Pediatric Surgery: Immunotherapy in Pediatric Solid Tumors

Dendritic Cell Development Dendritic Cell Vaccine Preparation

James D. Geiger

Assistant Professor of Surgery,
Section of Pediatric Surgery
University of Michigan Medical Center

Investigators

James D. Geiger, M.D., James J. Mule, Ph.D.

Techniques

  • Immunofluorescence staining
  • ELISA assay
  • Lymphocyte proliferation assay
  • Lymphocyte cytotoxicity assay
  • T-cell stimulation and culture
  • preparation of single cells from solid tumors
  • preparation of dendritic cells
  • isolation of CD-34 cells using magnetic micro beads (MACS)

Equipment

  • Co2 incubators
  • laminar airflow hoods
  • Inverted microscopes
  • light microscopes
  • table top centrifuge
  • floor centrifuge
  • cytospin II
  • MACS cell separator
  • FACScan flow cytometer
  • Revco - 80 freezer
  • Liquid nitrogen storage

Dendritic Cell Vaccines in The Treatment of Pediatric Solid Tumors

Collaborators: James J. Mule, Ph.D., Raymond Hutchinson, M.D.
Funding: NIH

The aims of this project are to investigate the use of dendritic cells as potent antigen presenting cells in a tumor vaccine to treat neuroblastoma and childhood sarcomas that have failed standard therapy. This therapy will be compared to vaccines consisting of tumor pulsed dendritic cells combined with chemokine secreting fibroblasts. Chemokines are essential for leukocyte function and trafficking and also enhance dendritic cell function. Pre-clinical studies will be performed to evaluate the capacity of dendritic cells with and without chemokines to detect T-cell specific responses to autologous neuroblastoma and sarcoma.

Other Studies

Other investigations will involve evaluating the use of CD 4( T- helper ) cells in adoptive immunotherapy. Optimal methods for expanding CD 4 cells and maintaining antigen specificity are being explored.

The laboratory will continue to explore the best method to generate dendritic cells from peripheral blood and stem cells (CD 34). Chemokines and cytokines are being used to optimize the function of dendritic cells in both antigen processing and presentation. In addition, we are actively trying to identify tumor specific antigens in pediatric solid tumors that might be utilized as targets for immunotherapy.