Glycoproteic nature of surface molecules of effector cells with lymphokine-activated killer (LAK) activity. Evidence that T11, T8 or T3 molecules are not involved in tumor-cell lysis by LAK effector T cells
✍ Scribed by Marcello Bagnasco; Maria Cristina Mingari; Daniela Pende; Daniela Venuti; Giorgio Walter Canonica; Lorenzo Moretta
- Publisher
- John Wiley and Sons
- Year
- 1987
- Tongue
- French
- Weight
- 506 KB
- Volume
- 39
- Category
- Article
- ISSN
- 0020-7136
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✦ Synopsis
Human peripheral blood mononuclear cells cultured in the presence of interleukin-2 (IL-2) acquire the capability of lysing NK-resistant fresh tumor target cells. in an attempt to (leiin-Isolation and culture of peripheral blood mononuclear cells eate the surface structure(s) present on the effector cells, the Peripheral blood mononuclear cells (PMNC) were isolated latter were first treated with different amounts Of pronase by standard Ficoll-Hypaque density gradient centrifugation. and neuraminidase. The effect of the enzymes on CYtOlYtic After several washes PMNC were cultured in 24-well plates activity against fresh melanoma cells was evaluated and com-at a concentration 106 cells/ml in R~~~ l a ~ containing pared with the NK-like activity against K562 target cells of the same effector population. A t a pronase concentration of lo% Grand Island, NY) in 0.01 mglml, no inhibition of NK-like activity was detected, the Presence of 200 U/ml recombinant IL-2 (rIL-2) (kindly whereas LAK activity was inhibited by more than 75%. In Provided by Biogen, Geneva, Switzerland). Cultures were addition, neuraminidase had no effect on NK-like activity, continued for 4 days.
even at I Ulml. whereas as little as 0.03 Ulml inhibited LAK activity by more than 75%. Metabolic inhibition of N-linked glycosylation with Tunicamycin prevented the generation of LAK activity, even when added late (18 hr before termination 1640 and resuspended at a concentration of 107/ml in medium of the culture). Tunicamycln, on the other hand, had no effect alone (control) or in the Same medium containing different ulations, in the presence of adherent cells, we analyzed the inhibitory activity of monoclonal antibodies (MAbs) to TI I, ~3 aminidase, Behringwerke, Marburg, FRG) (0.01, 0.03, 0.1, and T8 molecules. While all these MAbs strongly inhibited the 0.37 Or 1Ufml). The cells were incubated for 30 min at 37°C. specific target cell lysis by alloreactive CTLs, they had no At the end of the incubation period, FCS was added (1/10 of effect on the LAK activity.