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Re‐targeting of human lymphocytes expressing the T‐cell receptor gamma/delta to ovarian carcinoma cells by the use of bispecific monoclonal antibodies
Author(s) -
Ferrini Silvano,
Prigione Ignazia,
Mammouti Serafina,
Colnaghi Maria Ines,
Ménard Sylvie,
Moretta Alessandro,
Moretta Lorenzo
Publication year - 1989
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910440210
Subject(s) - monoclonal antibody , microbiology and biotechnology , t cell receptor , biology , t cell , ovarian carcinoma , antibody , polyclonal antibodies , cytolysis , gamma delta t cell , cancer research , immunology , ovarian cancer , cytotoxic t cell , immune system , cancer , biochemistry , in vitro , genetics
Abstract TcR gamma/delta + lymphocytes represent a small subset homogeneously composed of cytolytic T cells displaying unique motility and homing properties. Since the lytic machinery of these cells can be efficiently triggered by monoclonal antibodies (MAbs) directed to the TcR gamma/delta, such MAbs were used for the construction of bispecific MAbs in conjunction with an MAb specific for ovarian carcinoma cells. Hybrid hybridomas were obtained by fusing the Mov19 MAb (IgG 2a )‐producing hybridoma with either GI (IgG 2a ) or A13 (IgG 1 ) hybridomas, secreting MAbs specific for 2 peripheral blood subsets of TcR gamma/delta + lymphocytes. Hybrid hybridomas producing bispecific MAbs were screened according to their ability to induce ovarian carcinoma (IGROVI) target cell lysis by GI + or A13 + T cell clones, respectively. The Gl‐derived GM33.9 bispecific MAb induced selective lysis of Mov19 + ovarian carcinoma target cells only by GI + clones, whereas the A13‐derived AM 18.4 MAb was effective only in combination with A13 + clones. Neither the anti‐TcR gamma/ delta nor the Mov19 parental MAbs (used alone or in combination) induced target‐cell lysis. The hybrid nature (IgG 1 / lgG 2a ) of the AM 18.4 bispecific MAb was indicated by 2‐color immunofluorescence experiments. Thus, both ovarian carcinoma and A13 + effector cells were double stained by AM18.4 bispecific MAb followed by PE‐conjugated anti‐IgG 1 and FITC‐conjugated anti‐IgG 2a second reagents. Polyclonal TcR gamma/delta + cells were obtained by direct stimulation of peripheral blood mononuclear cells with Sepharose bead‐conjugated anti‐TcR gamma/delta MAbs and IL‐2. The lines so obtained contained more than 90% of TcR gamma/delta + cells after 4 weeks of culture, with an increase in TcR gamma/ delta + cell numbers ranging from 200 to 1,000‐fold. These TcR gamma/delta + cell lines efficiently lysed ovarian carcinoma target cells in the presence of bispecific MAb and may therefore represent a suitable source of effector cells for induction of ovarian carcinoma cell lysis.