title: Cytokine gene expression profiles of bovine dendritic cells after interaction with Mycobacterium avium ssp. paratuberculosis (M.a.p.), Escherichia coli (E. coli) or recombinant M.a.p. heat shock protein 70.
abstract: Mycobacterium avium paratuberculosis (M.a.p.) resides and replicates in macrophages. Many of the of immune mechanisms aiding M.a.p. survival in the host's cells are known. However, little is known about interactions of M.a.p. with dendritic cells (DC). As DC are important for the induction of protective immunity against infectious diseases, we investigated the interaction of M.a.p. with these cells. Quantitative real-time PCR (RT-PCR) was used to analyse differential expression of cytokine genes after 6 h and 24 h of incubation by immature DC that phagocytosed either M.a.p. or Escherichia coli (E. coli). We hypothesized that phagocytosis of E. coli would induce pro-inflammatory cytokines due to abundant presence of lipopolysaccharide (LPS) and that the cytokine expression profile induced by phagocytosis of live M.a.p. would differ. In addition we hypothesized that incubation of immature DC with rHsp70, an immunodominant antigen of M.a.p., would induce a similar profile of cytokine gene expression as phagocytosis of intact M.a.p. However, phagocytosis of both E. coli and M.a.p. resulted in a cytokine gene expression pattern representative of a (pro-)inflammatory reaction, dominated by strong induction of IL-12 gene expression, that was higher after 24 h than after 6 h of incubation, although the response to M.a.p. was less vigorous than to E. coli. Incubation with rHsp70 resulted in a more inhibitory type of cytokine gene expression, with delayed IL-12 gene expression and downregulation of the genes for IL-1beta and IL-6 after 24 h of incubation. We conclude that bovine DC produce an immuno-stimulatory, anti-mycobacterial response to infection with M.a.p., while Hsp70 potentially contributes to pathogen virulence by allowing the bacteria to invade the host cell.
authors.affiliation: Division of Immunology, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584CL Utrecht, P.O. Box 80165, 3508 TD Utrecht, The Netherlands. m.f.m.langelaar@vet.uu.nl
authors.forename: Merel F M
authors.initials: MF
authors.lastname: Langelaar
authors.forename: Corinna N
authors.initials: CN
authors.lastname: Weber
authors.forename: Marije B
authors.initials: MB
authors.lastname: Overdijk
authors.forename: Kerstin E
authors.initials: KE
authors.lastname: Müller
authors.forename: Ad P
authors.initials: AP
authors.lastname: Koets
authors.forename: Victor P M G
authors.initials: VP
authors.lastname: Rutten
mesh_terms: D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000276:immunology* / Q000382:microbiology; D002418:Cattle Diseases / Q000276:immunology; D016207:Cytokines / Q000235:genetics*; D004247:DNA / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology* / Q000382:microbiology*; D004926:Escherichia coli / Q000276:immunology*; D020869:Gene Expression Profiling; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology*; D066298:In Vitro Techniques; D016753:Interleukin-10 / Q000235:genetics; D018664:Interleukin-12 / Q000235:genetics; D016927:Mycobacterium avium subsp. paratuberculosis / Q000276:immunology* / Q000472:pathogenicity; D010283:Paratuberculosis / Q000276:immunology; D010587:Phagocytosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
chemical_list: D016207:Cytokines; D018840:HSP70 Heat-Shock Proteins; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D018664:Interleukin-12; D004247:DNA