Interleukin 2 restores CD3-ζ chain expression but fails to generate tumour-specific lytic activity in tumour-infiltrating lymphocytes derived from human colorectal hepatic metastases

Abstract

Summary Metastatic colorectal cancer is usually progressive despite infiltration of the tumours by T lymphocytes, suggesting that these tumour-infiltrating lymphocytes (TILs) are functionally deficient. Recently, TILs from other tumours have been shown to express reduced levels of the T-cell receptor signal-transducing CD3-; chain. We were interested to determine whether a similar abnormality existed in TILs from human colorectal hepatic metastasis (CHM) and, if so, whether correcting the abnormality in vitro would restore anti-tumour activity and provide support for the development of immunotherapy for colorectal hepatic metastases. Twelve of 19 TILs from colorectal hepatic metastases were successfully expanded in vitro in high-dose recombinant interleukin 2 (rlL-2) and their specific anti-tumour cytolytic activity was determined. CD3-positive (CD3+) TILs were HLA-Ddbsh and CD69high, suggesting that they had been activated by exposure to antigen but expressed low levels of CD25, CD71 and the nuclear proliferation antigen Ki-67. Furthermore, they showed reduced expression of CD3-4 compared with autologous peripheral blood T cells (PBTs) and failed to proliferate in the absence of high-dose rlL-2. Expansion of TILs in rlL-2 resulted in restoration of CD3-t expression and the ability to lyse K562 and Daudi cells but not autologous tumour cells. The absence of autologous tumour-specific cytolytic T-cell (CTL) activity may be due to the poor immunogenicity of colorectal tumour cells, which we found expressed only low levels of MHC antigens and CD54 and failed to express MHC II antigens or the co-stimulatory molecules CD80, CD86 or CD106. The inability of rlL-2 to generate tumour-specific CTLs despite restoration of CD3-l expression and the presence of an intact lytic mechanism suggests that successful immunotherapy may require the development of strategies to increase the immunogenicity of this tumour.

T lymphocytes isolated from patients with advanced colorectal cancer are suitable for gene immunotherapy approaches

“…In the light of published work demonstrating the dysfunctional properties of T lymphocytes isolated from patients with a cancer load (Rosenberg et al, 1994;Bukowski et al, 1998;Yoong and Adams, 1998), the aims of this project were to assess whether the conditions used to activate and transduce T lymphocytes from normal donors were equally effective in T lymphocytes from patients with advanced colorectal cancer. In addition, it is important to evaluate whether CIR crosslinking in the presence of antigen would generate a response in patient cells similar to that of normal donors.…”

“…T lymphocytes isolated from patients with advanced melanoma (Saito et al, 2000), and head and neck cancer (Hoffman et al, 2002) have also been shown to be more susceptible to spontaneous apoptosis thereby potentially compromising the patient's own immune competence. Furthermore, tumour-infiltrating lymphocytes isolated from colorectal hepatic metastases have been shown to have reduced CD3z chain expression and, upon expansion, lack tumour-specific lytic activity (Yoong and Adams, 1998). In view of this work and to further determine the viability of our approach in a clinical setting, T lymphocytes were isolated from the peripheral blood of patients with advanced colorectal disease (hepatic colorectal metastases) in order to determine whether they could be efficiently gene modified to express a chimeric immune receptor.…”

Expression and function of CXC and CC chemokines in human malignant liver tumors: A role for human monokine induced by ?-interferon in lymphocyte recruitment to hepatocellular carcinoma

“…Thus, the ability of HCC to recruit circulating lymphocytes suggests that immunotherapy should have a higher chance of success in these tumors than in colorectal hepatic metastases, that fail to support lymphocyte recruitment. 41,64 …”

A Novel Thymoma-Associated Immunodeficiency with Increased Naive T Cells and Reduced CD247 Expression

“…Thus, the neoplastic thymic epithelium might disturb the T cell development causing both an increased output of gd T cells and reduced amounts of CD247 in the TCRs of maturing ab and gd T cells through excessive stimulation of developing T lymphocytes. This would explain the reversibility of the CD247 defect in our patient after prolonged T cell stimulation in vitro, as has already been observed in the case of tumor-infiltrating lymphocytes (39). Additionally, it could explain the unusual association of higher numbers of naive T cells in the periphery (4,52) with the development of thymoma-related autoimmune conditions such as myasthenia gravis (47) and giant cell myocarditis (1,2).…”

“…All TCR alterations in patient B were restored following prolonged T cell activation by PHA-P and IL-2 (Fig. 4A, 4B), which parallels the restoration of CD247 expression in tumor-infiltrating lymphocytes by IL-2 (39). Thus, these abnormalities were most likely acquired rather than genetically determined (40), as also indicated by the lack of mutations in the CD247 (CD3Z) gene of patient B.…”