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Jung, G.* ; Freimann, U. ; von Marschall, Z.* ; Reisfeld, R.A.* ; Wilmanns, W.

Target cell-induced T cell activation with bi- and trispecific antibody fragments.

Eur. J. Immunol. 21, 2431-2435 (1991)

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Previously we proposed a concept for tumor immunotherapy in which two different bispecific antibody conjugates, an anti-target x anti-CD3 and an anti-target x anti-CD28 conjugate, induce the activation of resting humanTcells upon binding to the respective tumor target cells. After in vivo application of these reagents, this model of a "target cell-induced T cell activation" envisages the destruction of target cells by in situ activated T cells. Obviously however, for in vivo application, the use of Fc-free antibody fragments is mandatory to prevent binding of the conjugates to Fc receptor-positive cells which would lead to Fc-mediated T cell activation. Here we report a simplification of published procedures for the generation of bispecific Fab-hybrid fragments, univalent for each antigen. We demonstrate that an anti-target x anti-CDS/anti-target X antiCD28 combination of such hybrids, as well as an identical combination of covalently coupled F(ab′)2 fragments, mediate "target cell-induced Tcell activation" in an in vitro test system.Thus, these reagents may be capable of inducing an in situ activation of human T cells upon systemic in vivo application according to the concept outlined above. A trispecific conjugate with anti-target, anti-CD3-and anti-CD28 specificity appears to be unsuitable for this purpose because it activates resting T cells in soluble form without requiring immobilization through binding via its anti-target portion.

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