Zk. Peng et al., HETEROGENEITY OF POLYCLONAL IGE CHARACTERIZED BY DIFFERENTIAL CHARGE,AFFINITY TO PROTEIN-A, AND ANTIGENICITY, Journal of allergy and clinical immunology, 100(1), 1997, pp. 87-95
Functional and physical heterogeneity of polyclonal IgE has been repor
ted. Extremely low serum concentrations of IgE have limited the study
of these important differences. We have purified polyclonal dog IgE an
d developed polyclonal and monoclonal (mAb C2) anti-dog IgE antibodies
. In this study chromatofocusing of dog IgE revealed two biologically
active IgE fractions: IgE1 eluted at pH 5.0, and IgE2 eluted at pH 4.7
. The two IgE subforms (IgEs) exhibited typical IgE characteristics: p
ositive in the 48-hour passive cutaneous anaphylaxis response, heat-la
bile, identical molecular weight, and reactive to polyclonal anti-dog
IgE. However, the two IgEs were found to be significantly heterogeneou
s. IgE1 bound to protein A and did not react with mAb C2 in ELISA and
isoelectric focusing-immunoblotting, whereas IgE2 did not bind to prot
ein A and reacted with mAb C2. Further, in sodium dodecylsulfate-polya
crylamide gel electrophoresis and immunoblotting, IgE2, but not IgE1,
reacted with seven well-defined mAb anti-human IgE antibodies and an m
Ab anti-mouse IgE antibody, even though both IgE1 and IgE2 reacted wit
h polyclonal anti-human and anti-mouse IgE. Neuraminidase or endoglyco
sidase treatment did not abolish the differential antigenicity and cha
rge of IgE1 and IgE2, although the antigenicity of IgE2 was significan
tly reduced after incubation with endoglycosidase. These data suggest
that carbohydrate moieties are not involved in the observed difference
s in antigenicity and charge and that the two IgE molecules represent
distinct isotypes. In studies with seven purified IgE fractions obtain
ed from different ragweed-allergic dogs, the distribution of ragweed I
gE2 varied 200-fold, whereas ragweed total IgE levels varied only four
fold. This raises the possibility of a relationship between different
IgEs and the allergic response.