Binding of modified alpha-1-antichymotrypsin to mitogen-stimulated human lymphocyte membrane: a model for immune suppression.
Alpha-1-antichymotrypsin (ACT), an acute phase reactant protein elevated during acute inflammation, and its derivatives (asialo ACT and acid-exposed asialo ACT) were investigated their effect on lymphocyte proliferative responses, and evidence for binding to lymphocyte membranes as well as the characteristics of this binding were investigated. Acid-exposed asialo ACT significantly reduced 3H-thymidine incorporation into human peripheral lymphocytes stimulated by phytohemagglutinin (PHA) though native ACT could not inhibit the mitogen-induced lymphoproliferation and asialo ACT moderately inhibited it. In order to determine the interaction of ACT and its derivatives to lymphocyte membranes, the binding of 125I-labelled ACT and its derivatives to membranes of intact lymphocyte and extracted lymphocyte membranes was examined. The binding of 125I-labelled native ACT and asialo ACT to resting and PHA-stimulated lymphocyte membrane was low. And the binding of 125I-labelled acid-exposed asialo ACT to resting lymphocyte membrane was also low. However, when lymphocytes were stimulated by mitogens the binding of 125I-labelled acid-exposed asialo ACT increased significantly. The binding of 125I-labelled acid-exposed asialo ACT to the membrane extracted from PHA-stimulated lymphocytes was time-dependent and saturation was reached at 120 min at 37 degrees C. One mg of membrane could bind a maximum of approximately 83 pmol of acid-exposed asialo ACT with dissociation constant of 0.73 microM. Other unlabelled serum glycoproteins such alpha-1-antitrypsin, alpha-1-acid glycoprotein, transferrin, and proteinase inhibitors including chymostatin, leupeptin and soy bean trypsin inhibitor did not compete with 125I-labelled acid-exposed asialo ACT for binding sites in simultaneous competition assays.
Matsumoto
M
M
Department of Biochemistry, School of Medicine, Tokai University, Kanagawa, Japan.
Yamamura
M
M
Takada
S
S
Nakamura
K
K
Katsunuma
T
T
D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007108:Immune Tolerance / Q000502:physiology*; D066298:In Vitro Techniques; D007700:Kinetics; D008213:Lymphocyte Activation / Q000502:physiology; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism*; D008934:Mitogens / Q000494:pharmacology; D000514:alpha 1-Antichymotrypsin / Q000276:immunology / Q000378:metabolism*
D008934:Mitogens; D000514:alpha 1-Antichymotrypsin