Inhibition of cytotoxicity by sulfasalazine. II. Sulfasalazine and sulfapyridine inhibit different stages of the NK and NKCF lytic processes

doi:10.1016/0162-3109(86)90031-7

Immunopharmacology

Volume 11, Issue 2, April 1986, Pages 111-118

Immunopharmacology

https://doi.org/10.1016/0162-3109(86)90031-7 Get rights and content

Abstract

Sulfasalazine and sulfapyridine but not 5-aminosalicylate inhibit spontaneous cytotoxicity mediated by human natural killer (NK) cells. The aim of this study was to determine which stage(s) of the NK cytotoxic reaction is inhibited by these compounds. Effector/target cell binding studies performed in parallel with cytoxicity assays using purified large granular lymphocytes indicated that inhibition is a post-binding event. The kinetic profile of inhibition in a calcium pulse assay showed that inhibition continues long after the effector cell triggering stage and that although sulfasalazine may have some inhibitory effect on the calcium-dependent events of the programming phase, sulfapyridine continues to inhibit during the calcium-independent or lethal hit phase of the cytotoxic sequence. The NK soluble cytotoxic factor (NKCF) assay was used as a measure of the lethal hit since the time course of this assay permits study of the various substages of this terminal event in the lytic sequence. Sulfasalazine and sulfapyridine but not 5-aminosalicylate inhibited NKCF-mediated target cell lysis. Different substages of the NKCF-induced lytic reaction were affected by these agents. Sulfasalazine appears to inhibit binding of NKCF to the target cell whereas sulfapyridine predominantly inhibits early post-binding events.

References (16)

S.R. Targan et al.
NK-Target cell interactions in binding, triggering, programming, and lethal hit stages of NK cytotoxicity
T. Timonen et al.
Isolation of human natural killer cells by density gradient centrifugation
J Immunol Methods
(1980)
A. Boyum
Isolation of leukocytes from human blood. A two-phase system for removal of red cells with methylcellulose as erythrocyte aggregating agent
Scand J Clin Lab Invest
(1968)
J.A. Brieva et al.
NK and T cell subsets regulate antibody production by human in vivo antigen-induced lymphoblastoid B cells
J Immunol
(1984)
R.L. Deem et al.
Evidence of a dynamic role of the target cell membrane during the early stages of the natural kill cell lethal hit
J Immunol
(1984)
R.L. Deem et al.
Sequential substages of natural killer cell-derived cytolytic factor (NKCF)-mediated cytolysis as defined by glutaraldehyde modulation of the target cell
J Immunol
(1984)
E. Farram et al.
Identification of human natural killer soluble cytotoxic factor(s) (NKCF) derived from NK-enriched lymphocyte populations: specificity of generation and killing
J Immunol
(1983)
P. Goldstein et al.
The lethal hit stage of mouse T and non-T cell-mediated cytolysis: Differences in cation requirements and characterization of an analytical ‘cation pulse’ method
Eur J Immunol
(1976)

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Immunopharmacology

Abstract

References (16)

NK-Target cell interactions in binding, triggering, programming, and lethal hit stages of NK cytotoxicity

Isolation of human natural killer cells by density gradient centrifugation

J Immunol Methods

Isolation of leukocytes from human blood. A two-phase system for removal of red cells with methylcellulose as erythrocyte aggregating agent

Scand J Clin Lab Invest

NK and T cell subsets regulate antibody production by human in vivo antigen-induced lymphoblastoid B cells

J Immunol

Evidence of a dynamic role of the target cell membrane during the early stages of the natural kill cell lethal hit

J Immunol

Sequential substages of natural killer cell-derived cytolytic factor (NKCF)-mediated cytolysis as defined by glutaraldehyde modulation of the target cell

J Immunol

Identification of human natural killer soluble cytotoxic factor(s) (NKCF) derived from NK-enriched lymphocyte populations: specificity of generation and killing

J Immunol

The lethal hit stage of mouse T and non-T cell-mediated cytolysis: Differences in cation requirements and characterization of an analytical ‘cation pulse’ method

Eur J Immunol

Cited by (19)

Miscellaneous treatments, I: Sulfasalazine and pentoxifylline: Unapproved uses, dosages, or indications

Progress in understanding the mechanisms of action of 5-aminosalicylic acid

Sulfasalazine inhibits the binding of TNFα to its receptor

Inhibition of Proliferative Responses and Interleukin 2 Productions by Salazosulfapyridine and Its Metabolites

Inhibition of antibody secretion by 5-aminosalicylic acid

Alterations of the Immune System in Ulcerative Colitis and Crohn's Disease