Anti-oxazolone hybridomas and the structure of the oxazolone idiotype - PubMed (original) (raw)
. 1983 Feb;130(2):937-45.
- PMID: 6401310
Anti-oxazolone hybridomas and the structure of the oxazolone idiotype
M Kaartinen et al. J Immunol. 1983 Feb.
Abstract
Antibodies raised in several mouse and rat strains against the hapten 2-phenyloxazolone (phOx, "oxazolone") regularly contain a fraction recognized by antiidiotypic reagents. We have studied this response in BALB/c and DBA/2 mice by generating over fifty anti-phOx antibody-secreting hybridoma clones. The hybridization was performed either 7 or 14 days after a primary immunization with phOx-protein conjugate. Most of the hybrids secreted IgG1. Whereas over 80% (17/21) of IgG-producing hybrids from day-7 fusions secreted oxazolone-idiotype positive immunoglobulin, all hybridomas originating from day-14 fusions were idiotype negative. The mRNA for heavy (H) and light (L) chains of three idiotype-positive and one idiotype-negative IgG1 hybridomas were sequenced by a modification of Sanger's dideoxynucleotide method of DNA sequencing, using crude mRNA as template, synthetic oligonucleotides as primers, and reverse transcriptase to incorporate both dideoxynucleotides and labeled deoxynucleotides. The sequence of the mRNA coding for the whole variable region of each chain was established using primers complementary to the constant region near the V-C boundary and another two that coded for a framework segment in either VH or VL. This method not only provided more information than protein sequencing but was also faster and simpler. The mRNA preparation did not need fractionation beyond the poly A-containing fraction. The sequences of the H and L chain mRNA of the three idiotype-positive anti-oxazolone antibodies were extremely similar or identical, and from them a tentative oxazolone-idiotype basic sequence was derived. Only three nucleotide differences were detected; these occurred in the D segment of one H chain mRNA, in the V-J boundary of one of the light chain mRNA, and in the first hypervariable region of another. The idiotype-negative antibody had a totally different H chain mRNA and a light chain mRNA that differed by 21 bases, almost all affecting the amino acid sequence.
Similar articles
- V genes of the primary antibody response of C57BL/10 mice to the hapten phenyloxazolone.
Kaartinen M, Pelkonen E, Even J, Mäkelä O. Kaartinen M, et al. Eur J Immunol. 1988 Jul;18(7):1095-100. doi: 10.1002/eji.1830180719. Eur J Immunol. 1988. PMID: 3136024 - Inherited chi chain polymorphism in mouse antibodies to phenyloxazolone.
Karjalainen K, Hurme M, Mäkelä O. Karjalainen K, et al. Eur J Immunol. 1979 Nov;9(11):910-2. doi: 10.1002/eji.1830091114. Eur J Immunol. 1979. PMID: 118886 - Syngeneic antiidiotypic immune responses to a B cell lymphoma. Comparison between heavy chain hypervariable region peptides and intact Ig as immunogens.
Thielemans K, Rothbard JB, Levy S, Levy R. Thielemans K, et al. J Exp Med. 1985 Jul 1;162(1):19-34. doi: 10.1084/jem.162.1.19. J Exp Med. 1985. PMID: 3925067 Free PMC article. Review. - Structural implications of somatic mutations during the immune response to 2-phenyloxazolone.
Spinelli S, Alzari PM. Spinelli S, et al. Res Immunol. 1994 Jan;145(1):41-5. doi: 10.1016/s0923-2494(94)80041-3. Res Immunol. 1994. PMID: 7516564 Review. No abstract available.
Cited by
- Two Distinct Pathways in Mice Generate Antinuclear Antigen-Reactive B Cell Repertoires.
Faderl M, Klein F, Wirz OF, Heiler S, Albertí-Servera L, Engdahl C, Andersson J, Rolink A. Faderl M, et al. Front Immunol. 2018 Jan 22;9:16. doi: 10.3389/fimmu.2018.00016. eCollection 2018. Front Immunol. 2018. PMID: 29403498 Free PMC article. - Somatic hypermutation shapes the antibody repertoire of memory B cells in humans.
Meffre E, Catalan N, Seltz F, Fischer A, Nussenzweig MC, Durandy A. Meffre E, et al. J Exp Med. 2001 Aug 6;194(3):375-8. doi: 10.1084/jem.194.3.375. J Exp Med. 2001. PMID: 11489956 Free PMC article. - Analysis of immunoglobulin (Ig) isotype diversity and IgM/D memory in the response to phenyl-oxazolone.
White H, Gray D. White H, et al. J Exp Med. 2000 Jun 19;191(12):2209-20. doi: 10.1084/jem.191.12.2209. J Exp Med. 2000. PMID: 10859344 Free PMC article. - A role for RAD51 in the generation of immunoglobulin gene diversity in rabbits.
Barrington RA, Fasullo M, Knight KL. Barrington RA, et al. J Immunol. 1999 Jan 15;162(2):911-9. J Immunol. 1999. PMID: 9916715 Free PMC article. - Amino acid sequence diversity within the family of antibodies bearing the major antiarsonate cross-reactive idiotype of the A strain mouse.
Slaughter CA, Capra JD. Slaughter CA, et al. J Exp Med. 1983 Nov 1;158(5):1615-34. doi: 10.1084/jem.158.5.1615. J Exp Med. 1983. PMID: 6415209 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Research Materials