Abstract
Accessible surface proteins of thymus-derived lymphocytes (T-cells) of normal CBA mice and bone-marrow-derived lymphocytes (B-cells) of congenitally athymic nu/nu mice were analysed. The surfaces of lymphocytes were radioiodinated by using the enzyme lactoperoxidase (EC 1.11.1.7), then solubilized either in acid–urea or in the non-ionic detergent Nonidet P-40. These lysates were then precipitated with antisera specific to either immunoglobulin or the θ-alloantigen in order to assess the presence of these surface markers. Comparable amounts of radioactivity in proteins specifically precipitable as immunoglobulin were obtained from T-lymphocytes and B-lymphocytes when the cells were disrupted by acid–urea. This immunoglobulin had mol. wt. approx. 180000 and was composed of light chains and μ-type heavy chains. When radioiodinated lymphocytes were solubilized with Nonidet P-40, 3–4% of radioiodinated high-molecular-weight protein of B-cells consisted of immunoglobulin, a result similar to that found with acid–urea extraction. However, with the detergent extraction, only 0.1% of T-cell surface protein was precipitable by anti-globulin reagents. The θ-alloantigen was isolated from CBA T-cells both by acid–urea and by detergent lysis. This protein possessed a mobility on polyacrylamide-gel electrophoresis in sodium dodecyl sulphate which was consistent with a mol. wt. of 60000. An identical component was isolated from the θ-positive thymoma WEHI 105. The θ-antigen was not isolated from B-cells by either of the extraction procedures used. These results provide further evidence that the surface membranes of normal T-cells and B-cells differ in physicochemical properties. In particular, various surface components possess differential solubilities in non-ionic or organic solvents. This observation provides an explanation for discrepant results that have appeared in the literature concerning the isolation of immunoglobulin from T-lymphocytes.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allan D., Crumpton M. J. Solubilization of pig lymphocyte plasma membrane and fractionation of some of the components. Biochem J. 1971 Aug;123(5):967–975. doi: 10.1042/bj1230967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Andersson J., Sjöberg O., Möller G. Mitogens as probes for immunocyte activation and cellular cooperation. Transplant Rev. 1972;11:131–177. doi: 10.1111/j.1600-065x.1972.tb00048.x. [DOI] [PubMed] [Google Scholar]
- Atwell J. L., Cone R. E., Marchalonis J. J. Isolation of theta antigen from the surface of thymus lymphocytes. Nat New Biol. 1973 Feb 21;241(112):251–252. doi: 10.1038/newbio241251a0. [DOI] [PubMed] [Google Scholar]
- Baur S., Vitetta E. S., Sherr C. J., Schenkein I., Uhr J. W. Isolation of heavy and light chains of immunoglobulin from the surfaces of lymphoid cells. J Immunol. 1971 Apr;106(4):1133–1135. [PubMed] [Google Scholar]
- Cotman C. W., Banker G., Levy W., Taylor D. An ultrastructural and chemical analysis of the effect of triton X-100 on synaptic plasma membranes. Biochim Biophys Acta. 1971 Dec 3;249(2):406–418. doi: 10.1016/0005-2736(71)90119-2. [DOI] [PubMed] [Google Scholar]
- Crone M., Koch C., Simonsen M. The elusive T cell receptor. Transplant Rev. 1972;10:36–56. doi: 10.1111/j.1600-065x.1972.tb01538.x. [DOI] [PubMed] [Google Scholar]
- De Sousa M. A., Parrott D. M., Pantelouris E. M. The lymphoid tissues in mice with congenital aplasia of the thymus. Clin Exp Immunol. 1969 Jun;4(6):637–644. [PMC free article] [PubMed] [Google Scholar]
- Flanagan S. P. 'Nude', a new hairless gene with pleiotropic effects in the mouse. Genet Res. 1966 Dec;8(3):295–309. doi: 10.1017/s0016672300010168. [DOI] [PubMed] [Google Scholar]
- Greaves M., Janossy G. Elicitation of selective T and B lymphocyte responses by cell surface binding ligands. Transplant Rev. 1972;11:87–130. doi: 10.1111/j.1600-065x.1972.tb00047.x. [DOI] [PubMed] [Google Scholar]
- Harris A. W., Bankhurst A. D., Mason S., Warner N. L. Differentiated functions expressed by cultured mouse lymphoma cells. II. Theta antigen, surface immunoglobulin and a receptor for antibody on cells of a thymoma cell line. J Immunol. 1973 Feb;110(2):431–438. [PubMed] [Google Scholar]
- Jones G. Radiolabeling of cell surface alloantigens by the lactoperoxidase method. Transplantation. 1972 Nov;14(5):655–657. doi: 10.1097/00007890-197211000-00021. [DOI] [PubMed] [Google Scholar]
- 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]
- Mandel T. E. Intramembraneous marker in T lymphocytes. Nat New Biol. 1972 Sep 27;239(91):112–114. doi: 10.1038/newbio239112a0. [DOI] [PubMed] [Google Scholar]
- Marchalonis J. J., Cone R. E., Atwell J. L. Isolation and partial characterization of lymphocyte surface immunoglobulins. J Exp Med. 1972 Apr 1;135(4):956–971. doi: 10.1084/jem.135.4.956. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marchalonis J. J., Cone R. E., Santer V. Enzymic iodination. A probe for accessible surface proteins of normal and neoplastic lymphocytes. Biochem J. 1971 Oct;124(5):921–927. doi: 10.1042/bj1240921. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moroz C., Hahn Y. Cell-surface immunoglobulin human thymus cells and its biosynthesis in vitro. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3716–3720. doi: 10.1073/pnas.70.12.3716. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Phillips D. R., Morrison M. Changes in accessibility of plasma membrane protein as the result of tryptic hydrolysis. Nat New Biol. 1973 Apr 18;242(120):213–215. doi: 10.1038/newbio242213a0. [DOI] [PubMed] [Google Scholar]
- Polliack A., Lampen N., Clarkson B. D., De Harven E., Bentwich Z., Siegal F. P., Kunkel H. G. Identification of human B and T lymphocytes by scanning electron microscopy. J Exp Med. 1973 Sep 1;138(3):607–624. doi: 10.1084/jem.138.3.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raff M. C. Surface antigenic markers for distinguishing T and B lymphocytes in mice. Transplant Rev. 1971;6:52–80. doi: 10.1111/j.1600-065x.1971.tb00459.x. [DOI] [PubMed] [Google Scholar]
- Santer V., Cone R. E., Marchalonis J. J. The glycoprotein surface coat on different classes of murine lymphocytes. Exp Cell Res. 1973 Jun;79(2):404–416. doi: 10.1016/0014-4827(73)90460-6. [DOI] [PubMed] [Google Scholar]
- Siekevitz P. Biological membranes: the dynamics of their organization. Annu Rev Physiol. 1972;34:117–140. doi: 10.1146/annurev.ph.34.030172.001001. [DOI] [PubMed] [Google Scholar]
- Vitetta E. S., Bianco C., Nussenzweig V., Uhr J. W. Cell surface immunoglobulin. IV. Distribution among thymocytes, bone mrrow cells, and their derived populations. J Exp Med. 1972 Jul 1;136(1):81–93. doi: 10.1084/jem.136.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vitetta E. S., Bianco C., Nussenzweig V., Uhr J. W. Cell surface immunoglobulin. IV. Distribution among thymocytes, bone mrrow cells, and their derived populations. J Exp Med. 1972 Jul 1;136(1):81–93. doi: 10.1084/jem.136.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vitetta E. S., Boyse E. A., Uhr J. W. Isolation and characterization of a molecular complex containing Thy-1 antigen from the surface of murine thymocytes and T cells. Eur J Immunol. 1973 Jul;3(7):446–453. doi: 10.1002/eji.1830030714. [DOI] [PubMed] [Google Scholar]