Characterization, Expression and Genetic Analysis of Recurrent Idiotypes in the Unrestricted Anti-Fluorescein Immune Response

Eighteen monoclonal anti-fluorescein (Fl) antibodies from BALB/cV mice were characterized and 13 possessed a low affinity for Fl (K(,a) = 5 x 10('4) M('-1)) while 5 had a higher affinity (K(,a) (TBOND) 5 x 10('8) M('-1)). Eleven of the 13 low affinity clones were idiotypically cross-reactive (40-100%) and 4 of these clones possessed identical idiotypes (Ids) (3-12, 3-13, 3-17 and 3-35). Heavy and light chains derived from clones 3-12, 3-13 and 3-17 and used in recombination experiments, resulted in the reformation of an anti-Fl active site in all six heterologous combinations. The expression of Ids 3-13 and 3-17 in the I(DEGREES), II(DEGREES) or III(DEGREES) polyclonal anti-Fl response was quantitated and found to represent (LESSTHEQ) 1.0% of the specific repertoire. Ids 3-13 and 3-17 were expressed in 15/22 ad 22/28 individual mice, respectively. Clones 3-13 and 3-17 had similar though not identical active sites as determined by fine specificity of binding and Id inhibitions. Ids 3-13 and 3-17 represent the first demonstration of repertoire-specific recurrent Ids expressed during a T-dependent immune response lacking a dominant Id. Antibodies 3-13 and 3-17 were idiotypically cross-reactive (10-40%) with 6 other anti-Fl clones (designated the 9-40 family) which possessed a higher affinity for Fl (K(,a) (TURN) 5 x 10('6) M('-1)). Members of the 9-40 family were also idiotypically cross-reactive with a single high affinity clone 4-4 (K(,a) = 5 x 10('9) M('-1)). This idiotypic linkage provided a greater span in affinity (>20,000 fold) than in all previously characterized Id families within restricted or unrestricted immune responses. The sequence of antibody 3-13 V(,H) region was determined by genomic cloning and amino acid sequencing and was 78% homologous to the (alpha)-1,3-dextran binding myeloma proteins J558 and 104E. Rearrangement of a V(,H) gene homologous to 3-13 was required idiotypically cross-reactive clones and in no idiotypically distinct clones.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Microbiology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Reinitz, David Maximilian

• Biology, Microbiology

Identifier

(UMI)AAI8701597

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/71179