A human acute leukemia-derived T-cell line produces two inhibitor factors which suppress lymphocyte proliferation: Characterization and purification of the molecules

P. G. Montaldo, M. Lanciotti, E. Castagnola, M. T. Parodi, C. Cirillo, P. Cornaglia-Ferraris, M. Ponzoni

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The crude supernatant of an CD1+/CD8+ T-cell clone (GI-CO-T-9) established from a long standing culture of an acute T-lymphoblastic leukemia (T-ALL) was shown to inhibit the responsiveness of normal PBMC to PHA. This clone does not produce TNF-α, TNF-β, α-IFN, τ-IFN, IL-1, IL-2 and has no NK-like activity. The crude supernatant has been subjected to a multi-step chromatographic fractioning. Preparative gel permeation HPLC allowed us to recover two peaks of biologic activity in the range of 100-120 kDa and 75-85 kDa (referred to as 'high molecular mass inhibitor factor', HMMIF, and 'low molecular mass inhibitor factor', LMMIF, respectively). Both fractions were then subjected to anion exchange HPLC: HMMIF was recovered in fractions eluting at 0.04 M NaCl while LMMIF eluted at higher ionic strength (0.48 M NaCl). The fractions with biologic activity recovered from anion exchange HPLC have been subjected to hydrophobic interaction HPLC (HIC) for final purification. The highly purified material was characterized by polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate (SDS) revealing single bands of 115 kDa and 80 kDa. The isoelectric points (pI), determined by flat-bed isoelectricfocusing, were 7.4 for HMMIF and 3.5-3.6 for LMMIF. Studies on temperature lability indicated that both proteins are stable for 3-4 hours at room temperature (RT), 24-36 hours at +4°C and 7-10 days at -80°C.

Original languageEnglish
Pages (from-to)413-427
Number of pages15
JournalLymphokine Research
Volume7
Issue number4
Publication statusPublished - 1988

Fingerprint

Leukemia
High Pressure Liquid Chromatography
Lymphocytes
Cell Line
Anions
Clone Cells
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
Temperature
Isoelectric Point
Carbon Monoxide
Interleukin-1
Hydrophobic and Hydrophilic Interactions
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Sodium Dodecyl Sulfate
Osmolar Concentration
Interleukin-2
Polyacrylamide Gel Electrophoresis
Gels
T-Lymphocytes
Proteins

ASJC Scopus subject areas

  • Immunology
  • Hematology

Cite this

@article{a3fc6f84a3d145b88cdddef4f3f759a9,
title = "A human acute leukemia-derived T-cell line produces two inhibitor factors which suppress lymphocyte proliferation: Characterization and purification of the molecules",
abstract = "The crude supernatant of an CD1+/CD8+ T-cell clone (GI-CO-T-9) established from a long standing culture of an acute T-lymphoblastic leukemia (T-ALL) was shown to inhibit the responsiveness of normal PBMC to PHA. This clone does not produce TNF-α, TNF-β, α-IFN, τ-IFN, IL-1, IL-2 and has no NK-like activity. The crude supernatant has been subjected to a multi-step chromatographic fractioning. Preparative gel permeation HPLC allowed us to recover two peaks of biologic activity in the range of 100-120 kDa and 75-85 kDa (referred to as 'high molecular mass inhibitor factor', HMMIF, and 'low molecular mass inhibitor factor', LMMIF, respectively). Both fractions were then subjected to anion exchange HPLC: HMMIF was recovered in fractions eluting at 0.04 M NaCl while LMMIF eluted at higher ionic strength (0.48 M NaCl). The fractions with biologic activity recovered from anion exchange HPLC have been subjected to hydrophobic interaction HPLC (HIC) for final purification. The highly purified material was characterized by polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate (SDS) revealing single bands of 115 kDa and 80 kDa. The isoelectric points (pI), determined by flat-bed isoelectricfocusing, were 7.4 for HMMIF and 3.5-3.6 for LMMIF. Studies on temperature lability indicated that both proteins are stable for 3-4 hours at room temperature (RT), 24-36 hours at +4°C and 7-10 days at -80°C.",
author = "Montaldo, {P. G.} and M. Lanciotti and E. Castagnola and Parodi, {M. T.} and C. Cirillo and P. Cornaglia-Ferraris and M. Ponzoni",
year = "1988",
language = "English",
volume = "7",
pages = "413--427",
journal = "Lymphokine and Cytokine Research",
issn = "0277-6766",
publisher = "Mary Ann Liebert Inc.",
number = "4",

}

TY - JOUR

T1 - A human acute leukemia-derived T-cell line produces two inhibitor factors which suppress lymphocyte proliferation

T2 - Characterization and purification of the molecules

AU - Montaldo, P. G.

AU - Lanciotti, M.

AU - Castagnola, E.

AU - Parodi, M. T.

AU - Cirillo, C.

AU - Cornaglia-Ferraris, P.

AU - Ponzoni, M.

PY - 1988

Y1 - 1988

N2 - The crude supernatant of an CD1+/CD8+ T-cell clone (GI-CO-T-9) established from a long standing culture of an acute T-lymphoblastic leukemia (T-ALL) was shown to inhibit the responsiveness of normal PBMC to PHA. This clone does not produce TNF-α, TNF-β, α-IFN, τ-IFN, IL-1, IL-2 and has no NK-like activity. The crude supernatant has been subjected to a multi-step chromatographic fractioning. Preparative gel permeation HPLC allowed us to recover two peaks of biologic activity in the range of 100-120 kDa and 75-85 kDa (referred to as 'high molecular mass inhibitor factor', HMMIF, and 'low molecular mass inhibitor factor', LMMIF, respectively). Both fractions were then subjected to anion exchange HPLC: HMMIF was recovered in fractions eluting at 0.04 M NaCl while LMMIF eluted at higher ionic strength (0.48 M NaCl). The fractions with biologic activity recovered from anion exchange HPLC have been subjected to hydrophobic interaction HPLC (HIC) for final purification. The highly purified material was characterized by polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate (SDS) revealing single bands of 115 kDa and 80 kDa. The isoelectric points (pI), determined by flat-bed isoelectricfocusing, were 7.4 for HMMIF and 3.5-3.6 for LMMIF. Studies on temperature lability indicated that both proteins are stable for 3-4 hours at room temperature (RT), 24-36 hours at +4°C and 7-10 days at -80°C.

AB - The crude supernatant of an CD1+/CD8+ T-cell clone (GI-CO-T-9) established from a long standing culture of an acute T-lymphoblastic leukemia (T-ALL) was shown to inhibit the responsiveness of normal PBMC to PHA. This clone does not produce TNF-α, TNF-β, α-IFN, τ-IFN, IL-1, IL-2 and has no NK-like activity. The crude supernatant has been subjected to a multi-step chromatographic fractioning. Preparative gel permeation HPLC allowed us to recover two peaks of biologic activity in the range of 100-120 kDa and 75-85 kDa (referred to as 'high molecular mass inhibitor factor', HMMIF, and 'low molecular mass inhibitor factor', LMMIF, respectively). Both fractions were then subjected to anion exchange HPLC: HMMIF was recovered in fractions eluting at 0.04 M NaCl while LMMIF eluted at higher ionic strength (0.48 M NaCl). The fractions with biologic activity recovered from anion exchange HPLC have been subjected to hydrophobic interaction HPLC (HIC) for final purification. The highly purified material was characterized by polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate (SDS) revealing single bands of 115 kDa and 80 kDa. The isoelectric points (pI), determined by flat-bed isoelectricfocusing, were 7.4 for HMMIF and 3.5-3.6 for LMMIF. Studies on temperature lability indicated that both proteins are stable for 3-4 hours at room temperature (RT), 24-36 hours at +4°C and 7-10 days at -80°C.

UR - http://www.scopus.com/inward/record.url?scp=0024255850&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024255850&partnerID=8YFLogxK

M3 - Article

C2 - 3210814

AN - SCOPUS:0024255850

VL - 7

SP - 413

EP - 427

JO - Lymphokine and Cytokine Research

JF - Lymphokine and Cytokine Research

SN - 0277-6766

IS - 4

ER -