title: Inconsistencies between cytokine profiles, antibody responses, and respiratory hyperresponsiveness following dermal exposure to isocyanates.
abstract: Cytokine profiling of local lymph node responses has been proposed as a simple test to identify chemicals, such as low molecular weight diisocyanates, that pose a significant risk of occupational asthma. Previously, we reported cytokine messenger RNA (mRNA) profiles for dinitrochlorobenzene (DNCB) and six isocyanates: toluene diisocyanate, diphenylmethane-4,4'-diisocyanate, dicyclohexylmethane-4,4'-diisocyanate, isophorone diisocyanate, p-tolyl(mono)isocyanate, and meta-tetramethylene xylene diisocyanate. The present study was conducted to test the hypothesis that relative differences in the cytokine profile are predictive of relative differences in total serum immunoglobulin (Ig) E and respiratory responses to methacholine (Mch) following dermal exposure to the chemicals. After a preliminary experiment to determine an exposure regimen sufficient to achieve responses to Mch following dermal diisocyanate exposure, BALB/c mice received nine dermal exposures over a period of 28 days to one of six isocyanates, DNCB, or vehicle. Mice were then challenged with increasing doses of Mch and responsiveness was assessed using whole-body plethysmography. Serum antibody responses and cytokine mRNA profiles in the draining lymph node were also assessed. In separate experiments, cytokine protein assays were performed after five dermal exposures over a 14-day period. The response pattern for interleukin (IL)-4, IL-10, and IL-13 for the different isocyanates was highly reproducible as determined by RNAse protection assay, reverse transcription-PCR, or cytokine protein levels. However, the relative differences in T-helper cytokine profiles were not predictive of relative differences in either total serum IgE or respiratory responses to Mch following dermal exposure. The data suggest that the cytokine profiling approach needs to be further developed and refined before adoption and that other approaches to hazard identification should be pursued as well. Based on the weight of evidence from all the assays performed, it appears that all six isocyanates tested have some potential to cause respiratory hypersensitivity following dermal exposure.
authors.affiliation: National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711, USA. selgrade.maryjane@epa.gov
authors.forename: Maryjanek
authors.initials: M
authors.lastname: Selgrade
authors.forename: Elizabeth H
authors.initials: EH
authors.lastname: Boykin
authors.forename: Najwa
authors.initials: N
authors.lastname: Haykal-Coates
authors.forename: Michael R
authors.initials: MR
authors.lastname: Woolhiser
authors.forename: Connie
authors.initials: C
authors.lastname: Wiescinski
authors.forename: Debora L
authors.initials: DL
authors.lastname: Andrews
authors.forename: Aimen K
authors.initials: AK
authors.lastname: Farraj
authors.forename: Donald L
authors.initials: DL
authors.lastname: Doerfler
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Gavett
mesh_terms: D000279:Administration, Cutaneous; D000818:Animals; D000917:Antibody Formation / Q000187:drug effects* / Q000276:immunology; D018932:Chemokine CCL2 / Q000032:analysis / Q000276:immunology; D016207:Cytokines / Q000235:genetics* / Q000378:metabolism; D004137:Dinitrochlorobenzene / Q000008:administration & dosage / Q000276:immunology / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D007073:Immunoglobulin E / Q000097:blood; D007074:Immunoglobulin G / Q000097:blood; D019570:Inhalation Exposure; D017953:Isocyanates / Q000008:administration & dosage / Q000276:immunology / Q000494:pharmacology*; D008198:Lymph Nodes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D016210:Methacholine Chloride / Q000008:administration & dosage / Q000276:immunology / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012130:Respiratory Hypersensitivity / Q000139:chemically induced* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D018418:Th2 Cells / Q000187:drug effects / Q000276:immunology; D013997:Time Factors
chemical_list: C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D016207:Cytokines; D004137:Dinitrochlorobenzene; D007074:Immunoglobulin G; D017953:Isocyanates; D012333:RNA, Messenger; D016210:Methacholine Chloride; D007073:Immunoglobulin E