Locoregional immunotherapy of malignant effusion from colorectal cancer using the streptococcal preparation OK-432 plus Interleukin-2

Abstract

In total, 16 patients with cytologically proven malignant effusion from colorectal cancer were treated by locoregional administration of the streptococcal preparation OK-432 alone or OK-432 plus the T-cell growth factor interleukin (IL)-2, and the action mechanism of the treatment was studied. A positive clinical response, showing a cytologic disappearance of cancer cells and decrease of effusion, was observed in nine of 11 (82%) patients treated with OK-432 alone and in all five patients treated with OK-432 plus IL-2. Flow cytometric analysis revealed that OK-432 plus IL-2 locally induced acute inflammation-like responses, including serial cellular infiltrations of granulocyte migration within a matter of hours, and activation of macrophages and T lymphocyte involvement within the following days, and that a predominant expansion of CD3 þ CD4 þ lymphocytes (CD: cluster of differentiation) was induced by in vitro stimulation with IL-2 of locoregional cells after the OK-432 administration (OK/IL-2AK cells). The OK/IL-2AK cells produced tumour necrosis factor-a and interferon-g, but these cells did not produce IL-4 and IL-6. The OK/IL-2AK cells expressed potent killing activity against autologous tumour cells. This activity was abrogated by treatment of the lymphocytes with anti-CD3, -CD4, -TCRab antibody, and by the treatment of target cells with anti-human leukocyte antigen (HLA)-DR antibody. The OK/IL-2AK cells expressed Fas-L gene, and flow cytometric analysis demonstrated HLA-DR expression in approximately 75% of CEA þ or cytokeratin þ effusion cells. TCRVb gene analysis of the OK/IL-2AK cells showed an oligoclonal usage of TCRb20, which was also involved in the cytotoxic mechanism of the OK/IL-2AK cells. Single-strand conformational polymorphism analysis demonstrated the clonotypes for the TCRVb20 gene, and the CDR3s of the gene were sequenced. The clonotypic PCR using the TCRVb20-CDR3 sequences could detect the CDR3-identical TCRs in effusion lymphocytes from the other patients. Taken together, it is suggested that locoregional administration of OK-432 plus IL-2 is highly effective for the management of malignant effusion from colorectal cancer. OK-432 plus IL-2 induces autologous tumour-reactive CD4 þ Th1 killer lymphocytes, which recognise tumour antigen(s) presented with HLA class II molecules on effusion tumour cells by means of preferential usage of TCRVb20. The clonotypic PCR using the TCRVb20-CDR3 sequences may be informative for treating malignant effusion from colorectal cancer using OK-432 plus IL-2.

OK432-Activated Human Dendritic Cells Kill Tumor Cells via CD40/CD40 Ligand Interactions

“…Tumor cell killing by OK-DC may be particularly suitable for clinical application and development, as OK432 has been safely administered to cancer patients for many years (35,36).…”

OK-432 Suppresses Proliferation and Metastasis by Tumor Associated Macrophages in Bladder Cancer

“…OK-432, which is a penicillin-killed and lyophilized preparation of a low-virulence strain of Streptococcus pyogenes (group A), has been successfully used as an immunotherapeutic agent against many types of malignancies, including head and neck cancer, stomach cancer, lung cancer, pancreatic cancer and colorectal cancer (Yamaguchi et al, 2003;Yoo et al, 2010;Nakamoto et al, 2011;Tano et al, 2012;Tano et al, 2013;Wada et al, 2014). It has been reported that OK-432 elicits anti-tumor effects by stimulating immunocompetent cells such as macrophages, T cells, and natural killer (NK) cells, and by inducing helper T-cell 1(Th1) type cytokines, such as IFN-γ, TNF-α, IL-6, IL-8, IL-10, IL-12, and IL-18(Oshimi et al, 1980;Saito et al, 1982;Fujimoto et al, 1997), which can augment cytotoxic T lymphocytes to antitumor.…”

Novel Intrapleural Therapies for Malignant Diseases

“…These include inactivated bacterial superantigens from streptococcus (OK-432) or staphylococcus, which were studied in combination with other treatment modalities. Yamaguchi et al [34] combined OK-432 with IL-2 into the pleural space in 16 patients with documented MPE from colorectal cancer. There was a cytologic response and decrease in effusion volume in 9 of 11 (82%) patients treated with OK-432 alone and in all 5 patients treated with OK-432 plus IL-2.…”