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We have analyzed the transmembrane signaling operating in human cytolytic lymphocytes lacking surface expression of the CD3/TCR complex. Peripheral blood lymphocytes were fractionated into CD3+ and CD3- on the FACS and cloned under limiting conditions in the presence of PHA and IL-2. Approximately 90% CD3+ and 10% CD3- cells underwent clonal expansion. Clones obtained from the CD3- fraction belonged to two main phenotypic groups: CD2+ CD7+ and CD2- CD7+. Several clones were expanded and analyzed for surface phenotype and function. All of the five clones selected for detailed analysis did not express CD4, CD8, and CD28 antigens and did not release IL-2, whereas they displayed cytolytic activity against NK-sensitive, NK-resistant, and fresh tumor target cells. After stimulation with anti-CD2 mAbs or PHA a rapid increase in [Ca2+]i was detected in CD3- CD2+ CD7+ clones. This increment was caused by the release of Ca2+ from intracellular stores and by the influx from the extracellular compartment. Signaling in response to PHA did not appear to be dependent upon surface expression of CD2 molecules since antibody-induced modulation of CD2 did not prevent PHA-induced signal transduction. Similarly, in CD3- CD2- CD7+ clones [Ca2+]i increments and inositol phosphate formation occurred after stimulation with PHA. These data indicate that the functional PHA-binding structures, expressed in both groups of CD3- clones, are distinct from CD3/TCR complex and CD2 molecules.

the journal of experimental medicine/the journal of experimental medicine

Human cytolytic cell clones lacking surface expression of T cell receptor alpha/beta or gamma/delta. Evidence that surface structures other than CD3 or CD2 molecules are required for signal transduction.