Biochemical characterization of Ia alloantigens in guinea pigs. II. Comparative peptide mapping of Ia antigens from B cells, T cells, and Mφ

C. R. Lyons, R. G. Cook, T. F. Tucker, J. W. Uhr

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Abstract

Radioactive Ia.4 molecules were prepared from 3H- or 14C-labeled splenocytes, selected PEL, or bronchoalveolar macrophages (Mφ). Studies in the accompanying paper indicated that incorporation into Ia.4 in these 3 populations is due to B cells, T cells, and Mφ, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to isolate the 58,000 m.w. form of Ia.4. The eluted molecule was then reduced, and the resultant α and β subunits were isolated by a second separation on SDS-PAGE. α (or β) chains frowm 1 population labeled with 3H-amino acids was mixed with α (or β) chains obtained from a population representing a different cell type that was labled with 14C-amino acids and the mixture was digested with trypsin. Double-label (3H/14C) comparative peptide mapping was performed using high-pressure liquid chromatography to separate the peptides. Eighteen to 20 peaks of radioactivity were resolved from α chains, and 12 to 15 from β chains. No reproducible differences were observed when comparing α or β chains of T cells and Mφ, those of B cells and Mφ. These results indicate that the primary structure of Ia.4 molecules is identical on the 3 cell types in question. The implications of having a T cell bearing the same Ia that it recognizes on a Mφ in conjunction with antigen is discussed.

Original languageEnglish (US)
Pages (from-to)1885-1888
Number of pages4
JournalJournal of Immunology
Volume127
Issue number5
StatePublished - 1981

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Peptide Mapping
Isoantigens
Histocompatibility Antigens Class II
Guinea Pigs
B-Lymphocytes
T-Lymphocytes
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Population
Amino Acids
Trypsin
Radioactivity
Macrophages
High Pressure Liquid Chromatography
Antigens
Peptides

ASJC Scopus subject areas

  • Immunology

Cite this

Biochemical characterization of Ia alloantigens in guinea pigs. II. Comparative peptide mapping of Ia antigens from B cells, T cells, and Mφ. / Lyons, C. R.; Cook, R. G.; Tucker, T. F.; Uhr, J. W.

In: Journal of Immunology, Vol. 127, No. 5, 1981, p. 1885-1888.

Research output: Contribution to journalArticle

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