Characterization of antigens on human lymphocytes coded by messenger RNA translated in vitro

Robert Allen, Soldano Ferrone, James A. Hoch

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Translation and immunoprecipitation were used to identify messenger RNAs (mRNAs) coding for surface antigens expressed on human lymphoblastoid cells. The mRNAs were extracted from several human lymphoid cell lines as well as from fibroblastoid lines. These mRNAs were translated in vitro, and the translation products were reacted with xenoantisera raised against the antigens on human lymphoid cells. Products immunoprecipitated by these antisera were analysed by electrophoresis and fluorography. Four antisera immunoprecipitated a polypeptide with a mol. wt (MW) of approximately 32,000 (p32) from translations programmed with mRNA extracted from all the cell lines. Two antisera immunoprecipitated, in addition to p32. another polypeptide with a MW of approximately 25,000 (p25) only from translations programmed with RNA from lymphoid cell lines. p25 mRNA in the different lymphoid cell lines fell into three basic abundance classes as determined by in vitro translation and immunoprecipitation. Cells from two Burkitt's lymphomas (Raji and Daudi) did not express detectable p25 mRNA. Two T-lymphoblastoid lines (Molt-4 and 1301) contained five- to 10-fold less p25 mRNA than the B-lymphoid cell lines (Victor, RPMI-8866, RPMI-6410. RPM1-8226 and RPMI-1788). Both p32 and p25 were expressed on the cell surface inasmuch as lymphoblastoid cells adsorbed antibodies to both polypeptides. Human fibroblast, Raji or Daudi cells adsorbed anti-p32 antibodies from the antiserum but not anti-p25. Quantitative absorptions of the antiserum with T- or B-lymphoblastoid cells was used to determine the relative amounts of p32 and p25 expressed on the cell surface. B-Lymphoblastoid cells were found to express two- to five-fold more p25 on the cell surface than T-lymphoblastoid cells. p25 does not represent an immunoglobulin light-chain precursor inasmuch as a 1000-fold excess of unlabeled human Ig did not compete with p25 translated in vitro for binding by its respective antibody.

Original languageEnglish
Pages (from-to)1127-1138
Number of pages12
JournalMolecular Immunology
Volume19
Issue number9
DOIs
StatePublished - 1 Jan 1982

Fingerprint

Lymphocytes
Antigens
Messenger RNA
Immune Sera
Cell Line
Immunoprecipitation
Peptides
Photofluorography
In Vitro Techniques
Immunoglobulin Light Chains
Burkitt Lymphoma
Antibodies
Surface Antigens
Electrophoresis
Anti-Idiotypic Antibodies
Fibroblasts
RNA

Cite this

Allen, Robert ; Ferrone, Soldano ; Hoch, James A. / Characterization of antigens on human lymphocytes coded by messenger RNA translated in vitro. In: Molecular Immunology. 1982 ; Vol. 19, No. 9. pp. 1127-1138.
@article{bf79b5af48614e0fb15aea1585102c99,
title = "Characterization of antigens on human lymphocytes coded by messenger RNA translated in vitro",
abstract = "Translation and immunoprecipitation were used to identify messenger RNAs (mRNAs) coding for surface antigens expressed on human lymphoblastoid cells. The mRNAs were extracted from several human lymphoid cell lines as well as from fibroblastoid lines. These mRNAs were translated in vitro, and the translation products were reacted with xenoantisera raised against the antigens on human lymphoid cells. Products immunoprecipitated by these antisera were analysed by electrophoresis and fluorography. Four antisera immunoprecipitated a polypeptide with a mol. wt (MW) of approximately 32,000 (p32) from translations programmed with mRNA extracted from all the cell lines. Two antisera immunoprecipitated, in addition to p32. another polypeptide with a MW of approximately 25,000 (p25) only from translations programmed with RNA from lymphoid cell lines. p25 mRNA in the different lymphoid cell lines fell into three basic abundance classes as determined by in vitro translation and immunoprecipitation. Cells from two Burkitt's lymphomas (Raji and Daudi) did not express detectable p25 mRNA. Two T-lymphoblastoid lines (Molt-4 and 1301) contained five- to 10-fold less p25 mRNA than the B-lymphoid cell lines (Victor, RPMI-8866, RPMI-6410. RPM1-8226 and RPMI-1788). Both p32 and p25 were expressed on the cell surface inasmuch as lymphoblastoid cells adsorbed antibodies to both polypeptides. Human fibroblast, Raji or Daudi cells adsorbed anti-p32 antibodies from the antiserum but not anti-p25. Quantitative absorptions of the antiserum with T- or B-lymphoblastoid cells was used to determine the relative amounts of p32 and p25 expressed on the cell surface. B-Lymphoblastoid cells were found to express two- to five-fold more p25 on the cell surface than T-lymphoblastoid cells. p25 does not represent an immunoglobulin light-chain precursor inasmuch as a 1000-fold excess of unlabeled human Ig did not compete with p25 translated in vitro for binding by its respective antibody.",
author = "Robert Allen and Soldano Ferrone and Hoch, {James A.}",
year = "1982",
month = "1",
day = "1",
doi = "10.1016/0161-5890(82)90323-6",
language = "English",
volume = "19",
pages = "1127--1138",
journal = "Molecular Immunology",
issn = "0161-5890",
publisher = "Elsevier Ltd",
number = "9",

}

Characterization of antigens on human lymphocytes coded by messenger RNA translated in vitro. / Allen, Robert; Ferrone, Soldano; Hoch, James A.

In: Molecular Immunology, Vol. 19, No. 9, 01.01.1982, p. 1127-1138.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Characterization of antigens on human lymphocytes coded by messenger RNA translated in vitro

AU - Allen, Robert

AU - Ferrone, Soldano

AU - Hoch, James A.

PY - 1982/1/1

Y1 - 1982/1/1

N2 - Translation and immunoprecipitation were used to identify messenger RNAs (mRNAs) coding for surface antigens expressed on human lymphoblastoid cells. The mRNAs were extracted from several human lymphoid cell lines as well as from fibroblastoid lines. These mRNAs were translated in vitro, and the translation products were reacted with xenoantisera raised against the antigens on human lymphoid cells. Products immunoprecipitated by these antisera were analysed by electrophoresis and fluorography. Four antisera immunoprecipitated a polypeptide with a mol. wt (MW) of approximately 32,000 (p32) from translations programmed with mRNA extracted from all the cell lines. Two antisera immunoprecipitated, in addition to p32. another polypeptide with a MW of approximately 25,000 (p25) only from translations programmed with RNA from lymphoid cell lines. p25 mRNA in the different lymphoid cell lines fell into three basic abundance classes as determined by in vitro translation and immunoprecipitation. Cells from two Burkitt's lymphomas (Raji and Daudi) did not express detectable p25 mRNA. Two T-lymphoblastoid lines (Molt-4 and 1301) contained five- to 10-fold less p25 mRNA than the B-lymphoid cell lines (Victor, RPMI-8866, RPMI-6410. RPM1-8226 and RPMI-1788). Both p32 and p25 were expressed on the cell surface inasmuch as lymphoblastoid cells adsorbed antibodies to both polypeptides. Human fibroblast, Raji or Daudi cells adsorbed anti-p32 antibodies from the antiserum but not anti-p25. Quantitative absorptions of the antiserum with T- or B-lymphoblastoid cells was used to determine the relative amounts of p32 and p25 expressed on the cell surface. B-Lymphoblastoid cells were found to express two- to five-fold more p25 on the cell surface than T-lymphoblastoid cells. p25 does not represent an immunoglobulin light-chain precursor inasmuch as a 1000-fold excess of unlabeled human Ig did not compete with p25 translated in vitro for binding by its respective antibody.

AB - Translation and immunoprecipitation were used to identify messenger RNAs (mRNAs) coding for surface antigens expressed on human lymphoblastoid cells. The mRNAs were extracted from several human lymphoid cell lines as well as from fibroblastoid lines. These mRNAs were translated in vitro, and the translation products were reacted with xenoantisera raised against the antigens on human lymphoid cells. Products immunoprecipitated by these antisera were analysed by electrophoresis and fluorography. Four antisera immunoprecipitated a polypeptide with a mol. wt (MW) of approximately 32,000 (p32) from translations programmed with mRNA extracted from all the cell lines. Two antisera immunoprecipitated, in addition to p32. another polypeptide with a MW of approximately 25,000 (p25) only from translations programmed with RNA from lymphoid cell lines. p25 mRNA in the different lymphoid cell lines fell into three basic abundance classes as determined by in vitro translation and immunoprecipitation. Cells from two Burkitt's lymphomas (Raji and Daudi) did not express detectable p25 mRNA. Two T-lymphoblastoid lines (Molt-4 and 1301) contained five- to 10-fold less p25 mRNA than the B-lymphoid cell lines (Victor, RPMI-8866, RPMI-6410. RPM1-8226 and RPMI-1788). Both p32 and p25 were expressed on the cell surface inasmuch as lymphoblastoid cells adsorbed antibodies to both polypeptides. Human fibroblast, Raji or Daudi cells adsorbed anti-p32 antibodies from the antiserum but not anti-p25. Quantitative absorptions of the antiserum with T- or B-lymphoblastoid cells was used to determine the relative amounts of p32 and p25 expressed on the cell surface. B-Lymphoblastoid cells were found to express two- to five-fold more p25 on the cell surface than T-lymphoblastoid cells. p25 does not represent an immunoglobulin light-chain precursor inasmuch as a 1000-fold excess of unlabeled human Ig did not compete with p25 translated in vitro for binding by its respective antibody.

UR - http://www.scopus.com/inward/record.url?scp=0019963104&partnerID=8YFLogxK

U2 - 10.1016/0161-5890(82)90323-6

DO - 10.1016/0161-5890(82)90323-6

M3 - Article

C2 - 7144756

AN - SCOPUS:0019963104

VL - 19

SP - 1127

EP - 1138

JO - Molecular Immunology

JF - Molecular Immunology

SN - 0161-5890

IS - 9

ER -