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M9610807.TXT
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1996-01-30
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Document 0807
DOCN M9610807
TI Characterization of latent transforming growth factor-beta from human
seminal plasma.
DT 9601
AU Nocera M; Chu TM; Department of Diagnostic Immunology Research and
Biochemistry,; Roswell Park Cancer Institute, New York State Department
of; Health, Buffalo 14263, USA.
SO Am J Reprod Immunol. 1995 Apr;33(4):282-91. Unique Identifier : AIDSLINE
MED/96037778
AB PROBLEM: Human seminal plasma is known to exhibit immunosuppressive
activity. Transforming growth factor beta (TGF-beta) has been identified
as an immunosuppressive factor in human seminal plasma. Biologically
active TGF-beta represents a family of 25-kDa homodimeric proteins
linked with disulfide bonds. TGF-beta associates with high molecular
weight proteins noncovalently to form a type of latency that is
biologically inactive. Quantitative distribution of active form of
TGF-beta versus inactive latent form of of TGF-beta, and mechanism of
the TGF-beta activation in human seminal plasma remain to be elucidated.
PURPOSE: To characterize seminal plasma latent form of TGF-beta,
including its concentration, and the mechanism underlying the activation
of TGF-beta. METHOD: Gel filtrations on ACA-34 and Biogel P-60 were used
to fractionate seminal plasma. TGF-beta was measured by enzyme
immunoassay using antibodies specific for TGF-beta 1 and TGF-beta 2,
respectively. Radioreceptor assay with recombinant human [125I]-TGF-beta
1 was applied to qualitatively identify TGF-beta 1. Kinetic experiments
with various pH, temperature and time, along with protease inhibitors,
were performed to delineate the activation mechanism of latent TGF-beta
1. RESULTS: Human seminal plasma contained both TGF-beta 1 and TGF-beta
2, predominantly in latent form. The total concentration of TGF-beta 1
averaged 238 ng/ml versus an average of 18 ng/ml for TGF-beta 2. The in
vitro activation or release of TGF-beta 1 from latent TGF-beta 1 was
achieved only at acidic pH of < 4.0, and was time and temperature
dependent. At pH 3.7 and 37 degrees C, a significant activation of
latent TGF-beta 1 was achieved after an incubation of only 15 min,
reached the maximum at 120 min, and the activated TGF-beta 1 remained
relatively stable for at least 24 h. The activation was not inhibitable
by a series of protease inhibitors examined, alone or in combination
(e.g., phenyl-methylsulfonyl fluoride, E-64, pepstatin, leupeptin,
ethylenediamine tetraacetic acid). Competitive radioreceptor assay
established the functional identity of TGF-beta 1 in human seminal
plasma with recombinant human TGF-beta 1. CONCLUSION: Human seminal
plasma TGF-beta is biologically activated from high molecular weight
latent TGF-beta by acid pH. The acidic environment of female lower
genital tract could represent an in vivo physiological condition for
activation of seminal plasma TGF-beta that may immunologically protect
the integrity of sperm.
DE Animal Chromatography Female Genitalia, Female/METABOLISM Human
Hydrogen-Ion Concentration HIV Infections/TRANSMISSION Immunoenzyme
Techniques Male Mink Reference Standards
Semen/*CHEMISTRY/IMMUNOLOGY/PHYSIOLOGY Transforming Growth Factor
beta/*ANALYSIS/IMMUNOLOGY/ *PHARMACOKINETICS JOURNAL ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).