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Am J Transl Res 2012;4(1):114-126
Original Article
Implementing preclinical study findings to protocol design:
translational studies with alloreactive CTL for gliomas
Michelle J Hickey, Colin C Malone, Kate E Erickson, German G Gomez, Linda M Liau, Robert M Prins, Carol A
Kruse
Departments of Neurosurgery,1 David Geffen School of Medicine, Brain Research Institute,2 and the Jonsson
Comprehensive Cancer Center,3 University of California, Los Angeles, CA, USA.
Received December 31, 2011; accepted January, 2012; Epub January, 2012; Published January 15, 2012
Abstract: We are accruing patients to a Phase I dose escalation cellular therapy trial (www.clinicaltrials.gov,
NCT01144247) involving intratumoral placement of alloreactive cytotoxic T lymphocytes (alloCTL) for recurrent
gliomas. The trial is being conducted to confirm the findings of a prior pilot study that indicated this adjuvant
therapy may be beneficial in extending survival of recurrent WHO grade III gliomas. To reduce costs of the cellular
therapy, we tested a number of synthetic tissue culture media and found the AIM-V growth medium superior for
their growth. We also moved the production of the alloCTL from artificial capillary systems to less expensive
tissue culture bags. To standardize alloCTL infusates used for therapy, release criteria of ≥60% CD3+ and ≥60%
viability were established that consistently translated to a 4 hr cytotoxicity of ≥30% at a 30:1 effector to target ratio.
To allow time for completion of quality control testing and transport to the infusion site, we determined that 30,000
IU of human recombinant Interleukin-2 in the cellular infusates sufficiently retained cell viability and cytotoxicity to
allow a 10 hr expiration time to be placed on the infusates. We identified a cytotoxic T cell subset,
CD3+/CD8+/CD69+, that demonstrated upregulated IFN-γ production upon exposure to relevant target cells. The
phenotypic identification of this T cell subset was indicative of robust in vitro cytotoxic function and thus will be
followed to determine if it correlates with patient immune response to treatment. Finally, other therapeutic agents
routinely used for glioma treatment were integrated into an analysis of alloCTL cytotoxic functionality.
Temozolomide and bevacizumab do not adversely affect cytotoxic function of the alloCTL in the short-term, thus
providing rationale for further investigating combinatorial chemo-immunotherapy for gliomas. (AJTR1112006).
Keywords: Astrocytoma, glioma, CTL, temozolomide, bevacizumab, adoptive immunotherapy, cellular therapy,
immunotherapy
Full Text PDF
Address all correspondence to:
Carol A Kruse, PhD
UCLA David Geffen School of Medicine
Department of Neurosurgery
Gonda Center, Box 951761, MC 176122
695 Charles E Young Drive South
University of California Los Angeles
Los Angeles, CA 90095
Ph: 310-267-2535
email: ckruse@mednet.ucla.edu
Original Article
Implementing preclinical study findings to protocol design:
translational studies with alloreactive CTL for gliomas
Michelle J Hickey, Colin C Malone, Kate E Erickson, German G Gomez, Linda M Liau, Robert M Prins, Carol A
Kruse
Departments of Neurosurgery,1 David Geffen School of Medicine, Brain Research Institute,2 and the Jonsson
Comprehensive Cancer Center,3 University of California, Los Angeles, CA, USA.
Received December 31, 2011; accepted January, 2012; Epub January, 2012; Published January 15, 2012
Abstract: We are accruing patients to a Phase I dose escalation cellular therapy trial (www.clinicaltrials.gov,
NCT01144247) involving intratumoral placement of alloreactive cytotoxic T lymphocytes (alloCTL) for recurrent
gliomas. The trial is being conducted to confirm the findings of a prior pilot study that indicated this adjuvant
therapy may be beneficial in extending survival of recurrent WHO grade III gliomas. To reduce costs of the cellular
therapy, we tested a number of synthetic tissue culture media and found the AIM-V growth medium superior for
their growth. We also moved the production of the alloCTL from artificial capillary systems to less expensive
tissue culture bags. To standardize alloCTL infusates used for therapy, release criteria of ≥60% CD3+ and ≥60%
viability were established that consistently translated to a 4 hr cytotoxicity of ≥30% at a 30:1 effector to target ratio.
To allow time for completion of quality control testing and transport to the infusion site, we determined that 30,000
IU of human recombinant Interleukin-2 in the cellular infusates sufficiently retained cell viability and cytotoxicity to
allow a 10 hr expiration time to be placed on the infusates. We identified a cytotoxic T cell subset,
CD3+/CD8+/CD69+, that demonstrated upregulated IFN-γ production upon exposure to relevant target cells. The
phenotypic identification of this T cell subset was indicative of robust in vitro cytotoxic function and thus will be
followed to determine if it correlates with patient immune response to treatment. Finally, other therapeutic agents
routinely used for glioma treatment were integrated into an analysis of alloCTL cytotoxic functionality.
Temozolomide and bevacizumab do not adversely affect cytotoxic function of the alloCTL in the short-term, thus
providing rationale for further investigating combinatorial chemo-immunotherapy for gliomas. (AJTR1112006).
Keywords: Astrocytoma, glioma, CTL, temozolomide, bevacizumab, adoptive immunotherapy, cellular therapy,
immunotherapy
Full Text PDF
Address all correspondence to:
Carol A Kruse, PhD
UCLA David Geffen School of Medicine
Department of Neurosurgery
Gonda Center, Box 951761, MC 176122
695 Charles E Young Drive South
University of California Los Angeles
Los Angeles, CA 90095
Ph: 310-267-2535
email: ckruse@mednet.ucla.edu
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