TY - JOUR
T1 - Quantitative real-time RT-PCR as a method for monitoring T lymphocyte reactivity to full-length tyrosinase protein in vaccinated melanoma patients
AU - Housseau, Franck
AU - Lindsey, Kimberly R.
AU - Oberholtzer, Samuel D.
AU - Gonzales, Monica I.
AU - Boutin, Paula
AU - Moorthy, Anitha K.
AU - Shankara, Srinivas
AU - Roberts, Bruce L.
AU - Topalian, Suzanne L.
PY - 2002/8/1
Y1 - 2002/8/1
N2 - The major goal of therapeutic cancer vaccine trials is to mediate tumor regression. However, it is critically important to devise in vitro immunological assays that correlate with clinical outcome, for use as surrogate markers of vaccine efficacy. To date, clinical emphasis has been placed on peptide vaccines, but trends towards the use of more complex immunogens such as whole proteins require the development of efficient and sensitive methods for monitoring their immunological effects. In the context of a vaccination trial using full-length tyrosinase (Ty) to immunize patients with metastatic melanoma, a monitoring technique was developed in which autologous dendritic cells (DC) infected with a recombinant adenovirus encoding the Ty protein were used to assess the Ty-specific reactivity of fresh peripheral blood lymphocytes (PBL) collected from patients at different intervals during therapy. Quantitative real-time RT-PCR (qRT-PCR) was used to measure the production of cytokine mRNA by T cells following a 2.5-h incubation with Ty-expressing DC. Two out of ten patients studied demonstrated Ty protein-specific reactivity that increased during and after the period of vaccination. While one of these patients also reacted to an HLA-A1-compatible Ty peptide, the second did not recognize any of the known Ty epitopes, highlighting the importance of this technique for monitoring the effects of complex vaccines.
AB - The major goal of therapeutic cancer vaccine trials is to mediate tumor regression. However, it is critically important to devise in vitro immunological assays that correlate with clinical outcome, for use as surrogate markers of vaccine efficacy. To date, clinical emphasis has been placed on peptide vaccines, but trends towards the use of more complex immunogens such as whole proteins require the development of efficient and sensitive methods for monitoring their immunological effects. In the context of a vaccination trial using full-length tyrosinase (Ty) to immunize patients with metastatic melanoma, a monitoring technique was developed in which autologous dendritic cells (DC) infected with a recombinant adenovirus encoding the Ty protein were used to assess the Ty-specific reactivity of fresh peripheral blood lymphocytes (PBL) collected from patients at different intervals during therapy. Quantitative real-time RT-PCR (qRT-PCR) was used to measure the production of cytokine mRNA by T cells following a 2.5-h incubation with Ty-expressing DC. Two out of ten patients studied demonstrated Ty protein-specific reactivity that increased during and after the period of vaccination. While one of these patients also reacted to an HLA-A1-compatible Ty peptide, the second did not recognize any of the known Ty epitopes, highlighting the importance of this technique for monitoring the effects of complex vaccines.
KW - Adenovirus
KW - Dendritic cells
KW - Immune monitoring
KW - Quantitative real-time RT-PCR
KW - Tyrosinase
UR - http://www.scopus.com/inward/record.url?scp=0036679624&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036679624&partnerID=8YFLogxK
U2 - 10.1016/S0022-1759(02)00104-7
DO - 10.1016/S0022-1759(02)00104-7
M3 - Article
C2 - 12133625
AN - SCOPUS:0036679624
SN - 0022-1759
VL - 266
SP - 87
EP - 103
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -