Vol. 7, 917s-924s, March 2001 (Suppl.) Clinical Cancer Research 917s Intrathecal Cytotoxic T-Cell Immunotherapy for Metastatic Leptomeningeal Melanoma a Annette R. Clemons-Miller, Gurkamal S. Chatta, Laura Hutchins, Edgardo J. Angtuaco, Antonella Ravaggi, Alessandro D. Santin, and Martin J. Cannon 2 Departments of Geriatrics [A. R. C-M., G. S. C.], Medicine [G. S. C., L. H.], Radiology [E. J. A.], Obstetrics and Gynecology [A. R., A. D. S.], and Microbiology and Immunology [A. R. C-M., M. J. C.], University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, and Division of Gynecologic Oncology, University of Brescia Medical School, Brescia, Italy [A. R., A. D. S.] Abstract A 49-year-old patient with primary, recurrent melanoma on the lower extremity developed metastatic leptomeningeal melanoma that did not respond to treatment with radiation therapy or intrathecal interleukin 2 (IL-2). Disease was char- acterized by neurological symptoms, including loss of hearing, loss of short-term memory, and gait disturbance. CD8 + CTLs were generated in vitro using autologous dendritic cells pulsed with peptides from the melanoma-associated antigens tyrosin- ase (145-156), Melan-A/MART-1 (26-35), and gpl00/Pmel 17 (209-217). The CTLs exhibited up to 74% specific lysis against peptide-pulsed autologous EBV-transformed B cells, with Melan-A-specific CTLs yielding the greatest lytic activity. CD8 + CTLs possessed a type 1 cytokine profile, expressing tumor necrosis factor oL and IFN~/but not IL-4. Infusions of CTLs were supported with systemic low-dose IL-2 administra- tion. 111In labeling and computerized gamma imaging were used to monitor the distribution of CTLs up to 48 h after infusion. Intra-arterial delivery via the right carotid artery was followed by redistribution of the CTLs to the lungs, liver, and spleen within 16 h. In contrast, delivery via an indwelling Ommaya reservoir resulted in prolonged retention of CTLs within the brain for at least 48 h after infusion. Marked but transient elevations in tumor necrosis factor e~, IFN-% and IL-6 in the cerebrospinal fluid were observed within 4 h of CTL infusion. There was no evidence of tumor progression throughout the treatment period, and clinically the patient showed some resolution of neurological symptoms. Introduction Although the immunogenic potential of melanoma has been recognized for some time, clinical trials involving the use of tumor-infiltrating lymphocytes or lymphokine-activated killer cells in the treatment of malignant melanoma have met with only modest success (1-6). The reasons for the limited efficacy may include poor definition of the effector cells, failure of the effector cells to recognize defined antigens, and poor localization of the effector cells to the site of the tumor. The recent identification of defined melanoma-associated tumor an- tigens such as gp 100, Melan-A/MART-1, tyrosinase, MAGE-1, and MAGE-3 (7-12), combined with the ability to induce T-cell responses of defined function and phenotype, has led to a strong resurgence of interest in immunotherapy of melanoma. Most notably, the application of DCs 3 as powerful inducers of mela- noma tumor antigen-specific CD8 + cytotoxic CTL responses has provided investigators with the tools necessary for a targeted approach to the treatment of disease (13-19). Notwithstanding the major advances in DC-based induc- tion of tumor-antigen-specific T-cell responses in patients with malignant disease, clinical administration of autologous anti- gen-specific CTLs presents some practical challenges. One of the more important issues centers on election of an appropriate route of delivery for optimal localization and retention of CTLs at the tumor site. The use of radioisotopic cell labeling and subsequent imaging of T-cell localization in the patient affords us the opportunity to tailor the route and method of CTL immunotherapy for optimum delivery to the tumor. Isotopes such as ~In can be used for short-term visualization and tracking of cellular components in real time using standard scanning techniques (20-23). In this study, we have used autologous DCs pulsed with defined HLA-restricted peptides from the gpl00, Melan-A/ MART-l, and tyrosinase melanoma antigens to stimulate CD8 + CTLs for immunotherapeutic treatment of a patient with metastatic leptomeningeal melanoma. Antigen-specific CTLs were delivered through catheterization of the internal carotid artery or local deliv- ery through an indwelling intracranial Ommaya reservoir. ~11In labeling and computerized gamma imaging were used to monitor CTL migration and elucidation of the most effective route of delivery. CTL activity in vivo was assessed indirectly through measurement of T-cell and inflammatory cytokine production in the CSF at various time points after treatment. Materials and Methods Case History. A 49-year-old woman presented with a melanocytic nevus (Breslow thickness, 0.8 ram), which was re- moved in February 1995. A local recurrence (Breslow thickness, 1 This study was supported by NIH Grants CA 63931 (to M. J. C.) and CA 76033 (G. S. C.) and the Arkansas Cancer Research Center. a To whom requests for reprints should be addressed, at Department of Microbiology and Immunology, Mail Slot 511, University of Arkansas for Medical Sciences, 4301 West Markham, Little Rock, AR 72205. Phone: (501)296-1254; Fax: (501)686-5359; E-mail: cannonmartin@ exchange.uams.edu. 3 The abbreviations used are: DC, dendritic cell; PBMC, peripheral blood mononuclear cell; CSF, cerebrospinal fluid; MRI, magnetic res- onance imaging; GM-CSF, granulocyte/macrophage-colony stimulating factor; TNF, tumor necrosis factor; IL, interleukin; NK, natural killer; PMA, phorbol myristate acetate; CNS, central nervous system. Research. on October 21, 2021. © 2001 American Association for Cancer clincancerres.aacrjournals.org Downloaded from