Assessment of glycosylation-dependent cell adhesion molecule 1 as a correlate of allergen-stimulated lymph node activation.
Early changes in gene expression have been identified by cDNA microarray technology. Analysis of draining auricular lymph node tissue sampled at 48 h following exposure to the potent contact allergen 2,4-dinitrofluorobenzene (DNFB) provided examples of up- and down-regulated genes, including onzin and guanylate binding protein 2, and glycosylation-dependent cell adhesion molecule 1 (GlyCAM-1), respectively. Allergen-induced changes in these three genes were confirmed in dose-response and kinetic analyses using Northern blotting and/or reverse transcription-polymerase chain reaction techniques. The results confirmed that these genes are robust and relatively sensitive markers of early changes provoked in the lymph node by contact allergen. Upon further investigation, it was found that altered expression of the adhesion molecule GlyCAM-1 was not restricted to treatment with DNFB. Topical sensitization of mice to a chemically unrelated contact allergen, oxazolone, was also associated with a decrease in the expression of mRNA for GlyCAM-1. Supplementary experiments revealed that changes in expression of this gene are independent of the stimulation by chemical allergens of proliferative responses by draining lymph node cells. Taken together these data indicate that the expression of GlyCAM-1 is down-regulated rapidly following epicutaneous treatment of mice with chemical allergens, but that this reduction is associated primarily with changes in lymph node cell number, or some other aspect of lymph node activation, rather than proliferation.
Betts
Catherine J
CJ
Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire SK10 4TJ, UK. catherine.betts@syngenta.com
Moggs
Jonathan G
JG
Caddick
Helen T
HT
Cumberbatch
Marie
M
Orphanides
George
G
Dearman
Rebecca J
RJ
Ryan
Cindy A
CA
Hulette
Ben C
BC
Frank Gerberick
G
G
Kimber
Ian
I
D000287:Administration, Topical; D000485:Allergens / Q000008:administration & dosage / Q000633:toxicity*; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D017931:DNA Primers / Q000737:chemistry; D003877:Dermatitis, Contact / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D004139:Dinitrofluorobenzene / Q000008:administration & dosage / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005260:Female; D005786:Gene Expression Regulation; D008198:Lymph Nodes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D009077:Mucins / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity*; D020411:Oligonucleotide Array Sequence Analysis; D010081:Oxazolone / Q000008:administration & dosage / Q000633:toxicity*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000485:Allergens; D017931:DNA Primers; D009077:Mucins; D012333:RNA, Messenger; C075185:sulfated glycoprotein p50; D010081:Oxazolone; D004139:Dinitrofluorobenzene