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  • Title: Embryonic stem cells and embryoid bodies express lymphocyte costimulatory molecules.
    Author: Ling V, Munroe RC, Murphy EA, Gray GS.
    Journal: Exp Cell Res; 1998 May 25; 241(1):55-65. PubMed ID: 9633513.
    Abstract:
    Despite the importance of the costimulatory proteins B7-1 (CD80), B7-2 (CD86), and their counterreceptors CD28 and CTLA-4 (CD154) in the regulation of T cell proliferation in the adult immunological system, the initial appearance of these proteins during embryonic development has not been investigated. Using in vitro cultures of undifferentiated mouse embryonic stem (ES) cells and differentiating embryoid bodies as a model of very early embryonic development, we examined these cells for the presence of mRNA and protein corresponding to the B7 and CD28 families of costimulatory molecules. By flow cytometry, a stochastically regulated subpopulation of B7-1+ cells comprising 33% of total cells was detected in ES cell cultures, while negligible staining was found for B7-2, CTLA-4, and CD28. When ES cells were differentiated into embryoid bodies for 12 days, a CD45+ subpopulation of embryoid body cells were found to stain positively for B7-1, B7-2, and CD28. RT-PCR confirmed cell staining data by revealing amplification products corresponding to B7-1, B7-2, and CD28 in corresponding samples. Very low levels of CTLA-4 amplification products were found in all samples; however, surface staining of CTLA-4 was never detected. The functional capacity of ES cell B7-1 to bind its ligand was verified by the ability of the soluble fusion protein CTLA-4-Ig to bind ES cells and the ability of this reagent to block anti-B7-1 antibody binding in cell based competition assays. These results demonstrate that expression of costimulatory molecules arises very early during in vitro development and suggests that the early embryonic environment may utilize cellular signaling systems analogous to those seen in the immune system.
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