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Transformation-associated 86 kDa natural killer target molecule expressed on the mouse, rat and human cell surface
1Department of Pathology, Sapporo Medical University School of Medicine.
Abstract:
We previously reported on the 86 kDa natural killer target molecule associated with transformation of the oncogene-transfected rat fibroblasts. This molecule may participate in the lethal hit phase of cytotoxicity by natural killer (NK) cells. Originally, this molecule was defined by mAb109, but mAb109 could react only with rat tumor lines. In this report, to determine whether the 86 kDa molecule could be utilized as a natural killer target molecule in mammalian cells, we developed a polyclonal anti-86 kDa antibody (pAb109). Our data indicated that pAb109 preferentially reacted with NK-susceptible lines such as mouse YAC-1, rat W31 and human K562 cells, but reacted only weakly with NK-resistant mouse EL-4, rat fetal fibroblast WFB and human fetal fibroblast HEPM. In a cytotoxicity experiment, pAb109 F(ab')2 fragments could inhibit the cytolysis by NK cells of W31 and K562 cells. However, these fragments did not inhibit the cytotoxicity of non-NK cells such as CD3+, CD4-, CD8- T cell receptor alpha beta- T cells (presumably gamma delta T cells) to W31 cells. Taken together, these data suggest that the cell transformation-associated 86 kDa molecule may be critical in NK cytotoxicity, and a candidate for the NK target structure in mammalian tumor cells.
Insights
A newly developed antibody confirms the 86 kDa molecule is a key target for natural killer (NK) cells in mammalian tumors. This finding advances understanding of tumor cell killing by NK cells.
Area of Science:
- Immunology
- Cell Biology
- Oncology
Background:
- A previously identified 86 kDa molecule is associated with oncogene-induced transformation in rat fibroblasts.
- This molecule may play a role in the lethal hit phase of natural killer (NK) cell-mediated cytotoxicity.
- Initial characterization using mAb109 was limited to rat tumor lines.
Purpose of the Study:
- To determine if the 86 kDa molecule functions as a natural killer (NK) target molecule in various mammalian cells.
- To develop a more broadly reactive antibody against the 86 kDa molecule.
- To investigate the role of the 86 kDa molecule in NK cell cytotoxicity.
Main Methods:
- Development of a polyclonal anti-86 kDa antibody (pAb109).
- Testing antibody reactivity against NK-susceptible and NK-resistant cell lines (mouse, rat, human).
- Cytotoxicity assays using pAb109 F(ab')2 fragments to inhibit NK cell-mediated lysis.
Main Results:
- pAb109 demonstrated preferential reactivity with NK-susceptible cell lines (YAC-1, W31, K562) compared to NK-resistant lines (EL-4, WFB, HEPM).
- pAb109 F(ab')2 fragments inhibited NK cell-mediated cytotoxicity against W31 and K562 cells.
- These fragments did not inhibit cytotoxicity mediated by non-NK T cells (gamma delta T cells) against W31 cells.
Conclusions:
- The 86 kDa molecule, associated with cell transformation, is a critical component in NK cell-mediated cytotoxicity.
- This molecule is a potential target structure on mammalian tumor cells for NK cell recognition and killing.
- The findings support the role of the 86 kDa molecule in NK cell-mediated tumor cell elimination.
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