T, Li J: Protein kinase C-regulated dynamitin-macrophage-enriched myristoylated alanine-rice C kinase substrate interaction is involved in macrophage cell spreading. J Biol Chem 2000, 275:23948-23956.
• [32]Wu M, Chen DF, Sasaoka T, Tonegawa S: Neural tube defects and abnormal brain development in F52-deficient mice. Proc Natl Acad Sci U S A 1996, 93:2110-2115.
• [33]Chen J, Chang S, Duncan SA, Okano HJ, Fishell G, Aderem A: Disruption of the MacMARCKS gene prevents cranial neural tube closure and results in anencephaly. Proc Natl Acad Sci U S A 1996, 93:6275-6279.
• [34]Seykora JT, Ravetch JV, Aderem A: Cloning and molecular characterization of the murine macrophage “68-kDa” protein kinase C substrate and its regulation by bacterial lipopolysaccharide. Proc Natl Acad Sci U S A 1991, 88:2505-2509.
• [35]Sunohara JR, Ridgway ND, Cook HW, Byers DM: Regulation of MARCKS and MARCKS-related protein expression in BV-2 microglial cells in response to lipopolysaccharide. J Neurochem 2001, 78:664-672.
• [36]Rosé SD, Byers DM, Morash SC, Fedoroff S, Cook HW: Lipopolysaccharide stimulates differential expression of myristoylated protein kinase C substrates in murine microglia. J Neurosci Res 1996, 44:235-242.
• [37]Chang S, Stacey KJ, Chen J, Costelloe EO, Aderem A, Hume DA: Mechanisms of regulation of the MacMARCKS gene in macrophages by bacterial lipopolysaccharide. J Leukoc Biol 1999, 66:528-534.
• [38]Mancek-Keber M, Bencina M, Japelj B, Panter G, Andrä J, Brandenburg K, Triantafilou M, Triantafilou K, Jerala R: MARCKS as a negative regulator of lipopolysaccharide signaling. J Immunol Baltim Md 1950 2012, 188:3893-3902.
• [39]Wang X, Eaton M, Mayer M, Li H, He D, Nelson E, Christopher-Hennings J: Porcine reproductive and respiratory syndrome virus productively infects monocyte-derived dendritic cells and compromises their antigen-presenting ability. Arch Virol 2007, 152:289-303.
• [40]Stepanova H, Pavlova B, Stromerova N, Ondrackova P, Stejskal K, Slana I, Zdrahal Z, Pavlik I, Faldyna M: Different immune response of pigs to Mycobacterium avium subsp. avium and Mycobacterium avium subsp. hominissuis infection. Vet Microbiol 2012, 159:343-350.
• [41]Pavlova B, Volf J, Ondrackova P, Matiasovic J, Stepanova H, Crhanova M, Karasova D, Faldyna M, Rychlik I: SPI-1-encoded type III secretion system of Salmonella enterica is required for the suppression of porcine alveolar macrophage cytokine expression. Vet Res 2011, 42:16. BioMed Central Full Text
• [42]Wiśniewski JR, Zougman A, Nagaraj N, Mann M: Universal sample preparation method for proteome analysis. Nat Methods 2009, 6:359-362.
• [43]Boersema PJ, Raijmakers R, Lemeer S, Mohammed S, Heck AJR: Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics. Nat Protoc 2009, 4:484-494.
• [44]Wheelan SJ, Church DM, Ostell JM: Spidey: a tool for mRNA-to-genomic alignments. Genome Res 2001, 11:1952-1957.
• [45]Koressaar T, Remm M: Enhancements and modifications of primer design program Primer3. Bioinforma Oxf Engl 2007, 23:1289-1291.
• [46]Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002, 3(7):RESEARCH0034. BioMed Central Full Text