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. 1986 Nov 1;164(5):1566–1580. doi: 10.1084/jem.164.5.1566

Idiotype variant cell populations in patients with B cell lymphoma

PMCID: PMC2188459  PMID: 3490533

Abstract

Using isolated idiotype (Id) protein we generated panels of antibodies in two patients with follicular lymphoma, one of whom had never received prior chemo-or radiotherapy. Flow cytometry and frozen section tissue staining of tumor with these monoclonal antibodies (mAb) revealed multiple subpopulations within each tumor. Individual mAb stained between 7% and 83% of surface Ig+ cells in the tumor samples. These subpopulations were overlapping and no single antibody recognized all the tumor cells. However, combinations of antibodies seemed to capture total tumor in both cases. In some instances, the percentage of tumor stained by a single mAb varied over time, and differed between lymph nodes sampled at the same time. Because a single species of Id protein was used to generate mAb in each case, it appears that the antibodies were directed against idiotopes variably shared by different populations within each tumor, and this was confirmed by crossblocking studies. Tumor cells from one patient were fused to a nonsecreting heteromyeloma line K6H6/B5, and most of the resulting hybrids secreted Id protein. Four mAb were used to screen the Id proteins secreted by these hybrids, and 11 different variants (16 maximal) were found. Southern blot analysis of rearranged Ig genes was done in two hybrids and biopsy material. Identically rearranged light-chain genes were seen but it appeared as though extensive somatic variation had occurred in heavy chain genes. These studies indicate that: striking Id variation can exist at diagnosis in untreated patients, the percentage of tumor represented by an individual variant may change with time and may differ between tumor sampled from different anatomical locations, and somatic variation appears to be responsible for the observed heterogeneity. Although this degree of variation makes anti-Id antibody therapy more difficult, appropriate combinations of mAb should be more efficacious than single antibodies in such cases.

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Selected References

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  1. Baltimore D. Somatic mutation gains its place among the generators of diversity. Cell. 1981 Nov;26(3 Pt 1):295–296. doi: 10.1016/0092-8674(81)90196-3. [DOI] [PubMed] [Google Scholar]
  2. Berek C., Griffiths G. M., Milstein C. Molecular events during maturation of the immune response to oxazolone. Nature. 1985 Aug 1;316(6027):412–418. doi: 10.1038/316412a0. [DOI] [PubMed] [Google Scholar]
  3. Brown S., Dilley J., Levy R. Immunoglobulin secretion by mouse X human hybridomas: an approach for the production of anti-idiotype reagents useful in monitoring patients with B cell lymphoma. J Immunol. 1980 Sep;125(3):1037–1043. [PubMed] [Google Scholar]
  4. Carroll W. L., Thielemans K., Dilley J., Levy R. Mouse x human heterohybridomas as fusion partners with human B cell tumors. J Immunol Methods. 1986 May 1;89(1):61–72. doi: 10.1016/0022-1759(86)90032-3. [DOI] [PubMed] [Google Scholar]
  5. Clarke S. H., Huppi K., Ruezinsky D., Staudt L., Gerhard W., Weigert M. Inter- and intraclonal diversity in the antibody response to influenza hemagglutinin. J Exp Med. 1985 Apr 1;161(4):687–704. doi: 10.1084/jem.161.4.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cleary M. L., Meeker T. C., Levy S., Lee E., Trela M., Sklar J., Levy R. Clustering of extensive somatic mutations in the variable region of an immunoglobulin heavy chain gene from a human B cell lymphoma. Cell. 1986 Jan 17;44(1):97–106. doi: 10.1016/0092-8674(86)90488-5. [DOI] [PubMed] [Google Scholar]
  7. Early P., Huang H., Davis M., Calame K., Hood L. An immunoglobulin heavy chain variable region gene is generated from three segments of DNA: VH, D and JH. Cell. 1980 Apr;19(4):981–992. doi: 10.1016/0092-8674(80)90089-6. [DOI] [PubMed] [Google Scholar]
  8. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  9. Griffiths G. M., Berek C., Kaartinen M., Milstein C. Somatic mutation and the maturation of immune response to 2-phenyl oxazolone. Nature. 1984 Nov 15;312(5991):271–275. doi: 10.1038/312271a0. [DOI] [PubMed] [Google Scholar]
  10. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  11. Haughton G., Lanier L. L., Babcock G. F., Lynes M. A. Antigen-induced murine B cell lymphomas. II. Exploitation of the surface idiotype as tumor specific antigen. J Immunol. 1978 Dec;121(6):2358–2362. [PubMed] [Google Scholar]
  12. Hieter P. A., Hollis G. F., Korsmeyer S. J., Waldmann T. A., Leder P. Clustered arrangement of immunoglobulin lambda constant region genes in man. Nature. 1981 Dec 10;294(5841):536–540. doi: 10.1038/294536a0. [DOI] [PubMed] [Google Scholar]
  13. Hozumi N., Tonegawa S. Evidence for somatic rearrangement of immunoglobulin genes coding for variable and constant regions. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3628–3632. doi: 10.1073/pnas.73.10.3628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kurosawa Y., Tonegawa S. Organization, structure, and assembly of immunoglobulin heavy chain diversity DNA segments. J Exp Med. 1982 Jan 1;155(1):201–218. doi: 10.1084/jem.155.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Levy R., Dilley J. Rescue of immunoglobulin secretion from human neoplastic lymphoid cells by somatic cell hybridization. Proc Natl Acad Sci U S A. 1978 May;75(5):2411–2415. doi: 10.1073/pnas.75.5.2411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Max E. E., Seidman J. G., Leder P. Sequences of five potential recombination sites encoded close to an immunoglobulin kappa constant region gene. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3450–3454. doi: 10.1073/pnas.76.7.3450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mayumi M., Kubagawa H., Omura G. A., Gathings W. E., Kearney J. F., Cooper M. D. Studies on the clonal origin of human B cell leukemia using monoclonal anti-idiotype antibodies. J Immunol. 1982 Aug;129(2):904–910. [PubMed] [Google Scholar]
  18. Meeker T. C., Lowder J., Maloney D. G., Miller R. A., Thielemans K., Warnke R., Levy R. A clinical trial of anti-idiotype therapy for B cell malignancy. Blood. 1985 Jun;65(6):1349–1363. [PubMed] [Google Scholar]
  19. Meeker T., Lowder J., Cleary M. L., Stewart S., Warnke R., Sklar J., Levy R. Emergence of idiotype variants during treatment of B-cell lymphoma with anti-idiotype antibodies. N Engl J Med. 1985 Jun 27;312(26):1658–1665. doi: 10.1056/NEJM198506273122602. [DOI] [PubMed] [Google Scholar]
  20. Miller R. A., Maloney D. G., Warnke R., Levy R. Treatment of B-cell lymphoma with monoclonal anti-idiotype antibody. N Engl J Med. 1982 Mar 4;306(9):517–522. doi: 10.1056/NEJM198203043060906. [DOI] [PubMed] [Google Scholar]
  21. Raffeld M., Neckers L., Longo D. L., Cossman J. Spontaneous alteration of idiotype in a monoclonal B-cell lymphoma. Escape from detection by anti-idiotype. N Engl J Med. 1985 Jun 27;312(26):1653–1658. doi: 10.1056/NEJM198506273122601. [DOI] [PubMed] [Google Scholar]
  22. Ravetch J. V., Siebenlist U., Korsmeyer S., Waldmann T., Leder P. Structure of the human immunoglobulin mu locus: characterization of embryonic and rearranged J and D genes. Cell. 1981 Dec;27(3 Pt 2):583–591. doi: 10.1016/0092-8674(81)90400-1. [DOI] [PubMed] [Google Scholar]
  23. Sakano H., Hüppi K., Heinrich G., Tonegawa S. Sequences at the somatic recombination sites of immunoglobulin light-chain genes. Nature. 1979 Jul 26;280(5720):288–294. doi: 10.1038/280288a0. [DOI] [PubMed] [Google Scholar]
  24. Siegelman M. H., Cleary M. L., Warnke R., Sklar J. Frequent biclonality and Ig gene alterations among B cell lymphomas that show multiple histologic forms. J Exp Med. 1985 Apr 1;161(4):850–863. doi: 10.1084/jem.161.4.850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  26. Stevenson G. T., Elliott E. V., Stevenson F. K. Idiotypic determinants on the surface immunoglobulin of neoplastic lymphocytes: a therapeutic target. Fed Proc. 1977 Aug;36(9):2268–2271. [PubMed] [Google Scholar]
  27. Taub R. A., Hollis G. F., Hieter P. A., Korsmeyer S., Waldmann T. A., Leder P. Variable amplification of immunoglobulin lambda light-chain genes in human populations. Nature. 1983 Jul 14;304(5922):172–174. doi: 10.1038/304172a0. [DOI] [PubMed] [Google Scholar]
  28. Teillaud J. L., Desaymard C., Giusti A. M., Haseltine B., Pollock R. R., Yelton D. E., Zack D. J., Scharff M. D. Monoclonal antibodies reveal the structural basis of antibody diversity. Science. 1983 Nov 18;222(4625):721–726. doi: 10.1126/science.6356353. [DOI] [PubMed] [Google Scholar]
  29. Thielemans K., Maloney D. G., Meeker T., Fujimoto J., Doss C., Warnke R. A., Bindl J., Gralow J., Miller R. A., Levy R. Strategies for production of monoclonal anti-idiotype antibodies against human B cell lymphomas. J Immunol. 1984 Jul;133(1):495–501. [PubMed] [Google Scholar]
  30. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983 Apr 14;302(5909):575–581. doi: 10.1038/302575a0. [DOI] [PubMed] [Google Scholar]
  31. Waldmann T. A., Korsmeyer S. J., Bakhshi A., Arnold A., Kirsch I. R. NIH conference. Molecular genetic analysis of human lymphoid neoplasms. Immunoglobulin genes and the c-myc oncogene. Ann Intern Med. 1985 Apr;102(4):497–510. doi: 10.7326/0003-4819-102-4-497. [DOI] [PubMed] [Google Scholar]
  32. Weigert M., Perry R., Kelley D., Hunkapiller T., Schilling J., Hood L. The joining of V and J gene segments creates antibody diversity. Nature. 1980 Jan 31;283(5746):497–499. doi: 10.1038/283497a0. [DOI] [PubMed] [Google Scholar]
  33. Wood G. S., Warnke R. A. The immunologic phenotyping of bone marrow biopsies and aspirates: frozen section techniques. Blood. 1982 May;59(5):913–922. [PubMed] [Google Scholar]
  34. Wysocki L., Manser T., Gefter M. L. Somatic evolution of variable region structures during an immune response. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1847–1851. doi: 10.1073/pnas.83.6.1847. [DOI] [PMC free article] [PubMed] [Google Scholar]

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