Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1988 Jul 1;168(1):55–71. doi: 10.1084/jem.168.1.55

Retinoic acid induces the differentiation of B cell hybridomas from patients with common variable immunodeficiency

PMCID: PMC2189392  PMID: 3294336

Abstract

Human-human B cell hybridomas constructed from B lymphocytes of common variable immunodeficiency (CVI) patients and the nonsecreting cell line WIL2/729 HF consistently secrete low levels of Ig and appear to retain a defect characteristic of the CVI patient's B cells. We assessed the differentiative capacity of retinoic acid (RA) on these hybridomas, as well as on hybridomas constructed from normal B cells and from patients with selective IgA deficiency. RA at concentrations varying between 10(- 5) and 10(-9) M augmented IgM secretion 4-20-fold from four of four CVI hybridomas tested, but did not affect Ig secretion from normal or IgA- deficiency hybridomas. In support of this elevated Ig secretion, RA enhanced the de novo synthesis of biosynthetically labeled light (kappa) and heavy (mu) Ig (up to 4- and 15-fold, respectively) in the CVI hybridoma line JK32.1. The increase in IgM synthesis/secretion could not be accounted for by RA-induced alteration in the cell cycle. In inducing this increase in IgM production, RA was found to affect two aspects of Ig gene expression: (a) the steady-state levels of heavy and light chain mRNAs were enhanced, and (b) the processing of mu heavy chain transcripts to the secreted mRNA form became favored over the membrane mRNA form. We also show that expression of Leu-17 (CD38), a surface marker that is re-expressed in the late pre-plasma stage of B cell development, was increased by RA from less than 20% to greater than 90% of the total cell population, with a concomitant 4-10-fold augmentation in the mean fluorescence intensity. Changes in both Leu-17 expression and de novo Ig synthesis were prominent by 24 h, but could be observed as early as 8 h after induction. Taken together, our study demonstrates that RA affects a marked alteration in the differentiated state of the CVI hybridoma clones. This finding suggests that retinoids can enhance the functional capabilities of B cells with defects in maturation and support further studies to evaluate their clinical potential in CVI.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ashman R. F., Saxon A., Stevens R. H. Profile of multiple lymphocyte functional defects in acquired hypogrammaglobulinemia, derived from in vitro cell recombination analysis. J Allergy Clin Immunol. 1980 Apr;65(4):242–256. doi: 10.1016/0091-6749(80)90151-7. [DOI] [PubMed] [Google Scholar]
  2. Breitman T. R., Selonick S. E., Collins S. J. Induction of differentiation of the human promyelocytic leukemia cell line (HL-60) by retinoic acid. Proc Natl Acad Sci U S A. 1980 May;77(5):2936–2940. doi: 10.1073/pnas.77.5.2936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buell D. N., Fahey J. L. Limited periods of gene expression in immunoglobulin-synthesizing cells. Science. 1969 Jun 27;164(3887):1524–1525. doi: 10.1126/science.164.3887.1524. [DOI] [PubMed] [Google Scholar]
  4. Clark E. A., Shu G. Activation of human B cell proliferation through surface Bp35 (CD20) polypeptides or immunoglobulin receptors. J Immunol. 1987 Feb 1;138(3):720–725. [PubMed] [Google Scholar]
  5. Davis R. C., Thomason A. R., Fuller M. L., Slovin J. P., Chou C. C., Chada S., Gatti R. A., Salser W. A. mRNA species regulated during the differentiation of HL-60 cells to macrophages and neutrophils. Dev Biol. 1987 Jan;119(1):164–174. doi: 10.1016/0012-1606(87)90218-1. [DOI] [PubMed] [Google Scholar]
  6. Denis K. A., Wall R., Saxon A. Human-human B cell hybridomas from in vitro stimulated lymphocytes of patients with common variable immunodeficiency. J Immunol. 1983 Nov;131(5):2273–2278. [PubMed] [Google Scholar]
  7. Gerster T., Picard D., Schaffner W. During B-cell differentiation enhancer activity and transcription rate of immunoglobulin heavy chain genes are high before mRNA accumulation. Cell. 1986 Apr 11;45(1):45–52. doi: 10.1016/0092-8674(86)90536-2. [DOI] [PubMed] [Google Scholar]
  8. Hermans P. E., Diaz-Buxo J. A., Stobo J. D. Idiopathic late-onset immunoglobulin deficiency. Clinical observations in 50 patients. Am J Med. 1976 Aug;61(2):221–237. doi: 10.1016/0002-9343(76)90173-x. [DOI] [PubMed] [Google Scholar]
  9. Lacroix A., Lippman M. E. Binding of retinoids to human breast cancer cell lines and their effects on cell growth. J Clin Invest. 1980 Mar;65(3):586–591. doi: 10.1172/JCI109703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lamson G., Koshland M. E. Changes in J chain and mu chain RNA expression as a function of B cell differentiation. J Exp Med. 1984 Sep 1;160(3):877–892. doi: 10.1084/jem.160.3.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Linder S., Krondahl U., Sennerstam R., Ringertz N. R. Retinoic acid-induced differentiation of F9 embryonal carcinoma cells. Exp Cell Res. 1981 Apr;132(2):453–460. doi: 10.1016/0014-4827(81)90120-8. [DOI] [PubMed] [Google Scholar]
  13. Lotan R. Effects of vitamin A and its analogs (retinoids) on normal and neoplastic cells. Biochim Biophys Acta. 1980 Mar 12;605(1):33–91. doi: 10.1016/0304-419x(80)90021-9. [DOI] [PubMed] [Google Scholar]
  14. Matsumoto M., Tominaga A., Harada N., Takatsu K. Role of T cell-replacing factor (TRF) in the murine B cell differentiation: induction of increased levels of expression of secreted type IgM mRNA. J Immunol. 1987 Mar 15;138(6):1826–1833. [PubMed] [Google Scholar]
  15. Potter H., Weir L., Leder P. Enhancer-dependent expression of human kappa immunoglobulin genes introduced into mouse pre-B lymphocytes by electroporation. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7161–7165. doi: 10.1073/pnas.81.22.7161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Reinherz E. L., Cooper M. D., Schlossman S. F., Rosen F. S. Abnormalities of T cell maturation and regulation in human beings with immunodeficiency disorders. J Clin Invest. 1981 Sep;68(3):699–705. doi: 10.1172/JCI110305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sherr E. H., Saxon A., Wells J. R. Functional and phenotypic characterization of human B lymphocyte subsets isolated by unit gravity sedimentation. Int Arch Allergy Appl Immunol. 1988;85(2):154–160. doi: 10.1159/000234495. [DOI] [PubMed] [Google Scholar]
  18. Sherr E. H., Stein L. D., Dosch H. M., Saxon A. IgE-enhancing activity directly and selectively affects activated B cells: evidence for a human IgE differentiation factor. J Immunol. 1987 Jun 1;138(11):3836–3843. [PubMed] [Google Scholar]
  19. Sidell N., Famatiga E., Golub S. H. Immunological aspects of retinoids in humans. II. Retinoic acid enhances induction of hemolytic plaque-forming cells. Cell Immunol. 1984 Oct 15;88(2):374–381. doi: 10.1016/0008-8749(84)90170-9. [DOI] [PubMed] [Google Scholar]
  20. Sidell N. Retinoic acid-induced growth inhibition and morphologic differentiation of human neuroblastoma cells in vitro. J Natl Cancer Inst. 1982 Apr;68(4):589–596. [PubMed] [Google Scholar]
  21. Sidell N., Rieber P., Golub S. H. Immunological aspects of retinoids in humans. I. Analysis of retinoic acid enhancement of thymocyte responses to PHA. Cell Immunol. 1984 Aug;87(1):118–125. doi: 10.1016/0008-8749(84)90136-9. [DOI] [PubMed] [Google Scholar]
  22. Tedder T. F., Clement L. T., Cooper M. D. Discontinuous expression of a membrane antigen (HB-7) during B lymphocyte differentiation. Tissue Antigens. 1984 Sep;24(3):140–149. doi: 10.1111/j.1399-0039.1984.tb02118.x. [DOI] [PubMed] [Google Scholar]
  23. Thaller C., Eichele G. Identification and spatial distribution of retinoids in the developing chick limb bud. Nature. 1987 Jun 18;327(6123):625–628. doi: 10.1038/327625a0. [DOI] [PubMed] [Google Scholar]
  24. Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
  25. Waldmann T. A., Durm M., Broder S., Blackman M., Blaese R. M., Strober W. Role of suppressor T cells in pathogenesis of common variable hypogammaglobulinaemia. Lancet. 1974 Sep 14;2(7881):609–613. doi: 10.1016/s0140-6736(74)91940-0. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES