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. 1981 Dec;78(12):7697–7701. doi: 10.1073/pnas.78.12.7697

Detection of T cells that secrete molecules which share determinants with antigen-specific T-cell factors.

W L Cleveland, I Wood, R E Cone, G M Iverson, R W Rosenstein, R K Gershon, B F Erlanger
PMCID: PMC349337  PMID: 6174978

Abstract

A single-cell secretion assay was used to detect cells that secrete products which react with an antiserum that binds T cell antigen-binding polypeptides. The antiserum (R11), which was produced by immunization of rabbits with a murine trinitrophenyl-specific suppressor factor, reacts with T cells and their products and with a suppressor T-cell clone but not with B cells or their products. The secretory cells this antiserum detected were found to be unevenly distributed among various organs (spleen, lymph node, and thymus) and, to different degrees, in spleens of various strains of mice. In unimmunized CBA/J mice, approximately 1-3% of spleen cells secreted macromolecules precipitable by R11. The majority of the secretory cells could be removed by panning with a mixture of Ly 1 and Ly 2 antibodies but not with either antibody alone. This is consistent with the cells having low surface antigen densities as a result of being either "pre-T" cells or mature secretory cells analogous to B-lineage plasma cells. In agreement with the latter possibility was our finding that the secretory activity of cells detected with antisuppressor factor was comparable to that of Ig-secretory cells as detected with an anti-Ig antiserum. However, higher numbers of R11+ secretory cells were seen in the immunoglobulin-negative fraction of spleen cells from nude mice, which could be interpreted to favor the first possibility. In either case this study shows that the single-cell assay technique is well suited for the detection and characterization of molecules released by immunoregulatory T cells.

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Selected References

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  1. Broker T. R., Angerer L. M., Yen P. H., Hershey N. D., Davidson N. Electron microscopic visualization of tRNA genes with ferritin-avidin: biotin labels. Nucleic Acids Res. 1978 Feb;5(2):363–384. doi: 10.1093/nar/5.2.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cambiaso C. L., Goffinet A., Vaerman J. P., Heremans J. F. Glutaraldehyde-activated aminohexyl- derivative of Sepharose 4B as a new verstile immunoabsorbent. Immunochemistry. 1975 Apr;12(4):273–278. doi: 10.1016/0019-2791(75)90175-5. [DOI] [PubMed] [Google Scholar]
  3. Cantor H., Boyse E. A. Lymphocytes as models for the study of mammalian cellular differentiation. Immunol Rev. 1977 Jan;33:105–124. doi: 10.1111/j.1600-065x.1977.tb00364.x. [DOI] [PubMed] [Google Scholar]
  4. Cantor H., Gershon R. K. Immunological circuits: cellular composition. Fed Proc. 1979 Jun;38(7):2058–2064. [PubMed] [Google Scholar]
  5. Cleveland W. L., Erlanger B. F. A general method for studying the secretion of macromolecules by single cells. I. Detection of immunoglobulin-secreting cells. Cell Immunol. 1978 Apr;37(1):229–242. doi: 10.1016/0008-8749(78)90190-9. [DOI] [PubMed] [Google Scholar]
  6. Cleveland W. L., Wood I., Jacobs I. N., Erlanger B. F. A general method for studying the secretion of macromolecules by single cells. II. A simplified procedure with enhanced sensitivity. Cell Immunol. 1981 Sep 1;63(1):154–163. doi: 10.1016/0008-8749(81)90036-8. [DOI] [PubMed] [Google Scholar]
  7. Cone R. E., Rosenstein R. W., Murray J. H., Iverson G. M., Ptak W., Gershon R. K. Characterization of T-cell surface proteins bound by heterologous antisera to antigen-specific T-cell products. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6411–6415. doi: 10.1073/pnas.78.10.6411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fresno M., Nabel G., McVay-Boudreau L., Furthmayer H., Cantor H. Antigen-specific T lymphocyte clones. I. Characterization of a T lymphocyte clone expressing antigen-specific suppressive activity. J Exp Med. 1981 May 1;153(5):1246–1259. doi: 10.1084/jem.153.5.1246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Grimm E., Bonavida B. Mechanism of cell-mediated cytotoxicity at the single cell level. I. Estimation of cytotoxic T lymphocyte frequency and relative lytic efficiency. J Immunol. 1979 Dec;123(6):2861–2869. [PubMed] [Google Scholar]
  10. Guesdon J. L., Ternynck T., Avrameas S. The use of avidin-biotin interaction in immunoenzymatic techniques. J Histochem Cytochem. 1979 Aug;27(8):1131–1139. doi: 10.1177/27.8.90074. [DOI] [PubMed] [Google Scholar]
  11. Ledbetter J. A., Herzenberg L. A. Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens. Immunol Rev. 1979;47:63–90. doi: 10.1111/j.1600-065x.1979.tb00289.x. [DOI] [PubMed] [Google Scholar]
  12. Ledbetter J. A., Rouse R. V., Micklem H. S., Herzenberg L. A. T cell subsets defined by expression of Lyt-1,2,3 and Thy-1 antigens. Two-parameter immunofluorescence and cytotoxicity analysis with monoclonal antibodies modifies current views. J Exp Med. 1980 Aug 1;152(2):280–295. doi: 10.1084/jem.152.2.280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Loor F., Roelants G. E. Immunofluorescence studies of a possible prethymic T-cell differentiation in congenitally athymic (nude) mice. Ann N Y Acad Sci. 1975 Jun 30;254:226–241. doi: 10.1111/j.1749-6632.1975.tb29173.x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Primi D., Lewis G. K., Goodman J. W. A hemolytic plaque assay for activated murine T cells. J Exp Med. 1979 Oct 1;150(4):987–1000. doi: 10.1084/jem.150.4.987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ptak W., Zembala M., Gershon R. K. Intermediary role of macrophages in the passage of suppressor signals between T-cell subsets. J Exp Med. 1978 Aug 1;148(2):424–434. doi: 10.1084/jem.148.2.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Shortman K., Williams N., Adams P. The separation of different cell classes from lymphoid organs. V. Simple procedures for the removal of cell debris. Damaged cells and erythroid cells from lymphoid cell suspensions. J Immunol Methods. 1972 May;1(3):273–287. doi: 10.1016/0022-1759(72)90005-1. [DOI] [PubMed] [Google Scholar]
  18. Steele E. J., Cunningham A. J. High proportion of Ig-producing cells making autoantibody in normal mice. Nature. 1978 Aug 3;274(5670):483–484. doi: 10.1038/274483a0. [DOI] [PubMed] [Google Scholar]
  19. Wysocki L. J., Sato V. L. "Panning" for lymphocytes: a method for cell selection. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2844–2848. doi: 10.1073/pnas.75.6.2844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. von Boehmer H., Shortman K. The separation of different cell classes from lymphoid organs. IX. A simple and rapid method for removal of damaged cells from lymphoid cell suspensions. J Immunol Methods. 1973 Apr;2(3):293–301. doi: 10.1016/0022-1759(73)90055-0. [DOI] [PubMed] [Google Scholar]

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