Abstract
15 SM/J mouse hybridoma antibodies that show antithymocyte autoantibody (ATA) activity by immunofluorescence staining were studied. Half of these antibodies react with determinants whose expression is associated with Thy-1, as shown by blocking experiments with anti-Thy-1 and loss of reactivity with Thy-1- mutant cell lines. The Thy-1 dependence of three of these ATA is further confirmed by their reexpression on a Thy- 1 gene transfectant. However, the remaining antibodies exhibited binding that showed little or no dependence on Thy-1. Furthermore, we find that most ATA derives from the Ly-1 B subpopulation, as demonstrated by lipopolysaccharide-induced ATA secretion in vitro and by comparison of ATA hybridoma frequencies. VH region gene sequence data of 14 monoclonal ATA from Ly-1 B cell-derived hybridomas reveal the utilization of nine VH genes belonging to four different VH families (J558, 3609, Q52, and Vgam3.8). While we find that two of these hybridomas arose from a clonal expansion, we also find four examples of a 3609 family VH gene utilized in clonally independent lines showing similar specificity. Yet another example of identical VH gene usage by clonally unrelated cells is found in two J558 ATA of a distinct fine specificity. These data suggest that the enrichment of ATA B cells in the Ly-1 B subset is primarily due to repeated independent recruitment of B cells by antigen resulting in the expression of a restricted set of VH genes.
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- Auchincloss H., Jr, Ozato K., Sachs D. H. A monoclonal antibody detecting unusual Thy-1 determinants. J Immunol. 1982 Apr;128(4):1584–1589. [PubMed] [Google Scholar]
- BOYDEN S. AUTOIMMUNITY AND INFLAMMATION. Nature. 1964 Jan 11;201:200–201. doi: 10.1038/201200a0. [DOI] [PubMed] [Google Scholar]
- Barclay A. N., Letarte-Muirhead M., Williams A. F., Faulkes R. A. Chemical characterisation of the Thy-1 glycoproteins from the membranes of rat thymocytes and brain. Nature. 1976 Oct 14;263(5578):563–567. doi: 10.1038/263563a0. [DOI] [PubMed] [Google Scholar]
- Bray K. R., Gershwin M. E., Ahmed A., Castles J. J. Tissue localization and biochemical characteristics of a new thymic antigen recognized by a monoclonal thymocytotoxic autoantibody from New Zealand black mice. J Immunol. 1985 Jun;134(6):4001–4008. [PubMed] [Google Scholar]
- Carmack C. E., Shinton S. A., Hayakawa K., Hardy R. R. Rearrangement and selection of VH11 in the Ly-1 B cell lineage. J Exp Med. 1990 Jul 1;172(1):371–374. doi: 10.1084/jem.172.1.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chapman A., Fujimoto K., Kornfeld S. The primary glycosylation defect in class E Thy-1-negative mutant mouse lymphoma cells is an inability to synthesize dolichol-P-mannose. J Biol Chem. 1980 May 25;255(10):4441–4446. [PubMed] [Google Scholar]
- Conzelmann A., Spiazzi A., Hyman R., Bron C. Anchoring of membrane proteins via phosphatidylinositol is deficient in two classes of Thy-1 negative mutant lymphoma cells. EMBO J. 1986 Dec 1;5(12):3291–3296. doi: 10.1002/j.1460-2075.1986.tb04642.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cunningham A. J. Large numbers of cells in normal mice produce antibody components of isologous erythrocytes. Nature. 1974 Dec 20;252(5485):749–751. doi: 10.1038/252749a0. [DOI] [PubMed] [Google Scholar]
- Eisenberg R. A., Theofilopoulos A. N., Andrews B. S., Peters C. J., Thor L., Dixon F. J. Natural thymocytotoxic autoantibodies in autoimmune and normal mice. J Immunol. 1979 Jun;122(6):2272–2278. [PubMed] [Google Scholar]
- Evans G. A., Hyman R., Lewis K. A mutant lymphoma cell line with a defective Thy-1 glycoprotein gene. Immunogenetics. 1987;25(1):28–34. doi: 10.1007/BF00768830. [DOI] [PubMed] [Google Scholar]
- Hardy R. R., Carmack C. E., Shinton S. A., Riblet R. J., Hayakawa K. A single VH gene is utilized predominantly in anti-BrMRBC hybridomas derived from purified Ly-1 B cells. Definition of the VH11 family. J Immunol. 1989 May 15;142(10):3643–3651. [PubMed] [Google Scholar]
- Haughton G., Arnold L. W., Bishop G. A., Mercolino T. J. The CH series of murine B cell lymphomas: neoplastic analogues of Ly-1+ normal B cells. Immunol Rev. 1986 Oct;93:35–51. doi: 10.1111/j.1600-065x.1986.tb01501.x. [DOI] [PubMed] [Google Scholar]
- Hayakawa K., Hardy R. R., Herzenberg L. A., Herzenberg L. A. Progenitors for Ly-1 B cells are distinct from progenitors for other B cells. J Exp Med. 1985 Jun 1;161(6):1554–1568. doi: 10.1084/jem.161.6.1554. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayakawa K., Hardy R. R., Honda M., Herzenberg L. A., Steinberg A. D., Herzenberg L. A. Ly-1 B cells: functionally distinct lymphocytes that secrete IgM autoantibodies. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2494–2498. doi: 10.1073/pnas.81.8.2494. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayakawa K., Hardy R. R. Murine CD4+ T cell subsets defined. J Exp Med. 1988 Nov 1;168(5):1825–1838. doi: 10.1084/jem.168.5.1825. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayakawa K., Hardy R. R. Normal, autoimmune, and malignant CD5+ B cells: the Ly-1 B lineage? Annu Rev Immunol. 1988;6:197–218. doi: 10.1146/annurev.iy.06.040188.001213. [DOI] [PubMed] [Google Scholar]
- Hayakawa K., Hardy R. R., Parks D. R., Herzenberg L. A. The "Ly-1 B" cell subpopulation in normal immunodefective, and autoimmune mice. J Exp Med. 1983 Jan 1;157(1):202–218. doi: 10.1084/jem.157.1.202. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayakawa K., Hardy R. R. Phenotypic and functional alteration of CD4+ T cells after antigen stimulation. Resolution of two populations of memory T cells that both secrete interleukin 4. J Exp Med. 1989 Jun 1;169(6):2245–2250. doi: 10.1084/jem.169.6.2245. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hyman R., Stallings V. Complementation patterns of Thy-1 variants and evidence that antigen loss variants "pre-exist" in the parental population. J Natl Cancer Inst. 1974 Feb;52(2):429–436. doi: 10.1093/jnci/52.2.429. [DOI] [PubMed] [Google Scholar]
- Imai Y., Nakano T., Sawada J. I., Osawa T. Specificity of natural thymocytotoxic autoantibody developed in New Zealand black mice. J Immunol. 1980 Apr;124(4):1556–1561. [PubMed] [Google Scholar]
- Isobe K., Fortunato A., Giguere V., Grosveld F., Mitchison N. A. Anti-Thy-1 antibody responses evoked by Thy-1 antigen expressed in transfected mouse mastocytoma cells and rat fibroblast. Immunology. 1985 Nov;56(3):505–512. [PMC free article] [PubMed] [Google Scholar]
- Kipps T. J. The CD5 B cell. Adv Immunol. 1989;47:117–185. doi: 10.1016/s0065-2776(08)60663-x. [DOI] [PubMed] [Google Scholar]
- Lefrancois L., Puddington L., Machamer C. E., Bevan M. J. Acquisition of cytotoxic T lymphocyte-specific carbohydrate differentiation antigens. J Exp Med. 1985 Oct 1;162(4):1275–1293. doi: 10.1084/jem.162.4.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marcu K. B., Banerji J., Penncavage N. A., Lang R., Arnheim N. 5' flanking region of immunoglobulin heavy chain constant region genes displays length heterogeneity in germlines of inbred mouse strains. Cell. 1980 Nov;22(1 Pt 1):187–196. doi: 10.1016/0092-8674(80)90167-1. [DOI] [PubMed] [Google Scholar]
- Martin W. J., Martin S. E. Naturally occurring cytotoxic anti-tumour antibodies in sera of congenitally athymic (nude) mice. Nature. 1974 Jun 7;249(457):564–565. doi: 10.1038/249564a0. [DOI] [PubMed] [Google Scholar]
- Martin W. J., Martin S. E. Thymus reactive IgM autoantibodies in normal mouse sera. Nature. 1975 Apr 24;254(5502):716–718. doi: 10.1038/254716a0. [DOI] [PubMed] [Google Scholar]
- Mulligan R. C., Berg P. Expression of a bacterial gene in mammalian cells. Science. 1980 Sep 19;209(4463):1422–1427. doi: 10.1126/science.6251549. [DOI] [PubMed] [Google Scholar]
- NORINS L. C., HOLMES M. C. ANTINUCLEAR FACTOR IN MICE. J Immunol. 1964 Jul;93:148–154. [PubMed] [Google Scholar]
- Parker L. M., Chused T. M., Steinberg A. D. Immunofluorescence studies on thymocytotoxic antibody from New Zealand Black mice. J Immunol. 1974 Jan;112(1):285–292. [PubMed] [Google Scholar]
- Pennell C. A., Arnold L. W., Haughton G., Clarke S. H. Restricted Ig variable region gene expression among Ly-1+ B cell lymphomas. J Immunol. 1988 Oct 15;141(8):2788–2796. [PubMed] [Google Scholar]
- Rabinovitch P. S., Torres R. M., Engel D. Simultaneous cell cycle analysis and two-color surface immunofluorescence using 7-amino-actinomycin D and single laser excitation: applications to study of cell activation and the cell cycle of murine Ly-1 B cells. J Immunol. 1986 Apr 15;136(8):2769–2775. [PubMed] [Google Scholar]
- Schlesinger M. Spontaneous occurrence of autoantibodies cytotoxic to thymus cells in the sera of mice of the 129 strain. Nature. 1965 Jul 24;207(995):429–430. doi: 10.1038/207429b0. [DOI] [PubMed] [Google Scholar]
- Seldin M. F., Conroy J., Steinberg A. D., D'Hoosteleare L. A., Raveche E. S. Clonal expansion of abnormal B cells in old NZB mice. J Exp Med. 1987 Nov 1;166(5):1585–1590. doi: 10.1084/jem.166.5.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shirai T., Hayakawa K., Okumura K., Tada T. Differential cytotoxic effect of natural thymocytotoxic autoantibody of NZB mice on functional subsets of T cells. J Immunol. 1978 Jun;120(6):1924–1929. [PubMed] [Google Scholar]
- Shirai T., Mellors R. C. Natural thymocytotoxic autoantibody and reactive antigen in New Zealand black and other mice. Proc Natl Acad Sci U S A. 1971 Jul;68(7):1412–1415. doi: 10.1073/pnas.68.7.1412. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Surh C. D., Gershwin M. E., Ahmed A. A peripheral and central T cell antigen recognized by a monoclonal thymocytotoxic autoantibody from New Zealand black mice. J Immunol. 1987 Mar 1;138(5):1421–1428. [PubMed] [Google Scholar]
- Tarlinton D., Stall A. M., Herzenberg L. A. Repetitive usage of immunoglobulin VH and D gene segments in CD5+ Ly-1 B clones of (NZB x NZW)F1 mice. EMBO J. 1988 Dec 1;7(12):3705–3710. doi: 10.1002/j.1460-2075.1988.tb03253.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas D. B., Giguere V., Graham C. M., Oliveira O. L. Autoimmunity to Thy-1. Eur J Immunol. 1986 Jan;16(1):40–47. doi: 10.1002/eji.1830160109. [DOI] [PubMed] [Google Scholar]
- Trowbridge I. S., Hyman R., Mazauskas C. The synthesis and properties of T25 blycoprotein in Thy-1-negative mutant lymphoma cells. Cell. 1978 May;14(1):21–32. doi: 10.1016/0092-8674(78)90297-0. [DOI] [PubMed] [Google Scholar]
- Zhang H., Scholl R., Browse J., Somerville C. Double stranded DNA sequencing as a choice for DNA sequencing. Nucleic Acids Res. 1988 Feb 11;16(3):1220–1220. doi: 10.1093/nar/16.3.1220. [DOI] [PMC free article] [PubMed] [Google Scholar]