Potential use of lymph node-derived HPV-specific T cells for adoptive cell therapy of cervical cancer.
Adoptive transfer of tumor-specific T cells, expanded from tumor-infiltrating lymphocytes or from peripheral blood, is a promising immunotherapeutic approach for the treatment of cancer. Here, we studied whether the tumor-draining lymph nodes (TDLN) of patients with human papillomavirus (HPV)-induced cervical cancer can be used as a source for ACT. The objectives were to isolate lymph node mononuclear cells (LNMC) from TDLN and optimally expand HPV-specific CD4+ and CD8+ T cells under clinical grade conditions. TDLN were isolated from 11 patients with early-stage cervical cancer during radical surgery. Isolated lymphocytes were expanded in the presence of HPV16 E6 and E7 clinical grade synthetic long peptides and IL-2 for 22 days and then analyzed for HPV16 specificity by proliferation assay, multiparameter flow cytometry and cytokine analysis as well as for CD25 and FoxP3 expression. Stimulation of LNMC resulted in expansion of polyclonal HPV-specific T cells in all patients. On average a 36-fold expansion of a CD4+ and/or CD8+ HPV16-specific T cell population was observed, which maintained its capacity for secondary expansion. The T helper type 1 cytokine IFNγ was produced in all cell cultures and in some cases also the Th2 cytokines IL-10 and IL-5. The procedure was highly reproducible, as evidenced by complete repeats of the stimulation procedures under research and under full good manufacturing practice conditions. In conclusion, TDLN represent a rich source of polyclonal HPV16 E6- and E7-specific T cells, which can be expanded under clinical grade conditions for adoptive immunotherapy in patients with cervical cancer.
Department of Gynecology, Leiden University Medical Center, Leiden, The Netherlands.
Mariëtte I E
0000-0001-5432-5001
MI
van Poelgeest
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands.|Department of Microbiology Diagnostic, University of Rome "La Sapienza", Rome, Italy.
Valeria V
VV
Visconti
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands.
Zohara
Z
Aghai
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands.
Vanessa J
VJ
van Ham
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands.|Department of Medical Microbiology, Free University Medical Center, Amsterdam, The Netherlands.
Moniek
M
Heusinkveld
Clinical Pharmacy and Toxicology, Leiden University Medical Center, Leiden, The Netherlands.
Maarten L
ML
Zandvliet
Clinical Pharmacy and Toxicology, Leiden University Medical Center, Leiden, The Netherlands.
A Rob P M
AR
Valentijn
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands.
Renske
R
Goedemans
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands.
Caroline E
CE
van der Minne
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands.
Els M E
EM
Verdegaal
Department of Gynecology, Leiden University Medical Center, Leiden, The Netherlands.
J Baptist M Z
JB
Trimbos
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands.
Sjoerd H
SH
van der Burg
Department of Medical Oncology, Leiden University Medical Center, Building 1, K1-P, PO box 9600, 2300 RC, Leiden, The Netherlands. m.j.p.schoenmaekers-welters@lumc.nl.
Marij J P
MJ
Welters
D000328:Adult; D000368:Aged; D005260:Female; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008198:Lymph Nodes / Q000276:immunology* / Q000473:pathology; D008875:Middle Aged; D013601:T-Lymphocytes / Q000276:immunology*; D002583:Uterine Cervical Neoplasms / Q000276:immunology* / Q000628:therapy
Adoptive cell transfer; Cervical cancer; Good manufacturing practice; HPV-specific T cells; Human papillomavirus; Tumor-draining lymph node