Limits...
Redirecting mouse T hybridoma against human breast and ovarian carcinomas: in vivo activity against HER-2/neu expressing cancer cells.

Gritzapis AD, Mamalaki A, Kretsovali A, Papamatheakis J, Belimezi M, Perez SA, Baxevanis CN, Papamichail M - Br. J. Cancer (2003)

Bottom Line: The scFv(anti-HER-2/neu)/zeta chimeric gene was successfully expressed as a functional surface receptor in the MD.45 CTL hybridoma (MD.45-HER/zeta).The MD.45-HER/zeta-transduced cells also lysed HER-2/neu(+) target cells in vitro with high specificity.The adoptively transferred MD.45-HER/zeta cells both slowed significantly the growth of human FM3 melanoma or murine ALC leukaemic cells both transfected to express HER-2/neu.

View Article: PubMed Central - PubMed

Affiliation: Saint Savas Cancer Hospital, Cancer Immunology and Immunotherapy Center, Athens, Greece.

ABSTRACT
Chimeric receptors comprising of the T-cell receptor-zeta cytoplasmic signalling chain fused to an extracellular ligand-binding domain of a single-chain antibody (scFv) have served as effective tools for redirecting cytotoxic T lymphocytes (CTL) against tumour cells. In this report, we constructed a chimeric scFv/zeta gene composed of the variable regions of an HER-2/neu-specific monoclonal antibody (MAb) joined to the TCR-zeta chain. The scFv(anti-HER-2/neu)/zeta chimeric gene was successfully expressed as a functional surface receptor in the MD.45 CTL hybridoma (MD.45-HER/zeta). More importantly, the scFv(anti-HER-2/neu)/zeta receptor was functionally active, since it triggered cytokine secretion by the MD.45-HER/zeta cells upon recognition of HER-2/neu-positive (+) tumour cell lines, or primary tumour cells from patients with HER-2/neu(+) cancers. The MD.45-HER/zeta-transduced cells also lysed HER-2/neu(+) target cells in vitro with high specificity. We tested the antitumour efficacy of scFv(anti-HER-2/neu)/zeta expressing MD.45 cells in severe combined immunodeficiency disease mice/human and murine tumour models. The adoptively transferred MD.45-HER/zeta cells both slowed significantly the growth of human FM3 melanoma or murine ALC leukaemic cells both transfected to express HER-2/neu. Our data demonstrate the feasibility of redirecting MD.45 CTL with the scFv(anti-HER-2/neu)/zeta chimeric receptor to respond specifically against HER-2/neu expressing tumour cells in vitro and in vivo. Moreover, they make it likely that T cells transduced with the same chimeric gene might be utilised in the treatment of patients with HER-2/neu(+) tumours.

Show MeSH

Related in: MedlinePlus

Survival of SCID mice after subcutaneous injection (day 1) with human melanoma FM3 (FM3-HER, FM3-mock) or murine leukaemic ALC (ALC-HER, ALC-mock) cell lines. On the same day (day 1) and for the following 2 days (days 2 and 3) mice were treated with PBS or MD45 cells transduced either with scFv(anti-HER-2/neu)/ζ (MD.45-HER/ζ) or with their mock tranfectants (MD.45-mock). In all cases, mice treated with MD.45-HER/ζ demonstrated a significant increase in survival compared to the other groups (P<0.01).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2747561&req=5

fig5: Survival of SCID mice after subcutaneous injection (day 1) with human melanoma FM3 (FM3-HER, FM3-mock) or murine leukaemic ALC (ALC-HER, ALC-mock) cell lines. On the same day (day 1) and for the following 2 days (days 2 and 3) mice were treated with PBS or MD45 cells transduced either with scFv(anti-HER-2/neu)/ζ (MD.45-HER/ζ) or with their mock tranfectants (MD.45-mock). In all cases, mice treated with MD.45-HER/ζ demonstrated a significant increase in survival compared to the other groups (P<0.01).

Mentions: To assess whether MD.45-HER/ζ cells had significant in vivo activity against HER-2/neu+ tumour cells, 1 × 106 murine ALC-HER lymphoma cells or an equal number of human FM3-HER-melanoma cells were inoculated s.c. in mice that had been irradiated (200 rad) 1 day before to suppress endogenous NK activity (Dorshkind et al, 1985; Lu et al, 1994). On the same day with tumour cell inoculation and for the following 2 days, mice were treated with i.p. injections (one injection per day) of MD.45-HER/ζ or MD.45-mock-transduced cells. Mice treated with MD.45-HER/ζ cells experienced a significantly prolonged survival as compared to those treated with MD.45-mock cells (for both models >100 days; P<0.001) (Figure 5Figure 5


Redirecting mouse T hybridoma against human breast and ovarian carcinomas: in vivo activity against HER-2/neu expressing cancer cells.

Gritzapis AD, Mamalaki A, Kretsovali A, Papamatheakis J, Belimezi M, Perez SA, Baxevanis CN, Papamichail M - Br. J. Cancer (2003)

Survival of SCID mice after subcutaneous injection (day 1) with human melanoma FM3 (FM3-HER, FM3-mock) or murine leukaemic ALC (ALC-HER, ALC-mock) cell lines. On the same day (day 1) and for the following 2 days (days 2 and 3) mice were treated with PBS or MD45 cells transduced either with scFv(anti-HER-2/neu)/ζ (MD.45-HER/ζ) or with their mock tranfectants (MD.45-mock). In all cases, mice treated with MD.45-HER/ζ demonstrated a significant increase in survival compared to the other groups (P<0.01).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2747561&req=5

fig5: Survival of SCID mice after subcutaneous injection (day 1) with human melanoma FM3 (FM3-HER, FM3-mock) or murine leukaemic ALC (ALC-HER, ALC-mock) cell lines. On the same day (day 1) and for the following 2 days (days 2 and 3) mice were treated with PBS or MD45 cells transduced either with scFv(anti-HER-2/neu)/ζ (MD.45-HER/ζ) or with their mock tranfectants (MD.45-mock). In all cases, mice treated with MD.45-HER/ζ demonstrated a significant increase in survival compared to the other groups (P<0.01).
Mentions: To assess whether MD.45-HER/ζ cells had significant in vivo activity against HER-2/neu+ tumour cells, 1 × 106 murine ALC-HER lymphoma cells or an equal number of human FM3-HER-melanoma cells were inoculated s.c. in mice that had been irradiated (200 rad) 1 day before to suppress endogenous NK activity (Dorshkind et al, 1985; Lu et al, 1994). On the same day with tumour cell inoculation and for the following 2 days, mice were treated with i.p. injections (one injection per day) of MD.45-HER/ζ or MD.45-mock-transduced cells. Mice treated with MD.45-HER/ζ cells experienced a significantly prolonged survival as compared to those treated with MD.45-mock cells (for both models >100 days; P<0.001) (Figure 5Figure 5

Bottom Line: The scFv(anti-HER-2/neu)/zeta chimeric gene was successfully expressed as a functional surface receptor in the MD.45 CTL hybridoma (MD.45-HER/zeta).The MD.45-HER/zeta-transduced cells also lysed HER-2/neu(+) target cells in vitro with high specificity.The adoptively transferred MD.45-HER/zeta cells both slowed significantly the growth of human FM3 melanoma or murine ALC leukaemic cells both transfected to express HER-2/neu.

View Article: PubMed Central - PubMed

Affiliation: Saint Savas Cancer Hospital, Cancer Immunology and Immunotherapy Center, Athens, Greece.

ABSTRACT
Chimeric receptors comprising of the T-cell receptor-zeta cytoplasmic signalling chain fused to an extracellular ligand-binding domain of a single-chain antibody (scFv) have served as effective tools for redirecting cytotoxic T lymphocytes (CTL) against tumour cells. In this report, we constructed a chimeric scFv/zeta gene composed of the variable regions of an HER-2/neu-specific monoclonal antibody (MAb) joined to the TCR-zeta chain. The scFv(anti-HER-2/neu)/zeta chimeric gene was successfully expressed as a functional surface receptor in the MD.45 CTL hybridoma (MD.45-HER/zeta). More importantly, the scFv(anti-HER-2/neu)/zeta receptor was functionally active, since it triggered cytokine secretion by the MD.45-HER/zeta cells upon recognition of HER-2/neu-positive (+) tumour cell lines, or primary tumour cells from patients with HER-2/neu(+) cancers. The MD.45-HER/zeta-transduced cells also lysed HER-2/neu(+) target cells in vitro with high specificity. We tested the antitumour efficacy of scFv(anti-HER-2/neu)/zeta expressing MD.45 cells in severe combined immunodeficiency disease mice/human and murine tumour models. The adoptively transferred MD.45-HER/zeta cells both slowed significantly the growth of human FM3 melanoma or murine ALC leukaemic cells both transfected to express HER-2/neu. Our data demonstrate the feasibility of redirecting MD.45 CTL with the scFv(anti-HER-2/neu)/zeta chimeric receptor to respond specifically against HER-2/neu expressing tumour cells in vitro and in vivo. Moreover, they make it likely that T cells transduced with the same chimeric gene might be utilised in the treatment of patients with HER-2/neu(+) tumours.

Show MeSH
Related in: MedlinePlus