Partial purification and characterization of a lymphocyte-inhibitory factor(s) in ascitic fluids from ovarian cancer patients

A. D. Hess, S. A. Gall, J. R. Dawson

Research output: Contribution to journalArticle

Abstract

Attempts were undertaken to purify and characterize the lymphocyte-inhibitory activity found in the ascitic effusions from ovarian cancer patients. The active moiety responsible for the inhibition of in vitro lymphocyte function was partially purified from several ascitic fluids by means of concanavalin A-Sepharose 4B (negative) affinity and DE52 ion-exchange chromatography, with final yields of 68 to 74% and 3- to 8-fold increases in specific activity, Analytical polyacrylamide gel electrophoresis showed that the active fraction contained mostly albumins and α-globulins. Further purification by semi-preparative electrophoresis in polyacrylamide gels revealed that the majority of the lymphocyte-inhibitory activity migrated with the albumins, although minor levels of activity were detected in the material which eluted from the α-globulin region. The lymphocyte-inhibitory activity was found to be separable from albumin by means of a rabbit anti-human serum albumin immunoadsorbent column. The active factor which lost activity upon storage at -20° was found to have a molecular size of 40,000 to 80,000 daltons, as estimated by Sephadex G-200 filtration. Attempts to separate an active, small-molecular-size component from fractions containing the inhibitory activity by acid ultrafiltration were largely unsuccessful. Immunodiffusion analysis of the purified fractions following removal of albumin revealed the presence of contaminating amounts of α1-acid glycoprotein and α1-antitrypsin. However, levels of these two proteins did not correlate with the presence of lymphocyte-inhibitory activity.

Original languageEnglish (US)
Pages (from-to)1842-1851
Number of pages10
JournalCancer Research
Volume40
Issue number6
StatePublished - 1980

Fingerprint

Ascitic Fluid
Ovarian Neoplasms
Lymphocytes
Albumins
Globulins
Polyacrylamide Gel Electrophoresis
Immunosorbents
Acids
Immunodiffusion
Ion Exchange Chromatography
Ultrafiltration
Serum Albumin
Sepharose
Glycoproteins
Rabbits
Proteins

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Partial purification and characterization of a lymphocyte-inhibitory factor(s) in ascitic fluids from ovarian cancer patients. / Hess, A. D.; Gall, S. A.; Dawson, J. R.

In: Cancer Research, Vol. 40, No. 6, 1980, p. 1842-1851.

Research output: Contribution to journalArticle

@article{2f55fa37b06f41ca843bb412ab5332ac,
title = "Partial purification and characterization of a lymphocyte-inhibitory factor(s) in ascitic fluids from ovarian cancer patients",
abstract = "Attempts were undertaken to purify and characterize the lymphocyte-inhibitory activity found in the ascitic effusions from ovarian cancer patients. The active moiety responsible for the inhibition of in vitro lymphocyte function was partially purified from several ascitic fluids by means of concanavalin A-Sepharose 4B (negative) affinity and DE52 ion-exchange chromatography, with final yields of 68 to 74{\%} and 3- to 8-fold increases in specific activity, Analytical polyacrylamide gel electrophoresis showed that the active fraction contained mostly albumins and α-globulins. Further purification by semi-preparative electrophoresis in polyacrylamide gels revealed that the majority of the lymphocyte-inhibitory activity migrated with the albumins, although minor levels of activity were detected in the material which eluted from the α-globulin region. The lymphocyte-inhibitory activity was found to be separable from albumin by means of a rabbit anti-human serum albumin immunoadsorbent column. The active factor which lost activity upon storage at -20° was found to have a molecular size of 40,000 to 80,000 daltons, as estimated by Sephadex G-200 filtration. Attempts to separate an active, small-molecular-size component from fractions containing the inhibitory activity by acid ultrafiltration were largely unsuccessful. Immunodiffusion analysis of the purified fractions following removal of albumin revealed the presence of contaminating amounts of α1-acid glycoprotein and α1-antitrypsin. However, levels of these two proteins did not correlate with the presence of lymphocyte-inhibitory activity.",
author = "Hess, {A. D.} and Gall, {S. A.} and Dawson, {J. R.}",
year = "1980",
language = "English (US)",
volume = "40",
pages = "1842--1851",
journal = "Journal of Cancer Research",
issn = "0099-7013",
publisher = "American Association for Cancer Research Inc.",
number = "6",

}

TY - JOUR

T1 - Partial purification and characterization of a lymphocyte-inhibitory factor(s) in ascitic fluids from ovarian cancer patients

AU - Hess, A. D.

AU - Gall, S. A.

AU - Dawson, J. R.

PY - 1980

Y1 - 1980

N2 - Attempts were undertaken to purify and characterize the lymphocyte-inhibitory activity found in the ascitic effusions from ovarian cancer patients. The active moiety responsible for the inhibition of in vitro lymphocyte function was partially purified from several ascitic fluids by means of concanavalin A-Sepharose 4B (negative) affinity and DE52 ion-exchange chromatography, with final yields of 68 to 74% and 3- to 8-fold increases in specific activity, Analytical polyacrylamide gel electrophoresis showed that the active fraction contained mostly albumins and α-globulins. Further purification by semi-preparative electrophoresis in polyacrylamide gels revealed that the majority of the lymphocyte-inhibitory activity migrated with the albumins, although minor levels of activity were detected in the material which eluted from the α-globulin region. The lymphocyte-inhibitory activity was found to be separable from albumin by means of a rabbit anti-human serum albumin immunoadsorbent column. The active factor which lost activity upon storage at -20° was found to have a molecular size of 40,000 to 80,000 daltons, as estimated by Sephadex G-200 filtration. Attempts to separate an active, small-molecular-size component from fractions containing the inhibitory activity by acid ultrafiltration were largely unsuccessful. Immunodiffusion analysis of the purified fractions following removal of albumin revealed the presence of contaminating amounts of α1-acid glycoprotein and α1-antitrypsin. However, levels of these two proteins did not correlate with the presence of lymphocyte-inhibitory activity.

AB - Attempts were undertaken to purify and characterize the lymphocyte-inhibitory activity found in the ascitic effusions from ovarian cancer patients. The active moiety responsible for the inhibition of in vitro lymphocyte function was partially purified from several ascitic fluids by means of concanavalin A-Sepharose 4B (negative) affinity and DE52 ion-exchange chromatography, with final yields of 68 to 74% and 3- to 8-fold increases in specific activity, Analytical polyacrylamide gel electrophoresis showed that the active fraction contained mostly albumins and α-globulins. Further purification by semi-preparative electrophoresis in polyacrylamide gels revealed that the majority of the lymphocyte-inhibitory activity migrated with the albumins, although minor levels of activity were detected in the material which eluted from the α-globulin region. The lymphocyte-inhibitory activity was found to be separable from albumin by means of a rabbit anti-human serum albumin immunoadsorbent column. The active factor which lost activity upon storage at -20° was found to have a molecular size of 40,000 to 80,000 daltons, as estimated by Sephadex G-200 filtration. Attempts to separate an active, small-molecular-size component from fractions containing the inhibitory activity by acid ultrafiltration were largely unsuccessful. Immunodiffusion analysis of the purified fractions following removal of albumin revealed the presence of contaminating amounts of α1-acid glycoprotein and α1-antitrypsin. However, levels of these two proteins did not correlate with the presence of lymphocyte-inhibitory activity.

UR - http://www.scopus.com/inward/record.url?scp=0018860783&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0018860783&partnerID=8YFLogxK

M3 - Article

C2 - 6966184

AN - SCOPUS:0018860783

VL - 40

SP - 1842

EP - 1851

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0099-7013

IS - 6

ER -