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. 1988 Aug 1;168(2):589–603. doi: 10.1084/jem.168.2.589

Comparison of the fetal and adult functional B cell repertoires by analysis of VH gene family expression

PMCID: PMC2189009  PMID: 3261774

Abstract

The functional B cell repertoire in BALB/c mice was assessed at various stages in ontogeny. This was done by analyzing VH gene family expression using the sensitive technique of in situ hybridization. The B cell repertoire was probed with the mitogen, LPS, and the antigen DNP. DNP was chosen because B cells responsive to this hapten appear very early in ontogeny. The APCs that developed after stimulation with LPS or DNP were analyzed for VH gene expression by in situ hybridization of individual cells using radiolabeled VH gene family probes. The results indicated that VH gene expression in fetal B cells after stimulation was distinct from adult B cells in that there was a biased expression of D proximal families. The results indicated that this bias was associated with developmental age and not a given differentiation stage in the B cell lineage. In addition, stimulation of fetal B cells with DNP resulted in a large increase in expression of member(s) of VH 36-60, suggesting that the early appearance of DNP- responsive B cells is not strictly correlated with preferential rearrangement of D proximal families, VH 7183 and VH Q52. However, the results suggested that a large proportion of pre-B cells that preferentially rearrange D proximal families early in ontogeny become part of the functional developing repertoire.

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Selected References

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  1. Abraham K. M., Teale J. M. Isotype restriction during infection of mice with the cestode Mesocestoides corti: role of immune suppression. J Immunol. 1987 Mar 15;138(6):1699–1704. [PubMed] [Google Scholar]
  2. Alt F. W., Blackwell T. K., Yancopoulos G. D. Development of the primary antibody repertoire. Science. 1987 Nov 20;238(4830):1079–1087. doi: 10.1126/science.3317825. [DOI] [PubMed] [Google Scholar]
  3. Bailey J. N., Klement J. F., McAllister W. T. Relationship between promoter structure and template specificities exhibited by the bacteriophage T3 and T7 RNA polymerases. Proc Natl Acad Sci U S A. 1983 May;80(10):2814–2818. doi: 10.1073/pnas.80.10.2814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blankenstein T., Krawinkel U. Immunoglobulin VH region genes of the mouse are organized in overlapping clusters. Eur J Immunol. 1987 Sep;17(9):1351–1357. doi: 10.1002/eji.1830170920. [DOI] [PubMed] [Google Scholar]
  5. Brodeur P. H., Riblet R. The immunoglobulin heavy chain variable region (Igh-V) locus in the mouse. I. One hundred Igh-V genes comprise seven families of homologous genes. Eur J Immunol. 1984 Oct;14(10):922–930. doi: 10.1002/eji.1830141012. [DOI] [PubMed] [Google Scholar]
  6. Cancro M. P., Wylie D. E., Gerhard W., Klinman N. R. Patterned acquisition of the antibody repertoire: diversity of the hemagglutinin-specific B-cell repertoire in neonatal BALB/c mice. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6577–6581. doi: 10.1073/pnas.76.12.6577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Denis K. A., Klinman N. R. Genetic and temporal control of neonatal antibody expression. J Exp Med. 1983 Apr 1;157(4):1170–1183. doi: 10.1084/jem.157.4.1170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dildrop R., Krawinkel U., Winter E., Rajewsky K. VH-gene expression in murine lipopolysaccharide blasts distributes over the nine known VH-gene groups and may be random. Eur J Immunol. 1985 Nov;15(11):1154–1156. doi: 10.1002/eji.1830151117. [DOI] [PubMed] [Google Scholar]
  9. Dzierzak E. A., Janeway C. A., Jr, Rosenstein R. W., Gottlieb P. D. Expression of an idiotype (Id-460) during in vivo anti-dinitrophenyl antibody responses. I. Mapping of genes for Id-460 expression to the variable region of immunoglobulin heavy-chain locus and to the variable region of immunoglobulin kappa-light-chain locus. J Exp Med. 1980 Sep 1;152(3):720–729. doi: 10.1084/jem.152.3.720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dzierzak E. A., Rosenstein R. W., Janeway C. A., Jr Expression of an idiotype (Id-460) during in vivo anti-dinitrophenyl antibody responses. II. Transient idiotypic dominance. J Exp Med. 1981 Nov 1;154(5):1432–1441. doi: 10.1084/jem.154.5.1432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Froscher B. G., Klinman N. R. Strain-specific silencing of a predominant antidextran clonotype family. J Exp Med. 1985 Nov 1;162(5):1620–1633. doi: 10.1084/jem.162.5.1620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fung J., Köhler H. Immune response to phosphorylcholine. VII. Functional evidence for three separate B cell subpopulations responding to TI and TD PC-antigens. J Immunol. 1980 Aug;125(2):640–646. [PubMed] [Google Scholar]
  13. Haas W., Layton J. E. Separation of antigen-specific lymphocytes. I. Enrichment of antigen-binding cells. J Exp Med. 1975 May 1;141(5):1004–1014. doi: 10.1084/jem.141.5.1004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hardy R. R., Hayakawa K. Development and physiology of Ly-1 B and its human homolog, Leu-1 B. Immunol Rev. 1986 Oct;93:53–79. doi: 10.1111/j.1600-065x.1986.tb01502.x. [DOI] [PubMed] [Google Scholar]
  15. Harper M. E., Marselle L. M., Gallo R. C., Wong-Staal F. Detection of lymphocytes expressing human T-lymphotropic virus type III in lymph nodes and peripheral blood from infected individuals by in situ hybridization. Proc Natl Acad Sci U S A. 1986 Feb;83(3):772–776. doi: 10.1073/pnas.83.3.772. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hartman A. B., Rudikoff S. VH genes encoding the immune response to beta-(1,6)-galactan: somatic mutation in IgM molecules. EMBO J. 1984 Dec 1;3(12):3023–3030. doi: 10.1002/j.1460-2075.1984.tb02249.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hengartner H., Meo T., Müller E. Assignment of genes for immunoglobulin kappa and heavy chains to chromosomes 6 and 12 in mouse. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4494–4498. doi: 10.1073/pnas.75.9.4494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Herzenberg L. A., Stall A. M., Lalor P. A., Sidman C., Moore W. A., Parks D. R., Herzenberg L. A. The Ly-1 B cell lineage. Immunol Rev. 1986 Oct;93:81–102. doi: 10.1111/j.1600-065x.1986.tb01503.x. [DOI] [PubMed] [Google Scholar]
  19. Hofman F. M., Billing R. J., Parker J. W., Taylor C. R. Cytoplasmic as opposed to surface Ia antigens expressed on human peripheral blood lymphocytes and monocytes. Clin Exp Immunol. 1982 Aug;49(2):355–363. [PMC free article] [PubMed] [Google Scholar]
  20. Honjo T. Immunoglobulin genes. Annu Rev Immunol. 1983;1:499–528. doi: 10.1146/annurev.iy.01.040183.002435. [DOI] [PubMed] [Google Scholar]
  21. Jeong H. D., Komisar J. L., Kraig E., Teale J. M. Strain-dependent expression of VH gene families. J Immunol. 1988 Apr 1;140(7):2436–2441. [PubMed] [Google Scholar]
  22. Klinman N. R., Press J. L. Expression of specific clones during B cell development. Fed Proc. 1975 Jan;34(1):47–50. [PubMed] [Google Scholar]
  23. Klinman N. R., Press J. L. The characterization fo the B-cell repertoire specific for the 2,4-dinitrophenyl and 2,4,6-trinitrophenyl determinants in neonatal BALB/c mice. J Exp Med. 1975 May 1;141(5):1133–1146. doi: 10.1084/jem.141.5.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lawler A. M., Lin P. S., Gearhart P. J. Adult B-cell repertoire is biased toward two heavy-chain variable-region genes that rearrange frequently in fetal pre-B cells. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2454–2458. doi: 10.1073/pnas.84.8.2454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Livant D., Blatt C., Hood L. One heavy chain variable region gene segment subfamily in the BALB/c mouse contains 500-1000 or more members. Cell. 1986 Nov 7;47(3):461–470. doi: 10.1016/0092-8674(86)90603-3. [DOI] [PubMed] [Google Scholar]
  26. Mage M. G., McHugh L. L., Rothstein T. L. Mouse lymphocytes with and without surface immunoglobulin: preparative scale separation in polystyrene tissue culture dishes coated with specifically purified anti-immunoglobulin. J Immunol Methods. 1977;15(1):47–56. doi: 10.1016/0022-1759(77)90016-3. [DOI] [PubMed] [Google Scholar]
  27. Malynn B. A., Berman J. E., Yancopoulos G. D., Bona C. A., Alt F. W. Expression of the immunoglobulin heavy-chain variable gene repertoire. Curr Top Microbiol Immunol. 1987;135:75–94. doi: 10.1007/978-3-642-71851-9_6. [DOI] [PubMed] [Google Scholar]
  28. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Myers C. D., Sanders V. M., Vitetta E. S. Isolation of antigen-binding virgin and memory B cells. J Immunol Methods. 1986 Aug 21;92(1):45–57. doi: 10.1016/0022-1759(86)90502-8. [DOI] [PubMed] [Google Scholar]
  30. Pardoll D. M., Fowlkes B. J., Lechler R. I., Germain R. N., Schwartz R. H. Early genetic events in T cell development analyzed by in situ hybridization. J Exp Med. 1987 Jun 1;165(6):1624–1638. doi: 10.1084/jem.165.6.1624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Perlmutter R. M., Kearney J. F., Chang S. P., Hood L. E. Developmentally controlled expression of immunoglobulin VH genes. Science. 1985 Mar 29;227(4694):1597–1601. doi: 10.1126/science.3975629. [DOI] [PubMed] [Google Scholar]
  32. Rathbun G. A., Capra J. D., Tucker P. W. Organization of the murine immunoglobulin VH complex in the inbred strains. EMBO J. 1987 Oct;6(10):2931–2937. doi: 10.1002/j.1460-2075.1987.tb02597.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Reth M. G., Jackson S., Alt F. W. VHDJH formation and DJH replacement during pre-B differentiation: non-random usage of gene segments. EMBO J. 1986 Sep;5(9):2131–2138. doi: 10.1002/j.1460-2075.1986.tb04476.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Riblet R., Brodeur P. H. The Igh-V gene repertoire of the mouse. Mt Sinai J Med. 1986 Mar;53(3):170–174. [PubMed] [Google Scholar]
  35. Riley S. C., Connors S. J., Klinman N. R., Ogata R. T. Preferential expression of variable region heavy chain gene segments by predominant 2,4-dinitrophenyl-specific BALB/c neonatal antibody clonotypes. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2589–2593. doi: 10.1073/pnas.83.8.2589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. SILVERSTEIN A. M., UHR J. W., KRANER K. L., LUKES R. J. Fetal response to antigenic stimulus. II. Antibody production by the fetal lamb. J Exp Med. 1963 May 1;117:799–812. doi: 10.1084/jem.117.5.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Schroeder H. W., Jr, Hillson J. L., Perlmutter R. M. Early restriction of the human antibody repertoire. Science. 1987 Nov 6;238(4828):791–793. doi: 10.1126/science.3118465. [DOI] [PubMed] [Google Scholar]
  38. Schulze D. H., Kelsoe G. Genotypic analysis of B cell colonies by in situ hybridization. Stoichiometric expression of three VH families in adult C57BL/6 and BALB/c mice. J Exp Med. 1987 Jul 1;166(1):163–172. doi: 10.1084/jem.166.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sherwin W. K., Rowlands D. T., Jr Determinants of the hierarchy of humoral immune responsiveness during ontogeny. J Immunol. 1975 Dec;115(6):1549–1554. [PubMed] [Google Scholar]
  40. Sigal N. H., Pickard A. R., Metcalf E. S., Gearhart P. J., Klinman N. R. Expression of phosphorylcholine-specific B cells during murine development. J Exp Med. 1977 Oct 1;146(4):933–948. doi: 10.1084/jem.146.4.933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Snow E. C., Vitetta E. S., Uhr J. W. Activation of antigen-enriched b cells. I. Purification and response to thymus-independent antigens. J Immunol. 1983 Feb;130(2):607–613. [PubMed] [Google Scholar]
  42. Sugiyama H., Maeda T., Tani Y., Miyake S., Oka Y., Komori T., Ogawa H., Soma T., Minami Y., Sakato N. Selective use of the VHQ52 family in functional VH to DJH rearrangements in a B precursor cell line. J Exp Med. 1987 Aug 1;166(2):607–612. doi: 10.1084/jem.166.2.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Teale J. M. B cell immune repertoire diversifies in a predictable temporal order in vitro. J Immunol. 1985 Aug;135(2):954–958. [PubMed] [Google Scholar]
  44. Teale J. M., Kearney J. F. Clonotypic analysis of the fetal B cell repertoire: evidence for an early and predominant expression of idiotypes associated with the VH 36-60 family. J Mol Cell Immunol. 1986;2(5):283–292. [PubMed] [Google Scholar]
  45. Teale J. M., Layton J. E., Nossal G. J. In vitro model for natural tolerance to self-antigens. Inhibition of the development of surface-immunoglobulin-negative lymphocytes into T-dependent responsive B cells by antigen. J Exp Med. 1979 Aug 1;150(2):205–217. doi: 10.1084/jem.150.2.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Teale J. M., Mandel T. E. Ontogenic development of B-lymphocyte function and tolerance susceptibility in vivo and in an in vitro organ culture system. J Exp Med. 1980 Feb 1;151(2):429–445. doi: 10.1084/jem.151.2.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983 Apr 14;302(5909):575–581. doi: 10.1038/302575a0. [DOI] [PubMed] [Google Scholar]
  48. Vakil M., Kearney J. F. Functional characterization of monoclonal auto-anti-idiotype antibodies isolated from the early B cell repertoire of BALB/c mice. Eur J Immunol. 1986 Sep;16(9):1151–1158. doi: 10.1002/eji.1830160920. [DOI] [PubMed] [Google Scholar]
  49. Weaver D., Reis M. H., Albanese C., Costantini F., Baltimore D., Imanishi-Kari T. Altered repertoire of endogenous immunoglobulin gene expression in transgenic mice containing a rearranged mu heavy chain gene. Cell. 1986 Apr 25;45(2):247–259. doi: 10.1016/0092-8674(86)90389-2. [DOI] [PubMed] [Google Scholar]
  50. Winter E., Radbruch A., Krawinkel U. Members of novel VH gene families are found in VDJ regions of polyclonally activated B-lymphocytes. EMBO J. 1985 Nov;4(11):2861–2867. doi: 10.1002/j.1460-2075.1985.tb04015.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wu G. E., Paige C. J. VH gene family utilization in colonies derived from B and pre-B cells detected by the RNA colony blot assay. EMBO J. 1986 Dec 20;5(13):3475–3481. doi: 10.1002/j.1460-2075.1986.tb04672.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Yancopoulos G. D., Desiderio S. V., Paskind M., Kearney J. F., Baltimore D., Alt F. W. Preferential utilization of the most JH-proximal VH gene segments in pre-B-cell lines. Nature. 1984 Oct 25;311(5988):727–733. doi: 10.1038/311727a0. [DOI] [PubMed] [Google Scholar]
  53. Zeldis J. B., Riblet R., Konigsberg W. H., Richards F. F., Rosenstein R. W. The location and expression of idiotypic determinants in the immunoglobulin variable region--III. Expression of the protein 315 and 460 idiotypic determinants in mouse anti-Dnp antibodies. Mol Immunol. 1979 Sep;16(9):657–664. doi: 10.1016/0161-5890(79)90005-1. [DOI] [PubMed] [Google Scholar]

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