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  • 1
    ISSN: 1432-0851
    Keywords: Key words Apoptosis ; Antibody-dependent cell-mediated cytotoxicity ; Monocyte ; 17-1A ; Interferon γ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  Antibody-dependent cell-mediated cytotoxicity (ADCC) has been considered to be one of the main effector mechanisms by which unconjugated monoclonal antibody (mAb) 17-1A can exert an antitumor effect in vivo. Since the apoptotic pathway as well as the necrotic pathway have been shown to be utilized in various cytotoxic effector mechanisms, we investigated the role of apoptosis in ADCC mediated by monocytes (ADMC) using mAb 17-1A as an antibody and the human colorectal carcinoma cell line, COLO205, as target cells in vitro. The implications of the apoptosis during ADMC was demonstrated by means of both a DNA fragmentation assay and a TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay. Furthermore, interferon γ (IFNγ) was also found to enhance the induction of apoptosis significantly. The addition of superoxide dismutase did not reduce the level of the apoptosis, although superoxide anion (O2 –) was observed to be produced. However, the release of tumor necrosis factor α (TNFα) was significantly enhanced during ADMC, while, in addition, apoptosis was significantly inhibited by the addition of anti-TNFα antibody. These findings indicated that apoptosis might be implicated in ADMC with mAb 17-1A, which was augmented by IFNγ, while, in addition, TNFα may also be one of the major mediators of apoptosis.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0851
    Keywords: Key words Cytokine ; Quantitative RT-PCR ; Interleukin-2 ; Interleukin-1 ; Tumor necrosis factor α
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  After activation with interleukin-2 (IL-2), peripheral blood mononuclear cells (PBMC) have been reported to induce the expression of mRNA coding various cytokines, including interleukin(IL)-1α, -1β and tumor necrosis factor α (TNFα). We examined the cytokine mRNA expression of PBMC following treatment with IL-2 in vitro and in vivo by a quantitative method using the reverse transcription/polymerase chain reaction (RT-PCR). After stimulating PBMC with IL-2 in vitro, peak levels of IL-1α mRNA were reached between 3 h and 12 h, and thereafter declined. The IL-1β expression increased, with levels peaking at 1–6 h and, had decreased by 96 h. The expression of TNFα was elevated both 1–3 h and 24–48 h after stimulation. The peak levels of IL-1α and -1β mRNA and the early elevation of TNFα mRNA mainly accounted for the cytokine mRNA expression in adherent cells; however, the late induction of TNFα mRNA was observed in nonadherent cells. In patients with advanced carcinoma, the IL-1α and -1β mRNA expression were elevated after IL-2 treatment for 5 consecutive days, while the expression of TNFα mRNA also increased. These results indicate that the quantitative RT-PCR method appears to be useful for analyzing the cytokine mRNA expression of PBMC after treatment with IL-2.
    Type of Medium: Electronic Resource
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