PMID- 15588913
OWN - NLM
STAT- MEDLINE
DCOM- 20050224
LR  - 20191210
IS  - 0300-483X (Print)
IS  - 0300-483X (Linking)
VI  - 206
IP  - 2
DP  - 2005 Jan 15
TI  - Evaluation of an in vitro method for the measurement of specific IgE antibody 
      responses: the rat basophilic leukemia (RBL) cell assay.
PG  - 195-205
AB  - The evaluation of allergenic potential is a key parameter in the safety 
      assessment of novel proteins, including those expressed in genetically modified 
      crops and foodstuffs. The majority of allergic reactions to food proteins are 
      immediate type hypersensitivity reactions in which the principal biological 
      effector is IgE antibody; the accurate measurement of specific IgE antibody is 
      therefore a critical factor in experimental systems designed to characterize 
      protein allergenic potential. Due to the presence of much higher concentrations 
      of other immunoglobulin isotypes, the assessment of specific serum IgE antibody 
      poses substantial technical challenges. We have examined the utility of the rat 
      basophilic leukemia (RBL) cell line for the measurement of murine IgE responses. 
      RBL cells were sensitized with mouse monoclonal anti-dinitrophenyl (DNP) IgE 
      antibody and challenged with DNP-albumin conjugates with various hapten 
      substitution ratios (SR). Polyclonal anti-OVA IgE antisera were also assessed for 
      activity in the RBL assay. Results were compared with titers measured in 
      homologous passive cutaneous anaphylaxis (PCA) assay. Marked degranulation of RBL 
      cells was induced by conjugates with SRs of between 16 and 32, whereas conjugates 
      with lower SRs (of 10 or 3) failed to elicit significant serotonin release. All 
      conjugates were able to induce mast cell degranulation in vivo in a PCA assay. 
      Anti-OVA antisera with PCA titers of 1/32 to 1/64 failed to stimulate RBL cell 
      degranulation, whereas high titer antibody (1/2048 to 1/4096 by PCA) induced a 
      positive RBL cell response. Successful stimulation of RBL cell degranulation 
      requires not only appropriate epitope densities but also high affinity antibody. 
      These data indicate that this assay is inappropriate for the routine analysis of 
      specific polyclonal IgE antibody responses such as those that are induced by 
      exposure to complex protein allergens.
FAU - Dearman, R J
AU  - Dearman RJ
AD  - Syngenta Central Toxicology Laboratory, Alderley Park, Macclesfield, Cheshire 
      SK10 4TJ, UK. rebecca.dearman@syngenta.com
FAU - Skinner, R A
AU  - Skinner RA
FAU - Deakin, N
AU  - Deakin N
FAU - Shaw, D
AU  - Shaw D
FAU - Kimber, I
AU  - Kimber I
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Journal Article
PL  - Ireland
TA  - Toxicology
JT  - Toxicology
JID - 0361055
RN  - 0 (Allergens)
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Dinitrobenzenes)
RN  - 0 (Serum Albumin)
RN  - 37341-29-0 (Immunoglobulin E)
RN  - 9006-59-1 (Ovalbumin)
SB  - IM
MH  - Allergens/*immunology
MH  - Animals
MH  - Antibodies, Monoclonal/immunology
MH  - Antibody Specificity
MH  - Basophils/immunology
MH  - Cell Degranulation/immunology
MH  - Cell Line, Tumor
MH  - Dinitrobenzenes/*immunology
MH  - Dose-Response Relationship, Drug
MH  - Female
MH  - Immunoglobulin E/*biosynthesis/blood/immunology
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Ovalbumin/*immunology
MH  - Passive Cutaneous Anaphylaxis
MH  - Rats
MH  - Serum Albumin/immunology
MH  - Specific Pathogen-Free Organisms
EDAT- 2004/12/14 09:00
MHDA- 2005/02/25 09:00
CRDT- 2004/12/14 09:00
PHST- 2004/05/30 00:00 [received]
PHST- 2004/08/02 00:00 [accepted]
PHST- 2004/12/14 09:00 [pubmed]
PHST- 2005/02/25 09:00 [medline]
PHST- 2004/12/14 09:00 [entrez]
AID - S0300-483X(04)00468-8 [pii]
AID - 10.1016/j.tox.2004.08.007 [doi]
PST - ppublish
SO  - Toxicology. 2005 Jan 15;206(2):195-205. doi: 10.1016/j.tox.2004.08.007.