Trial Information

Immunomonitoring of Children With Neuroblastoma for the Development of Antitumor Immunotherapy Strategies

The main objective of the study is the description of immune effectors in the blood, marrow
and tumor diagnosis.

During 3 years, this trial will include 30 children from pediatric oncology units of Lyon,
Saint-Etienne, Grenoble and Clermont-Ferrand.

The study duration is 5 years. Children follow-up scheduled for at least 2 years in order to
determine predictive factors of therapeutic efficiency and survival.

Multi-parametric marker sets (6-8 markers per sample) have already developed and validated
for analyzing the absolute amount and proportion of immune subpopulations (B, TCD4+, TCD8+,
Treg, NK, DC) and activation status (PD1, ICOS, CD39, CD73, CD62L, CCR7, CD45RO, CD45RA,
CD86, Ox40, CD137, CTLA4) on a small volume. At least, these analyses will be performed on
each blood and marrow sample. If the amount of blood and mononuclear cells harvested allows
it, functional analyses will be undertaken (intracytoplasmic cytokines in response to
activation for T, DC, NK; protocols been set up).

Immunostainings will be performed on tumor samples at diagnosis and after resection of the
primary tumor, in order to determine the expression and evolution of several
immunomodulatory molecules on neuroblastoma cells (HLA class I & II, HLA-G, IDO, IL10,…).,
and also determine the immune infiltrate within the tumor microenvironment (lymphocytes
Treg, cellules dendritiques, MDSC,…). The techniques used will mostly be those of classical
immunology (IHC, IF, FACS), and have already been set-up in our INSERM team for adult
tumors.

Children's plasma will be screened for specific anti-tumor immunoglobulins at diagnosis and
at key treatment time points. In the meanwhile, levels of circulating cytokines
concentrations will be evaluated by Luminex, especially those known to have inhibitory
effects on immune effectors: IL-4, IL-5, IL-6, IL-10, TGF-beta, HLA-Gs, TNF-alpha, IFN-gama,
IL-2, IL-12, IL-27, IL-17 and CD40L (already in place).