Induction of several new antigens, including a T cell-associated antigen, following conversion of the EBV-negative B cell line RAMOS to EHRB-RAMOS by Epstein-Barr virus

K. S. Rosenthal, H. Shuman, J. L. Strominger

Research output: Contribution to journalArticle

Abstract

Heteroantisera prepared against EHRB-RAMOS cells and then absorbed extensively with the syngeneic EBV-negative cell line RAMOS recognized a membrane antigen present on EHRB-RAMOS but not on RAMOS cells. The binding of the sera, anti-EHRB, was measured by several immunologic techniques that detect cell surface molecules, including cytotoxicity, immunofluorescence, and the 125I-staph protein A assay. Triton X-100 extracts of plasma membranes prepared from EHRB-RAMOS, as measured by the staph protein A assay, whereas extracts of RAMOS membranes did not. The anti-EHRB sera also recognized the EHRA-RAMOS cell line, but binding of anti-EHRB to other EBV-converted RAMOS lines - African Burkitt lymphoma lines, newly established and well established lymphoblastoid cell lines, and EBV producer cell lines - was undetectable. The anti-EHRB sera recognized 4 T cell lines established from T cell leukemias, including CEM, HSB, YT4E, and MOLT 4. In addition, an anti-CEM serum, prepared in this laboratory, recognized the EHRB-RAMOS cells. Absorption of the serum with peripheral blood lymphocytes removed the activity of anti-EHRB to both the T cell lines and to EHRB-RAMOS, as detected by immunofluorescence using the fluorescence-activated cell sorter. The EHRB-RAMOS cells still retained B cell characteristics, since they expressed p29, 34, and antigen found on normal peripheral B cells, and did not rosette with sheep red blood cells. The anti-EHRB sera and anti-CEM sera were further characterized by radioimmunoprecipitation of 35S-methionine-labeled proteins of EHRB-RAMOS, RAMOS, YT4E, CEM, and HSB. Anti-EHRB recognized polypeptides of 16, 32, and 35 x 103 daltons on EHRB-RAMOS and 85 x 103 daltons on EHRB-RAMOS, CEM, HSB, and YT4E. Anti-CEM recognized polypeptides of 95 to 130 x 103 daltons expressed on the T cells and EHRB-RAMOS. These polypeptides were absent from or expressed to a much less extent on RAMOS. Thus conversion of RAMOS by EBV to the EHRB-RAMOS B cell line was associated with the appearance of several different antigens, some of which were also present on 4 T cell lines and on peripheral blood lymphocytes (as evidenced by absorption).

Original languageEnglish (US)
Pages (from-to)746-754
Number of pages9
JournalJournal of Immunology
Volume127
Issue number2
StatePublished - Jan 1 1981
Externally publishedYes

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Human Herpesvirus 4
B-Lymphocytes
T-Lymphocytes
Antigens
Cell Line
Serum
Burkitt Lymphoma
Staphylococcal Protein A
Peptides
Fluorescent Antibody Technique
Lymphocytes
Immunologic Techniques
T-Cell Leukemia
Membranes
Octoxynol
Methionine
Sheep
Erythrocytes
Fluorescence
Cell Membrane

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Induction of several new antigens, including a T cell-associated antigen, following conversion of the EBV-negative B cell line RAMOS to EHRB-RAMOS by Epstein-Barr virus. / Rosenthal, K. S.; Shuman, H.; Strominger, J. L.

In: Journal of Immunology, Vol. 127, No. 2, 01.01.1981, p. 746-754.

Research output: Contribution to journalArticle

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abstract = "Heteroantisera prepared against EHRB-RAMOS cells and then absorbed extensively with the syngeneic EBV-negative cell line RAMOS recognized a membrane antigen present on EHRB-RAMOS but not on RAMOS cells. The binding of the sera, anti-EHRB, was measured by several immunologic techniques that detect cell surface molecules, including cytotoxicity, immunofluorescence, and the 125I-staph protein A assay. Triton X-100 extracts of plasma membranes prepared from EHRB-RAMOS, as measured by the staph protein A assay, whereas extracts of RAMOS membranes did not. The anti-EHRB sera also recognized the EHRA-RAMOS cell line, but binding of anti-EHRB to other EBV-converted RAMOS lines - African Burkitt lymphoma lines, newly established and well established lymphoblastoid cell lines, and EBV producer cell lines - was undetectable. The anti-EHRB sera recognized 4 T cell lines established from T cell leukemias, including CEM, HSB, YT4E, and MOLT 4. In addition, an anti-CEM serum, prepared in this laboratory, recognized the EHRB-RAMOS cells. Absorption of the serum with peripheral blood lymphocytes removed the activity of anti-EHRB to both the T cell lines and to EHRB-RAMOS, as detected by immunofluorescence using the fluorescence-activated cell sorter. The EHRB-RAMOS cells still retained B cell characteristics, since they expressed p29, 34, and antigen found on normal peripheral B cells, and did not rosette with sheep red blood cells. The anti-EHRB sera and anti-CEM sera were further characterized by radioimmunoprecipitation of 35S-methionine-labeled proteins of EHRB-RAMOS, RAMOS, YT4E, CEM, and HSB. Anti-EHRB recognized polypeptides of 16, 32, and 35 x 103 daltons on EHRB-RAMOS and 85 x 103 daltons on EHRB-RAMOS, CEM, HSB, and YT4E. Anti-CEM recognized polypeptides of 95 to 130 x 103 daltons expressed on the T cells and EHRB-RAMOS. These polypeptides were absent from or expressed to a much less extent on RAMOS. Thus conversion of RAMOS by EBV to the EHRB-RAMOS B cell line was associated with the appearance of several different antigens, some of which were also present on 4 T cell lines and on peripheral blood lymphocytes (as evidenced by absorption).",
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T1 - Induction of several new antigens, including a T cell-associated antigen, following conversion of the EBV-negative B cell line RAMOS to EHRB-RAMOS by Epstein-Barr virus

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N2 - Heteroantisera prepared against EHRB-RAMOS cells and then absorbed extensively with the syngeneic EBV-negative cell line RAMOS recognized a membrane antigen present on EHRB-RAMOS but not on RAMOS cells. The binding of the sera, anti-EHRB, was measured by several immunologic techniques that detect cell surface molecules, including cytotoxicity, immunofluorescence, and the 125I-staph protein A assay. Triton X-100 extracts of plasma membranes prepared from EHRB-RAMOS, as measured by the staph protein A assay, whereas extracts of RAMOS membranes did not. The anti-EHRB sera also recognized the EHRA-RAMOS cell line, but binding of anti-EHRB to other EBV-converted RAMOS lines - African Burkitt lymphoma lines, newly established and well established lymphoblastoid cell lines, and EBV producer cell lines - was undetectable. The anti-EHRB sera recognized 4 T cell lines established from T cell leukemias, including CEM, HSB, YT4E, and MOLT 4. In addition, an anti-CEM serum, prepared in this laboratory, recognized the EHRB-RAMOS cells. Absorption of the serum with peripheral blood lymphocytes removed the activity of anti-EHRB to both the T cell lines and to EHRB-RAMOS, as detected by immunofluorescence using the fluorescence-activated cell sorter. The EHRB-RAMOS cells still retained B cell characteristics, since they expressed p29, 34, and antigen found on normal peripheral B cells, and did not rosette with sheep red blood cells. The anti-EHRB sera and anti-CEM sera were further characterized by radioimmunoprecipitation of 35S-methionine-labeled proteins of EHRB-RAMOS, RAMOS, YT4E, CEM, and HSB. Anti-EHRB recognized polypeptides of 16, 32, and 35 x 103 daltons on EHRB-RAMOS and 85 x 103 daltons on EHRB-RAMOS, CEM, HSB, and YT4E. Anti-CEM recognized polypeptides of 95 to 130 x 103 daltons expressed on the T cells and EHRB-RAMOS. These polypeptides were absent from or expressed to a much less extent on RAMOS. Thus conversion of RAMOS by EBV to the EHRB-RAMOS B cell line was associated with the appearance of several different antigens, some of which were also present on 4 T cell lines and on peripheral blood lymphocytes (as evidenced by absorption).

AB - Heteroantisera prepared against EHRB-RAMOS cells and then absorbed extensively with the syngeneic EBV-negative cell line RAMOS recognized a membrane antigen present on EHRB-RAMOS but not on RAMOS cells. The binding of the sera, anti-EHRB, was measured by several immunologic techniques that detect cell surface molecules, including cytotoxicity, immunofluorescence, and the 125I-staph protein A assay. Triton X-100 extracts of plasma membranes prepared from EHRB-RAMOS, as measured by the staph protein A assay, whereas extracts of RAMOS membranes did not. The anti-EHRB sera also recognized the EHRA-RAMOS cell line, but binding of anti-EHRB to other EBV-converted RAMOS lines - African Burkitt lymphoma lines, newly established and well established lymphoblastoid cell lines, and EBV producer cell lines - was undetectable. The anti-EHRB sera recognized 4 T cell lines established from T cell leukemias, including CEM, HSB, YT4E, and MOLT 4. In addition, an anti-CEM serum, prepared in this laboratory, recognized the EHRB-RAMOS cells. Absorption of the serum with peripheral blood lymphocytes removed the activity of anti-EHRB to both the T cell lines and to EHRB-RAMOS, as detected by immunofluorescence using the fluorescence-activated cell sorter. The EHRB-RAMOS cells still retained B cell characteristics, since they expressed p29, 34, and antigen found on normal peripheral B cells, and did not rosette with sheep red blood cells. The anti-EHRB sera and anti-CEM sera were further characterized by radioimmunoprecipitation of 35S-methionine-labeled proteins of EHRB-RAMOS, RAMOS, YT4E, CEM, and HSB. Anti-EHRB recognized polypeptides of 16, 32, and 35 x 103 daltons on EHRB-RAMOS and 85 x 103 daltons on EHRB-RAMOS, CEM, HSB, and YT4E. Anti-CEM recognized polypeptides of 95 to 130 x 103 daltons expressed on the T cells and EHRB-RAMOS. These polypeptides were absent from or expressed to a much less extent on RAMOS. Thus conversion of RAMOS by EBV to the EHRB-RAMOS B cell line was associated with the appearance of several different antigens, some of which were also present on 4 T cell lines and on peripheral blood lymphocytes (as evidenced by absorption).

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