• Vaccine Name: Cancer Retroviral Vector Vaccine encoding Cd40lg
• Target Pathogen: Cancer
• Target Disease: Cancer
• Vaccine Ontology ID: VO_0011372
• Type: Recombinant vector vaccine
• Status: Research
• Antigen: CD40 Ligand
• Cd40lg gene engineering:
  • Type: Recombinant vector construction
  • Description: Murine CD40 ligand (mCD40L) gene was cloned from a c D N A library generated from activated T cells (Coleclough, 1993) using seminested P C R primers that incorporated a Kpn I site at the 5' end of the gene and a Cta I site at the 3' end of the gene using AmpliTaq (Perkin-Elmer, Branchburg, NJ) according to the manufacturer's protocols. The polymerase chain reaction (PCR) product was inserted into p G E M 7 Z (Pharmacia Biotech Inc., Piscataway, NJ) and the correct D N A sequence of a full-length clone as compared to published data (Armitage et al, 1992) was confirmed. The m u C D 4 0 L gene was then subcloned into the Gla retroviral vector derived from Moloney murine leukemia virus (Genetic Therapy, Inc., Gaithersburg, M D ) using the Xho I and Hind II sites (pGla.mCD40L) and used to manufacture a producer cell line (Grossmann et al., 1997).
  • Detailed Gene Information: Click Here.
• Immunization Route: subcutaneous injection

Mouse Response

• Host Strain: A/J
• Vaccination Protocol: A/J (H^'') female mice (Jackson Labs) received a single subcutaneous (s.c.) injection consisting of 2 X 10* cells total unless otherwise noted in the figure legends. The cells were either neuro-2a/neo cells ( 0 % C D 4 0 L positive), neuro-2a/CD40L (70% CD40L positive), or neuro-2a/neo cells mixed with various
  numbers of neuro-2a/CD40L cells to the appropriate percentages of CD40L-positive cells (Grossmann et al., 1997).
• Challenge Protocol: Antitumor effects were tested with subsequent challenge with parental neuro-2a cells (Grossmann et al., 1997).
• Efficacy: Transgenic expression of the CD40L (Cd40lg) increased immune responses against a weakly immunogenic murine tumor, neuro-2a. Tumor cells were transduced with a retroviral construct containing the CD40L gene and co-injected with variable numbers of non-CD40L transduced cells into syngeneic mice. Mice injected with cells that expressed CD40L had a significant reduction in average tumor size as compared to controls (p < 0.0001). In addition, survival of the neuro-2a/CD40L mice was 48 days versus 34 days for the neuro-2a/neo controls (p < 0.02). Expression of CD40L by less than 1.5% of neuro-2a cells was sufficient for significant antitumor effects (p < 0.001). These antitumor effects protected mice from subsequent challenge with parental neuro-2a cells (Grossmann et al., 1997).