PMID- 10362264
OWN - NLM
STAT- MEDLINE
DCOM- 19990714
LR  - 20190921
IS  - 0260-437X (Print)
IS  - 0260-437X (Linking)
VI  - 19
IP  - 3
DP  - 1999 May-Jun
TI  - Cytokine endpoints for the local lymph node assay: consideration of 
      interferon-gamma and interleukin 12.
PG  - 149-55
AB  - The murine local lymph node assay (LLNA) is a method for the prospective 
      identification of contact allergens. Skin sensitization potential is assessed as 
      a function of induced proliferative responses in lymph nodes draining the site of 
      topical exposure measured in situ by incorporation of radiolabelled thymidine 
      ([3H]thymidine). The results of previous investigations have demonstrated that 
      the analysis of [3H]thymidine incorporation represents a robust and reliable 
      endpoint for the LLNA for the assessment of skin sensitizing activity for strong 
      and moderate allergens and, in addition, many weaker sensitizers. The aim of the 
      current experiments was to explore the utility of the production of the cytokines 
      interferon-gamma (IFN-gamma) and interleukin 12 (IL-12) by draining lymph node 
      cells (LNC) as alternative readouts for the LLNA. Animals were exposed to a range 
      of skin sensitizers at two application concentrations. The first of these was 
      chosen on the basis of results from previous investigations to stimulate a strong 
      proliferative response (tenfold or greater increase in proliferation compared 
      with concurrent vehicle controls). The second concentration of test material in 
      each case was the amount of chemical estimated to be necessary mathematically for 
      elicitation of a stimulation index of 3 (EC3 value); the induction of a threefold 
      or greater increase in proliferation is the current criterion for a positive 
      response in the LLNA. In addition, analyses were conducted with para-aminobenzoic 
      acid (PABA), a non-sensitizing chemical shown previously not to induce LLNA 
      responses. Secretion of IFN-gamma and the p40 subunit of IL-12 by draining LNC 
      was measured by cytokine-specific enzyme-linked immunosorbent assay. In parallel 
      experiments, LNC activity was assessed as a function of [3H]thymidine 
      incorporation in situ. All the chemical allergens tested provoked robust 
      proliferative responses, with the stimulation indices recorded at both test 
      concentrations reflecting only small changes in activity compared with previously 
      recorded data. Exposure to vehicle (4:1 acetone:olive oil, AOO) alone resulted in 
      detectable, although variable, expression of both IFN-gamma and IL-12. Treatment 
      with chemical allergen in each case caused a marked increase in IFN-gamma 
      secretion, with particularly vigorous production of cytokine being stimulated 
      following exposure to oxazolone or hexyl cinnamic aldehyde. In contrast, 
      application of chemical allergens was not generally associated with elevated 
      IL-12 p40 secretion. Exposure of mice to PABA did not result in increased 
      IFN-gamma or IL-12 production compared with vehicle-treated controls. In general, 
      however, cytokine secretion did not correlate closely with the induction of LNC 
      proliferation. These data indicate that expression by allergen-activated LNC of 
      IFN-gamma or IL-12 does not provide a reliable or sufficiently sensitive endpoint 
      for the LLNA compared with [3H]thymidine incorporation in situ.
FAU - Dearman, R J
AU  - Dearman RJ
AD  - Zeneca Central Toxicology Laboratory, Macclesfield, Cheshire, UK. 
      Rebecca.Dearman@CTL.ZENECA.com
FAU - Hilton, J
AU  - Hilton J
FAU - Basketter, D A
AU  - Basketter DA
FAU - Kimber, I
AU  - Kimber I
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Appl Toxicol
JT  - Journal of applied toxicology : JAT
JID - 8109495
RN  - 0 (Allergens)
RN  - 0 (Cytokines)
RN  - 187348-17-0 (Interleukin-12)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - Allergens/pharmacology
MH  - Animals
MH  - Cell Division/drug effects
MH  - Cells, Cultured
MH  - Cytokines/*metabolism
MH  - Female
MH  - Interferon-gamma/metabolism
MH  - Interleukin-12/metabolism
MH  - Lymph Nodes/cytology/drug effects/*metabolism
MH  - Mice
MH  - Mice, Inbred CBA
EDAT- 1999/06/11 10:00
MHDA- 2000/06/20 09:00
CRDT- 1999/06/11 10:00
PHST- 1999/06/11 10:00 [pubmed]
PHST- 2000/06/20 09:00 [medline]
PHST- 1999/06/11 10:00 [entrez]
AID - 10.1002/(SICI)1099-1263(199905/06)19:3<149::AID-JAT557>3.0.CO;2-L [pii]
AID - 10.1002/(sici)1099-1263(199905/06)19:3<149::aid-jat557>3.0.co;2-l [doi]
PST - ppublish
SO  - J Appl Toxicol. 1999 May-Jun;19(3):149-55. doi: 
      10.1002/(sici)1099-1263(199905/06)19:3<149::aid-jat557>3.0.co;2-l.