Antigen-specific human T-cell hybridomas with helper activity.

DeFreitas, Elaine; Vella, Stefano; Linnenbach, Alban J.; Zmijewski, Chester M.; Koprowski, Hilary; Croce, Carlo M.

doi:10.1073/pnas.79.21.6646

Cited by 17 publications

(5 citation statements)

References 29 publications

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“…Because of the instability at the heterokaryon level immediately after the fusion, T-T cell hybrids producing single lymphokines or exhibiting single immunological functions can be generated (4,(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23). The major problem in developing T-T cell hybrids is the selection of the hybrids from the parent line.…”

Section: Resultsmentioning

confidence: 99%

A New Method for the Development of Human T-T Cell Hybrids Without the Use of Hat Medium

Platsoucas

Calvelli²,

Kunicka³

1987

Hybridoma

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We report here a new method for the development of human T-T cell hybrids by fusing mitogen- or alloantigen-stimulated T cells with non-mutagenized cells from human lymphoblastoid T cell lines. This method is based on a new selection procedure where the hybrids are separated from the parent T cell line on the basis of their ability to form colonies in soft agar. In contrast, cells from lymphoblastoid T cell lines Molt-4 and Jurkat do not form colonies in agar. Hybridoma colonies are retrieved from the agar plates, expanded in culture, screened by HLA-typing and appropriate functional tests and recloned several times by limiting dilution. HAT medium, which contains thymidine that appears to be toxic to the hybrids, is not used in our selection procedure. Using this method, we developed human T-T cell hybridomas (as determined by HLA-typing) producing B-cell growth factor (BCGF) either constitutively or after induction with Concanavalin A (Con A). Certain other T-T cell hybrids produced suppressor factor, significantly inhibiting proliferative responses of human peripheral blood mononuclear leukocytes to PHA, Con A and allogeneic cells in mixed lymphocyte culture.

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Section: Resultsmentioning

confidence: 99%

A New Method for the Development of Human T-T Cell Hybrids Without the Use of Hat Medium

Platsoucas

Calvelli²,

Kunicka³

1987

Hybridoma

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“…PBMCs were obtained from patients 1-5 before treatment and 1 month (patients 1 and 4), 5 months (patients 2 and 4), and 4 months (patient 5) after treatment, respectively. PBMCs were also obtained from two untreated patients (7 and 8) PBMCs from patients and healthy donors were stimulated in vitro in a modified Mishell-Dutton culture system as described in detail (12). Briefly, 6 x 105…”

Section: Methodsmentioning

confidence: 99%

“…PBMC Stimulation. PBMCs from patients and healthy donors were stimulated in vitro in a modified Mishell-Dutton culture system as described in detail (12). Briefly, 6 x 105…”

mentioning

confidence: 99%

Idiotypic cascades in cancer patients treated with monoclonal antibody CO17-1A.

Wettendorff

Iliopoulos

Tempero

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

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We have previously shown that gastrointestinal cancer patients treated with monoclonal antibody C017-1A (Abl) developed anti-idiotypic antibodies (Ab2) to the Abl. We now demonstrate that patients produce anti-anti-idiotypic antibodies (Ab3) to their autologous Ab2. Ab3 were demonstrated in culture supernatants of peripheral blood mononuclear cells from five Abl-treated patients after stimulation of the cells with heterologous Ab2 that functionally mimicked the tumor antigen (Ag) defined by Abl and immunologically cross reacted with the patients' Ab2. Ab3 shared idiotopes with AbM and were Abl-like in their binding specificities to tumor cells, Ag, and Ab2. Such antibodies were also elicited by stimulating cells with Ag. However, they were not produced by stimulating posttreatment mononuclear cells with control proteins or by stimulating pretreatment cells with either Ag or Ab2. Our results demonstrate idiotypic cascades in cancer patients treated with monoclonal antibody. Ag-specific Ab3 responses may underlie delayed clinical responses often observed in cancer patients treated with monoclonal antibodies of various specificities.We have previously demonstrated that gastrointestinal cancer patients treated with anti-colorectal carcinoma (CRC) monoclonal antibody (mAb) C017-1A, referred to hereafter as Abl, raised anti-idiotypic antibodies (Ab2) to the administered Abl. The Ab2 functionally mimicked in vitro the tumor antigen (Ag) defined by AbM (1-3). We had originally postulated that Ab2 induced in the patients anti-anti-idiotypic antibodies (Ab3) with Abl-like binding specificities for tumor cells (1). We had further postulated that Ab3 responses may underlie delayed clinical responses observed in some of the Abl-treated patients (1, 4). In the initial studies, Ab3 could not be demonstrated in patients' sera, presumably because they were bound to circulating Ab2 or to the patients' tumor cells. Therefore, in the present study, peripheral blood mononuclear cells (PBMCs) of Abl-treated patients were stimulated in culture with the Ag defined by AbM or with heterologous Ab2 that functionally mimics the Ag (5) and immunologically cross reacts with the patients' Ab2. The secreted human antibodies were characterized. Our results demonstrate idiotypic cascades in cancer patients treated with Abl. MATERIALS AND METHODSPatients. The clinical history and treatment plan of the five patients included in the present study have been described in detail (6). Briefly, five patients with advanced adenocarcinoma of the pancreas were treated with a single infusion of 400 mg of AbM mixed with autologous leukocytes. Two untreated patients (patients 7 and 8) with metastatic CRC also were included.Tumor Cells. Human CRC cells SW 1116 (7) and melanoma cells were used as target cells in mixed hemadsorption assay.Antibodies. Anti-CRC mAb C017-1A (IgG2a), referred to as Abl, has been described (7). As controls, anti-CRC mAb C029 (IgG1), anti-breast carcinoma mAb BR-140 (IgG2a), and anti-lung carcinoma mAb LU-16B13 (IgG2a) were u...

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“…Both populations were stained with F(ab')2 fragments of mAb 17-1A or anti-influenza mAb and goat anti-mouse fluorescein-conjugated Ig and subsequently analyzed in a cytofluorograph. In addition, peripheral blood mononuclear cells from the same patient were stimulated with F(ab')2 fragments of mAb 17-1A or anti-influenza mAb for 9 days in a modified MishellDutton culture for specific human Ig production (15). Supernatants from these cultures were assayed in a solid-phase enzyme-linked immunoabsorbent assay for specific human IgG (KPL Laboratories, Gaithersburg, MD).…”

mentioning

confidence: 99%

“…Peripheral blood mononuclear cells were stimulated with F(ab')2 fragments at 10 ng/ml of mAb 17-1A in vitro as described (15) to activate antigen-specific T cells. During the following 7 days, aliquots of cells were separated into T-and B-cell populations by rosetting with sheep erythrocytes treated with 2-aminoethylisouronium bromide (16).…”

mentioning

confidence: 99%

Human anti-idiotype antibodies in cancer patients: Is the modulation of the immune response beneficial for the patient?

Koprowski¹,

Herlyn²,

Lubeck³

et al. 1984

Proc. Natl. Acad. Sci. U.S.A.

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211

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Inoculation of human subjects with mouse monoclonal antibody results in the production of anti-idiotype antibody that reacts with the binding site of the monoclonal antibody. This reaction is hapten-inhibited, suggesting that an internal image of the antigen is produced by the anti-idiotype response. The anti-idiotype antibody isolated from sera of three patients showed significant crossreactivity. Patients who developed the anti-idiotype antibody improved clinically and had long remission from their disease. The possible presence of the internal image of cancer antigen on the human immunoglobulin molecule may change the conditions under which the immune system reacts to the tumor antigen and may open new approaches to the control of tumor growth.

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Antigen-specific human T-cell hybridomas with helper activity.

Cited by 17 publications

References 29 publications

A New Method for the Development of Human T-T Cell Hybrids Without the Use of Hat Medium

A New Method for the Development of Human T-T Cell Hybrids Without the Use of Hat Medium

Idiotypic cascades in cancer patients treated with monoclonal antibody CO17-1A.

Human anti-idiotype antibodies in cancer patients: Is the modulation of the immune response beneficial for the patient?

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