title: Evidence for active antigen presentation by monocyte/macrophages in response to stimulation with particles: the expression of NFκB transcription factors and costimulatory molecules.
abstract: BACKGROUND
  The macrophage and lymphocyte response to wear debris contributes to the failure of some joint replacements. Costimulatory molecule expression by particle-containing macrophages is an evidence for antigen presentation. The NFκB transcription factors are regulators of costimulatory molecules and are present in tissue near failed joint prostheses. The tissue localisation of NFκB and the expression of these factors and costimulatory molecules by U937 cells stimulated with nano- and microparticles are reported, together with the effects of an NFκB inhibitor (sc514).
  
  
  METHODS
  The tissue localisation of RelA, RelB, c-rel, p50, p52 and NF-IL6 was examined by immunohistochemistry in samples from 15 patients with failure of metal against polyethylene total hip replacements. The expression of these NFκB factors by U937 cells stimulated with microparticles (CoCr, diamond) and nanoparticles (diamond) was examined by quantified RT-PCR. Lipopolysaccharide provided positive controls while negative controls had no additions to culture. Inhibition of NFκB activity by sc-514 was studied. The expression of costimulatory molecules (CD80, CD86 and HLA-DR) was evaluated in parallel cell culture studies by tricolour flow cytometry.
  
  
  CONCLUSIONS
  Immunohistochemistry of tissue showed the highest expression for NF-IL6 (32.56 ± 11.61 per cent), RelA (33.66 ± 9.98 per cent) and p52 (32.07 ± 12.90 per cent), then RelB (22.63 ± 7.49 per cent), c-rel (14.07 ± 6.72 per cent) and p50 (13.07 ± 5.99 per cent). NF-IL6 was localised to macrophages, RelB to RFD1+ dendritic cells. U937 cells showed an increased expression of all NFκB factors (p < 0.01) in response to CoCr and diamond microparticles. Only RelA and c-rel (p < 0.01) were increased by one diamond nanoparticle and p52 and c-rel (p < 0.01) by another nanoparticulate diamond. Inhibition by sc-514 of RelA, c-rel and p50 expression occurred with all four particles, p52 was decreased for all diamond particles (but not CoCr) and RelB was not inhibited with any of the particles. CD86 and HLA-DR expression were upregulated by microparticles (CoCr, diamond) (p ≪ 0.01) with lower levels (significant) of these molecules found with diamond nanoparticles. CD80 expression was much less than CD86 and HLA-DR. Costimulatory molecule expression in the bone-implant interface indicates antigen presentation by macrophages. Functional studies with U937 monocytes show the same molecules expressed on exposure to micro- and nanoparticles. Highest values occur with CoCr while the smallest diamond nanoparticles are the least stimulatory. NFκB expression gives an insight into the immunogenic potential of the different particles.
authors.affiliation: Division of Surgery and Cancer, Imperial College, London SW7 2AZ, UK.
authors.forename: Huwaidha
authors.initials: H
authors.lastname: Altaf
authors.forename: Peter A
authors.initials: PA
authors.lastname: Revell
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D017951:Antigen Presentation / Q000187:drug effects* / Q000276:immunology; D018122:B7-1 Antigen / Q000235:genetics / Q000276:immunology; D051940:B7-2 Antigen / Q000235:genetics / Q000276:immunology; D002858:Chromium Alloys / Q000737:chemistry / Q000633:toxicity; D018130:Diamond / Q000737:chemistry / Q000633:toxicity; D005260:Female; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000276:immunology; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology; D006801:Humans; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology; D008297:Male; D046529:Microscopy, Electron, Transmission; D008875:Middle Aged; D009000:Monocytes / Q000187:drug effects* / Q000276:immunology; D016328:NF-kappa B / Q000235:genetics* / Q000276:immunology; D053758:Nanoparticles / Q000737:chemistry / Q000633:toxicity*; D010316:Particle Size; D011475:Prosthesis Failure / Q000009:adverse effects; D013499:Surface Properties; D020298:U937 Cells
chemical_list: D018122:B7-1 Antigen; D051940:B7-2 Antigen; C497964:CD86 protein, human; D002858:Chromium Alloys; D006684:HLA-DR Antigens; D016328:NF-kappa B; D018130:Diamond