Induction of anti-melanoma CTL response using DC transfected with mutated mRNA encoding full-length Melan-A/MART-1 antigen with an A27L amino acid substitution

Zeinab Abdel-Wahab, Matthew F. Kalady, Sirisha Emani, Mark W. Onaitis, Omar I. Abdel-Wahab, Robin Cisco, Lee Wheless, Tsung Yen Cheng, Douglas Tyler, Scott K. Pruitt

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Modification of the parental immunodominant Melan-A/MART-1 peptide (MART-126-35) by replacing the alanine with leucine (A27L) enhances its immunogenicity. Because of the reported advantages of RNA over peptides in DC vaccines, we sought to mutate the MART-1 gene to encode a full-length MART-1 antigen with an A27L amino acid substitution. Human DC were transfected with A27L-mutated MART-1 RNA (A27L RNA) or native MART-1 RNA, and then used to stimulate autologous T cells from a series of 8 HLA-A2+ volunteers. After three stimulations, all CTL induced with DC/A27L RNA exhibited more tetramer+ cells, and demonstrated stronger antigen-specific IFNγ-secreting activity compared to CTL induced with DC/native RNA. A potent MART-1-specific, and predominantly class-I-restricted lysis was detected in most CTL induced with DC/A27L RNA, while native RNA-induced CTL showed minimal and non-specific lysis. HLA-A2+ DC and MART-1 negative/A2+ melanoma cells transfected with the A27L RNA were recognized and killed by MART-1-specific CTL, suggesting that these APC efficiently processed the A27L RNA and presented correct MART-1-specific epitope(s). In summary, introducing an A27L mutation into the MART-1 full-length mRNA sequence enhanced the immunogenicity of the encoded MART-1 Ag. The ease with which such a mutation can be made in RNA presents another potential advantage of using RNA for immunotherapy. Our results support considering this strategy for enhancing the immunogenicity of DC-based RNA vaccines.

Original languageEnglish (US)
Pages (from-to)86-97
Number of pages12
JournalCellular Immunology
Volume224
Issue number2
DOIs
StatePublished - Jan 1 2003
Externally publishedYes

Fingerprint

MART-1 Antigen
Amino Acid Substitution
Melanoma
RNA
Messenger RNA
HLA-A2 Antigen
Vaccines
HLA-DQ Antigens
Peptides
Mutation
varespladib methyl
Leucine
Alanine
Immunotherapy

Keywords

  • A27L-mutated RNA
  • CTL
  • DC
  • Melan-A MART-1
  • Melanoma

ASJC Scopus subject areas

  • Immunology

Cite this

Induction of anti-melanoma CTL response using DC transfected with mutated mRNA encoding full-length Melan-A/MART-1 antigen with an A27L amino acid substitution. / Abdel-Wahab, Zeinab; Kalady, Matthew F.; Emani, Sirisha; Onaitis, Mark W.; Abdel-Wahab, Omar I.; Cisco, Robin; Wheless, Lee; Cheng, Tsung Yen; Tyler, Douglas; Pruitt, Scott K.

In: Cellular Immunology, Vol. 224, No. 2, 01.01.2003, p. 86-97.

Research output: Contribution to journalArticle

Abdel-Wahab, Zeinab ; Kalady, Matthew F. ; Emani, Sirisha ; Onaitis, Mark W. ; Abdel-Wahab, Omar I. ; Cisco, Robin ; Wheless, Lee ; Cheng, Tsung Yen ; Tyler, Douglas ; Pruitt, Scott K. / Induction of anti-melanoma CTL response using DC transfected with mutated mRNA encoding full-length Melan-A/MART-1 antigen with an A27L amino acid substitution. In: Cellular Immunology. 2003 ; Vol. 224, No. 2. pp. 86-97.
@article{a25af769299f438eba445a2b605a2191,
title = "Induction of anti-melanoma CTL response using DC transfected with mutated mRNA encoding full-length Melan-A/MART-1 antigen with an A27L amino acid substitution",
abstract = "Modification of the parental immunodominant Melan-A/MART-1 peptide (MART-126-35) by replacing the alanine with leucine (A27L) enhances its immunogenicity. Because of the reported advantages of RNA over peptides in DC vaccines, we sought to mutate the MART-1 gene to encode a full-length MART-1 antigen with an A27L amino acid substitution. Human DC were transfected with A27L-mutated MART-1 RNA (A27L RNA) or native MART-1 RNA, and then used to stimulate autologous T cells from a series of 8 HLA-A2+ volunteers. After three stimulations, all CTL induced with DC/A27L RNA exhibited more tetramer+ cells, and demonstrated stronger antigen-specific IFNγ-secreting activity compared to CTL induced with DC/native RNA. A potent MART-1-specific, and predominantly class-I-restricted lysis was detected in most CTL induced with DC/A27L RNA, while native RNA-induced CTL showed minimal and non-specific lysis. HLA-A2+ DC and MART-1 negative/A2+ melanoma cells transfected with the A27L RNA were recognized and killed by MART-1-specific CTL, suggesting that these APC efficiently processed the A27L RNA and presented correct MART-1-specific epitope(s). In summary, introducing an A27L mutation into the MART-1 full-length mRNA sequence enhanced the immunogenicity of the encoded MART-1 Ag. The ease with which such a mutation can be made in RNA presents another potential advantage of using RNA for immunotherapy. Our results support considering this strategy for enhancing the immunogenicity of DC-based RNA vaccines.",
keywords = "A27L-mutated RNA, CTL, DC, Melan-A MART-1, Melanoma",
author = "Zeinab Abdel-Wahab and Kalady, {Matthew F.} and Sirisha Emani and Onaitis, {Mark W.} and Abdel-Wahab, {Omar I.} and Robin Cisco and Lee Wheless and Cheng, {Tsung Yen} and Douglas Tyler and Pruitt, {Scott K.}",
year = "2003",
month = "1",
day = "1",
doi = "10.1016/j.cellimm.2003.08.005",
language = "English (US)",
volume = "224",
pages = "86--97",
journal = "Cellular Immunology",
issn = "0008-8749",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Induction of anti-melanoma CTL response using DC transfected with mutated mRNA encoding full-length Melan-A/MART-1 antigen with an A27L amino acid substitution

AU - Abdel-Wahab, Zeinab

AU - Kalady, Matthew F.

AU - Emani, Sirisha

AU - Onaitis, Mark W.

AU - Abdel-Wahab, Omar I.

AU - Cisco, Robin

AU - Wheless, Lee

AU - Cheng, Tsung Yen

AU - Tyler, Douglas

AU - Pruitt, Scott K.

PY - 2003/1/1

Y1 - 2003/1/1

N2 - Modification of the parental immunodominant Melan-A/MART-1 peptide (MART-126-35) by replacing the alanine with leucine (A27L) enhances its immunogenicity. Because of the reported advantages of RNA over peptides in DC vaccines, we sought to mutate the MART-1 gene to encode a full-length MART-1 antigen with an A27L amino acid substitution. Human DC were transfected with A27L-mutated MART-1 RNA (A27L RNA) or native MART-1 RNA, and then used to stimulate autologous T cells from a series of 8 HLA-A2+ volunteers. After three stimulations, all CTL induced with DC/A27L RNA exhibited more tetramer+ cells, and demonstrated stronger antigen-specific IFNγ-secreting activity compared to CTL induced with DC/native RNA. A potent MART-1-specific, and predominantly class-I-restricted lysis was detected in most CTL induced with DC/A27L RNA, while native RNA-induced CTL showed minimal and non-specific lysis. HLA-A2+ DC and MART-1 negative/A2+ melanoma cells transfected with the A27L RNA were recognized and killed by MART-1-specific CTL, suggesting that these APC efficiently processed the A27L RNA and presented correct MART-1-specific epitope(s). In summary, introducing an A27L mutation into the MART-1 full-length mRNA sequence enhanced the immunogenicity of the encoded MART-1 Ag. The ease with which such a mutation can be made in RNA presents another potential advantage of using RNA for immunotherapy. Our results support considering this strategy for enhancing the immunogenicity of DC-based RNA vaccines.

AB - Modification of the parental immunodominant Melan-A/MART-1 peptide (MART-126-35) by replacing the alanine with leucine (A27L) enhances its immunogenicity. Because of the reported advantages of RNA over peptides in DC vaccines, we sought to mutate the MART-1 gene to encode a full-length MART-1 antigen with an A27L amino acid substitution. Human DC were transfected with A27L-mutated MART-1 RNA (A27L RNA) or native MART-1 RNA, and then used to stimulate autologous T cells from a series of 8 HLA-A2+ volunteers. After three stimulations, all CTL induced with DC/A27L RNA exhibited more tetramer+ cells, and demonstrated stronger antigen-specific IFNγ-secreting activity compared to CTL induced with DC/native RNA. A potent MART-1-specific, and predominantly class-I-restricted lysis was detected in most CTL induced with DC/A27L RNA, while native RNA-induced CTL showed minimal and non-specific lysis. HLA-A2+ DC and MART-1 negative/A2+ melanoma cells transfected with the A27L RNA were recognized and killed by MART-1-specific CTL, suggesting that these APC efficiently processed the A27L RNA and presented correct MART-1-specific epitope(s). In summary, introducing an A27L mutation into the MART-1 full-length mRNA sequence enhanced the immunogenicity of the encoded MART-1 Ag. The ease with which such a mutation can be made in RNA presents another potential advantage of using RNA for immunotherapy. Our results support considering this strategy for enhancing the immunogenicity of DC-based RNA vaccines.

KW - A27L-mutated RNA

KW - CTL

KW - DC

KW - Melan-A MART-1

KW - Melanoma

UR - http://www.scopus.com/inward/record.url?scp=85058201519&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85058201519&partnerID=8YFLogxK

U2 - 10.1016/j.cellimm.2003.08.005

DO - 10.1016/j.cellimm.2003.08.005

M3 - Article

VL - 224

SP - 86

EP - 97

JO - Cellular Immunology

JF - Cellular Immunology

SN - 0008-8749

IS - 2

ER -