The Journal of Immunology, Vol 131, Issue 1 334-340, Copyright © 1983 by American Association of Immunologists

ARTICLES

Characterization of a novel differentiation antigen complex recognize by a monoclonal antibody (A-1A5): unique activation-specific molecular forms on stimulated T cells

ME Hemler, CF Ware and JL Strominger

By using a single monoclonal antibody, a novel glycoprotein complex composed of at least three distinct bands was defined on the surface of mitogen- or alloantigen-stimulated T cells. These bands (210,000, 165,000, and 130,000 Mr) were not disulfide linked and could be radio- labeled with 125I or 35S-methionine and readily detected by immunoprecipitation with the monoclonal antibody A-1A5. Only approximately 20% of normal T cells (E rosette positive) and approximately 47% non-T cells (E rosette negative) were reactive with A- 1A5. However, upon activation of T cells, the amount of A-1A5 binding per cell and the percentage of positive cells significantly increased. This increase was most pronounced in the activated cell subpopulations currently undergoing cell division (S, G2, and M phases), which became 79% A-1A5 positive (after PHA stimulation) and 99% A-1A5 positive (in long-term culture with alloantigen and IL2). Resting lymphocytes contained only the 130,000 Mr band reactive with A-1A5. The two other bands (210,000 and 165,000 Mr) were markers for T cell activation and only appeared several days after T cell stimulation and became especially prominent after the addition of exogenous IL 2. All T lymphoblastoid cell lines tested expressed at least the lower bands (130,000 Mr), and the T cell line HSB also expressed one of the activation-related larger proteins (165,000 Mr). B lymphoblastoid cell lines expressed only a very weak lower band (130,000 Mr), and the cell line U-937 (in the monocyte-macrophage lineage) expressed only a single band (145,000 Mr) not aligned with any of the bands found on lymphoid cells. The estimated number of A-1A5 binding sites per cell was much higher on U-937 (11 X 10(5)), and generally higher on other cell lines of myeloid lineage (1 to 4 X 10(5)) than on lymphoid cell lines (0.2 to 1.3 X 10(5)).

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