TY - JOUR
T1 - Characterization of antigens on human lymphocytes coded by messenger RNA translated in vitro
AU - Allen, Robert W.
AU - Ferrone, Soldano
AU - Hoch, James A.
N1 - Funding Information:
Acicfloizird~cm~11-rTsh e authors express their thanks to Kenton Slaven for excellent technical assistance. R.W.A. is the recipient of a post-doctoral fellowship No. PF1486 from the American Cancer Society. This research was supported, in part. by grants GM 19516,G M 25891, AI 19189 and CA 28555 awarded by the National Institute of General Medical Sciences. PHSDHHS, the National Cancer Institute PHSIDHHS.
PY - 1982/9
Y1 - 1982/9
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
SN - 0161-5890
VL - 19
SP - 1127
EP - 1138
JO - Molecular Immunology
JF - Molecular Immunology
IS - 9
ER -