title: Cytokine release does not improve the sensitivity and specificity of the direct popliteal lymph node assay.
abstract: The popliteal lymph node assay (PLNA) is being considered as a tool to predict the potential of drugs for inducing systemic autoimmune and hypersensitivity reactions. Despite the use of different technical approaches and the evaluation of over 130 compounds, the sensitivity and specificity of the PLNA are still debatable due to many false positive and negative responses. In this study, cytokine production was assessed as a possible endpoint to improve the direct (primary) PLNA. Diclofenac, imipramine, hydralazine, glafenin and minocycline were tested using the classical procedure. TH1 cytokines (IL-2 and IFN-gamma), TH2 cytokines (IL-4 and IL-5) and pro-inflammatory cytokines (IL-6, TNF-alpha, monocyte chemoattractant protein-1 (MCP-1), IL-12p70 and IL-10) were measured in the serum and in suspensions of popliteal lymph node cells of female Balb/c mice by flow cytometry 7 days after drug administration. Only diclofenac and imipramine induced a cellularity index above 5 (considered as a positive response). Of the five tested drugs, only diclofenac induced a slight increase in TH1 cytokines, but there were no effects on TH2 cytokine production whatever the drug tested. Diclofenac increased the production of pro-inflammatory cytokines, whereas the production of MCP-1 was increased by minocycline and decreased by imipramine. No changes in serum cytokine levels were evident. These results suggest that measuring cytokine release is unlikely to improve the sensitivity and specificity of the direct PLNA.
authors.affiliation: MDS Pharma Services, Les Oncins, 69210 St. Germain sur L'Arbresle, France. guillaume.ravel@mdsps.com
authors.forename: Guillaume
authors.initials: G
authors.lastname: Ravel
authors.forename: Marielle
authors.initials: M
authors.lastname: Christ
authors.forename: Françoise
authors.initials: F
authors.lastname: Horand
authors.forename: Jacques
authors.initials: J
authors.lastname: Descotes
mesh_terms: D000818:Animals; D015551:Autoimmunity / Q000187:drug effects*; D002478:Cells, Cultured; D016207:Cytokines / Q000032:analysis / Q000378:metabolism*; D004342:Drug Hypersensitivity / Q000473:pathology*; D005260:Female; D005434:Flow Cytometry; D018836:Inflammation Mediators / Q000032:analysis / Q000378:metabolism; D023441:Local Lymph Node Assay; D051379:Mice; D008807:Mice, Inbred BALB C; D018417:Th1 Cells / Q000187:drug effects / Q000378:metabolism; D018418:Th2 Cells / Q000187:drug effects / Q000378:metabolism
chemical_list: D016207:Cytokines; D018836:Inflammation Mediators