Biochemical characterization of antigen-specific glycosylation-inhibiting factor from antigen-specific suppressor T cells: II. The 55-kDa glycosylation-inhibiting factor peptide is a derivative of TCR α-chain and a subunit of antigen-specific glycosylation-inhibiting factor

Yasuyuki Ishii, Tatsumi Nakano, Kimishige Ishizaka

Research output: Contribution to journalArticle

Abstract

We isolated a unique TCR α-chain cDNA derived from the OVA-specific Ts hybridoma, 231F1. The TCR α-chain consists of Vα11.3, a unique Jα and a complete sequence of Cα region. Transfection of the TCR-α cDNA into a TCR-α-, TCR-β+ T cell line, 175.2, resulted in the expression of TCR-αβ, and the transfectant contained a 35-kDa peptide having the TCR-α-specific antigenic determinant. However, the stable transfectant failed to release a peptide with the TCR-α determinant upon stimulation with anti-CD3. In contrast, overexpression of the cDNA in the 231F1 cells markedly increased the formation of the 55-kDa peptide, which reacted with both anti-glycosylation-inhibiting factor (GIF) and the mAb H28-710. Definitive evidence for the relationship between the 55-kDa peptide and the TCR α-chain was obtained by transfection of the cDNA of the TCR α-chain with histidine tag into the 231F1 cells. The 55-kDa GIF peptide formed by stable tranfectants of the TCR-α-tag cDNA bound to Ni+-nitrilotriacetic acid-agarose. Upon stimulation with anti-CD3, a stable transfectant of the TCR-α cDNA formed OVA-specific GIF which contained the 55-kDa GIF peptide, and bound not only to anti-TCR-α column but also to anti-TCR-β column. The results indicate that the OVA-specific GIF consists of the TCR-α+ 55-kDa GIF and another peptide with TCR-β determinant. It was found that association of the TCR-β+ peptide with the 55-kDa GIF is required for binding of the factor to OVA, but not essential for the formation and release of the latter peptide.

Original languageEnglish (US)
Pages (from-to)1735-1742
Number of pages8
JournalJournal of Immunology
Volume156
Issue number5
StatePublished - Mar 1 1996
Externally publishedYes

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T-Lymphocytes
Antigens
Peptides
Complementary DNA
Transfection
Nitrilotriacetic Acid
immunoglobulin-binding factors
Hybridomas
Histidine
Sepharose
Epitopes
Cell Line

ASJC Scopus subject areas

  • Immunology

Cite this

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title = "Biochemical characterization of antigen-specific glycosylation-inhibiting factor from antigen-specific suppressor T cells: II. The 55-kDa glycosylation-inhibiting factor peptide is a derivative of TCR α-chain and a subunit of antigen-specific glycosylation-inhibiting factor",
abstract = "We isolated a unique TCR α-chain cDNA derived from the OVA-specific Ts hybridoma, 231F1. The TCR α-chain consists of Vα11.3, a unique Jα and a complete sequence of Cα region. Transfection of the TCR-α cDNA into a TCR-α-, TCR-β+ T cell line, 175.2, resulted in the expression of TCR-αβ, and the transfectant contained a 35-kDa peptide having the TCR-α-specific antigenic determinant. However, the stable transfectant failed to release a peptide with the TCR-α determinant upon stimulation with anti-CD3. In contrast, overexpression of the cDNA in the 231F1 cells markedly increased the formation of the 55-kDa peptide, which reacted with both anti-glycosylation-inhibiting factor (GIF) and the mAb H28-710. Definitive evidence for the relationship between the 55-kDa peptide and the TCR α-chain was obtained by transfection of the cDNA of the TCR α-chain with histidine tag into the 231F1 cells. The 55-kDa GIF peptide formed by stable tranfectants of the TCR-α-tag cDNA bound to Ni+-nitrilotriacetic acid-agarose. Upon stimulation with anti-CD3, a stable transfectant of the TCR-α cDNA formed OVA-specific GIF which contained the 55-kDa GIF peptide, and bound not only to anti-TCR-α column but also to anti-TCR-β column. The results indicate that the OVA-specific GIF consists of the TCR-α+ 55-kDa GIF and another peptide with TCR-β determinant. It was found that association of the TCR-β+ peptide with the 55-kDa GIF is required for binding of the factor to OVA, but not essential for the formation and release of the latter peptide.",
author = "Yasuyuki Ishii and Tatsumi Nakano and Kimishige Ishizaka",
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T1 - Biochemical characterization of antigen-specific glycosylation-inhibiting factor from antigen-specific suppressor T cells

T2 - II. The 55-kDa glycosylation-inhibiting factor peptide is a derivative of TCR α-chain and a subunit of antigen-specific glycosylation-inhibiting factor

AU - Ishii, Yasuyuki

AU - Nakano, Tatsumi

AU - Ishizaka, Kimishige

PY - 1996/3/1

Y1 - 1996/3/1

N2 - We isolated a unique TCR α-chain cDNA derived from the OVA-specific Ts hybridoma, 231F1. The TCR α-chain consists of Vα11.3, a unique Jα and a complete sequence of Cα region. Transfection of the TCR-α cDNA into a TCR-α-, TCR-β+ T cell line, 175.2, resulted in the expression of TCR-αβ, and the transfectant contained a 35-kDa peptide having the TCR-α-specific antigenic determinant. However, the stable transfectant failed to release a peptide with the TCR-α determinant upon stimulation with anti-CD3. In contrast, overexpression of the cDNA in the 231F1 cells markedly increased the formation of the 55-kDa peptide, which reacted with both anti-glycosylation-inhibiting factor (GIF) and the mAb H28-710. Definitive evidence for the relationship between the 55-kDa peptide and the TCR α-chain was obtained by transfection of the cDNA of the TCR α-chain with histidine tag into the 231F1 cells. The 55-kDa GIF peptide formed by stable tranfectants of the TCR-α-tag cDNA bound to Ni+-nitrilotriacetic acid-agarose. Upon stimulation with anti-CD3, a stable transfectant of the TCR-α cDNA formed OVA-specific GIF which contained the 55-kDa GIF peptide, and bound not only to anti-TCR-α column but also to anti-TCR-β column. The results indicate that the OVA-specific GIF consists of the TCR-α+ 55-kDa GIF and another peptide with TCR-β determinant. It was found that association of the TCR-β+ peptide with the 55-kDa GIF is required for binding of the factor to OVA, but not essential for the formation and release of the latter peptide.

AB - We isolated a unique TCR α-chain cDNA derived from the OVA-specific Ts hybridoma, 231F1. The TCR α-chain consists of Vα11.3, a unique Jα and a complete sequence of Cα region. Transfection of the TCR-α cDNA into a TCR-α-, TCR-β+ T cell line, 175.2, resulted in the expression of TCR-αβ, and the transfectant contained a 35-kDa peptide having the TCR-α-specific antigenic determinant. However, the stable transfectant failed to release a peptide with the TCR-α determinant upon stimulation with anti-CD3. In contrast, overexpression of the cDNA in the 231F1 cells markedly increased the formation of the 55-kDa peptide, which reacted with both anti-glycosylation-inhibiting factor (GIF) and the mAb H28-710. Definitive evidence for the relationship between the 55-kDa peptide and the TCR α-chain was obtained by transfection of the cDNA of the TCR α-chain with histidine tag into the 231F1 cells. The 55-kDa GIF peptide formed by stable tranfectants of the TCR-α-tag cDNA bound to Ni+-nitrilotriacetic acid-agarose. Upon stimulation with anti-CD3, a stable transfectant of the TCR-α cDNA formed OVA-specific GIF which contained the 55-kDa GIF peptide, and bound not only to anti-TCR-α column but also to anti-TCR-β column. The results indicate that the OVA-specific GIF consists of the TCR-α+ 55-kDa GIF and another peptide with TCR-β determinant. It was found that association of the TCR-β+ peptide with the 55-kDa GIF is required for binding of the factor to OVA, but not essential for the formation and release of the latter peptide.

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