CD2-mediated IL-12–dependent signals render human γδ-T
cells resistant to mitogen-induced apoptosis, permitting
the large-scale ex vivo expansion of functionally distinct
lymphocytes: implications for the development of adoptive
immunotherapy strategies
by Richard D. Lopez, Shan Xu, Ben Guo, Robert S. Negrin, and Edmund K. Waller
Blood
Volume 96(12):3827-3837
December 1, 2000
©2000 by American Society of Hematology
Anti-CD2 mAb S5.2 induces γδ–T-cell expansion from mitogen-stimulated PBMC
cultures.Cultures of human PBMC were initiated (day 0) by pre-incubating with IFN-γ (1000
U/mL) and then 24 hours later (day 1) with the addition of both OKT3 (10 ng/mL) and IL-2 (30...
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology
Expansion of γδ-T cell induced by anti-CD2 mAb S5.2 requires the presence of IL-12 and occurs
as a consequence of an increase in γδ–T-cell absolute numbers.Mitogen-stimulated PBMC
cultures were initiated as described above.
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology
Anti-CD2 mAb S5.2 induces γδ–T-cell expansion through an agonistic and not a blocking
interaction with CD2.(A) Anti-CD2 mAb S5.2 does not induce γδ–T-cell expansion by disrupting a
CD2–CD58 interaction.
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology
CD2-mediated IL-12–dependent signals render human γδ-T cells resistant to mitogen-induced
apoptosis: analysis by 4-color flow cytometry.PBMC cultures were initiated as described above.
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology
Both CD2-mediated signals and IL-12 signaling contribute to the expansion of apoptosisresistant γδ-T cells.On day 0, separate PBMC cultures were initiated.
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology
[3H]-thymidine incorporation in sorted, highly purified αβ- and γδ-T cells: late, but not early,
enhanced γδ–T-cell proliferation induced by mAb S5.2.Protected PBMC cultures were initiated
as described with all cultures receiving IFN-γ, IL-12, and mAb S5.2 ...
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology
IL-2 is a potent inducer of apoptosis in unprotected but not protected γδ-T cells.Unprotected and
protected PBMC cultures were initiated on day 0, as described above.
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology
Antitumor activity of apoptosis-resistant γδ-T cells demonstrated against human tumor cell
lines.Purified γδ- and αβ-T cells used as effector cells were sorted from 21-day cultures and
were routinely enriched from cultures to 97% or greater pure and 98% or ...
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology
Co-culture of tumor cells with apoptosis-resistant γδ-T cells: detection of tumor cell death.(A) T
lymphocytes alone (left plot) and HeLa target cells alone (center plot) have characteristic light
scatter properties that allow each cell population to be dis...
Richard D. Lopez et al. Blood 2000;96:3827-3837
©2000 by American Society of Hematology