PMID- 10838667
OWN - NLM
STAT- MEDLINE
DCOM- 20000816
LR  - 20191104
IS  - 1524-9557 (Print)
IS  - 1524-9557 (Linking)
VI  - 23
IP  - 3
DP  - 2000 May-Jun
TI  - Development of a polynucleotide vaccine from melanoma antigen recognized by T 
      cells-1 and recombinant protein from melanoma antigen recognized by T cells-1 for 
      melanoma vaccine clinical trials.
PG  - 379-86
AB  - MART-1, a melanoma antigen recognized by T cells-1, is a melanocyte 
      lineage-differentiation antigen expressed only in melanocytes and melanoma cells. 
      This protein is recognized by many T-lymphocyte lines that are human leukocyte 
      antigen (HLA)-A2 restricted and melanoma reactive. These observations have 
      culminated in an array of clinical trials of MART-1 immunization using 
      recombinant viruses or MART-1 immunodominant peptides. Polynucleotide 
      immunization is a promising alternative to recombinant viral vaccines that allows 
      delivery of the full-length cDNA encoding all potential peptide epitopes in a 
      vector that is uncompromised by anti-viral immunity. In preparation for a phase I 
      clinical trial of MART-1 polynucleotide immunization in patients with resected 
      melanoma who were at significant risk for recurrence, the authors constructed a 
      plasmid DNA encoding the MART-1 cDNA under transcriptional regulatory control of 
      the cytomegalovirus immediate early promoter-enhancer and partially deleted 
      intron A. This plasmid directs high-level MART-1 expression in transduced 
      myoblasts and maturing myocytes diffusely throughout the cytoplasm. Immunization 
      of mice with this construct by intramuscular injection elicited MART-1-specific 
      immune responses in all animals. Previous trials of MART-1 immunization have been 
      unable to examine the humoral immune response to MART-1 because of a lack of 
      sufficient, highly purified protein. We have produced and purified Escherichia 
      coli recombinant MART-1 protein using a glutathione-S-transferase fusion protein 
      expression system. Protein staining of a sodium dodecyl sulfate-polyacrylamide 
      gel electrophoresis revealed a band of MART-1 protein at approximately 20 kD; and 
      Western immunoblotting with an anti-MART-1 monoclonal antibody confirmed a 
      doublet at approximately 20 kD. These findings are consistent with previous 
      reports using different expression systems for recombinant MART-1. This protein 
      preparation functioned well in enzyme-linked immunosorbent assays (ELISAs) to 
      detect anti-MART-1 antibody responses in a mouse model; and a panel of healthy 
      donor human sera showed minimal binding to ELISA plates coated with the protein, 
      supporting its utility in monitoring human anti-MART-1 antibody responses. The 
      glutathione-S-transferase fusion method yielded approximately 200 micrograms 
      MART-1 per 2-L bacterial culture, enough to coat 100 ELISA plates.
FAU - Lee, S W
AU  - Lee SW
AD  - Department of Medicine, University of Alabama at Birmingham 35294-3300, USA.
FAU - Li, H
AU  - Li H
FAU - Strong, T V
AU  - Strong TV
FAU - Moore, S E
AU  - Moore SE
FAU - Conry, R M
AU  - Conry RM
LA  - eng
GR  - CA 61397/CA/NCI NIH HHS/United States
GR  - CA 68256/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Immunother
JT  - Journal of immunotherapy (Hagerstown, Md. : 1997)
JID - 9706083
RN  - 0 (Antigens, Neoplasm)
RN  - 0 (Cancer Vaccines)
RN  - 0 (MART-1 Antigen)
RN  - 0 (MLANA protein, human)
RN  - 0 (Mlana protein, mouse)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Vaccines, DNA)
SB  - IM
MH  - Animals
MH  - Antibody Formation
MH  - Antigens, Neoplasm/*immunology/*isolation & purification
MH  - Cancer Vaccines/*immunology
MH  - Cell Line
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Escherichia coli/genetics
MH  - Female
MH  - Immunohistochemistry
MH  - MART-1 Antigen
MH  - Melanoma/*immunology
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Neoplasm Proteins/genetics/*immunology
MH  - Plasmids
MH  - Recombinant Proteins/immunology/isolation & purification
MH  - Vaccines, DNA/*immunology
EDAT- 2000/06/06 09:00
MHDA- 2000/08/19 11:00
CRDT- 2000/06/06 09:00
PHST- 2000/06/06 09:00 [pubmed]
PHST- 2000/08/19 11:00 [medline]
PHST- 2000/06/06 09:00 [entrez]
AID - 10.1097/00002371-200005000-00011 [doi]
PST - ppublish
SO  - J Immunother. 2000 May-Jun;23(3):379-86. doi: 10.1097/00002371-200005000-00011.