Inhibition of the Secondary Antibody Response in Vitro by Salicylate and Gentisate

Abstract

Salicylate inhibition of the secondary antibody response initiated in vitro on day 0 has been studied in cultures of rabbit lymph node fragments. Levels of 1.25 to 1.5 mM (0.20 to 0.24 mg/ml) sodium salicylate present in serum-free medium throughout an 18- or 21-day culture period completely inhibit the secondary response. This inhibition is largely accomplished by the drug's action during the first 9 days, which corresponds to the inductive phase for this culture system. Relatively little inhibition is produced by adding the drug only after day 9, although over 90% of the antibody produced during a 21-day experiment is synthesized after day 9. Studies with media of different pH's show that this inhibition is more correctly a function of the nonionized salicylic acid concentration in the medium than of the total salicylate concentration. Arguments are presented against the possibility that salicylate at the levels used here inhibits antibody synthesis by uncoupling oxidative phosphorylation. Acetylsalicylic acid (aspirin) produces the same degree of inhibition in vitro as do equimolar concentrations of sodium salicylate. Gentisate (5-hydroxysalicylate) is 15-fold more effective in producing 50% inhibition than salicylate; its temporal pattern of inhibition is similar to that of salicylate.

Effect of antipyretic analgesics on immune responses to vaccination

“…In a rabbit spleen culture system, antibody suppression by salicylates was noted mainly during the inductive phase of the culture system (first 9 days), with very little effect afterwards; although most of the antibody production was noticed after the inductive period. 23 In similar studies using human peripheral blood mononuclear cells (PBMCs), indomethacin maximally inhibited immunoglobulin secretion when the drug was added early on or in the first 24 hours of culture. [46][47][48] Ibuprofen added during the first days of PBMC culture (days 1, 2 and/or 3) produced greater IgM suppression versus adding the drug at later time points (day 5 and/or day 6).…”

“…Salicylate suppressed immunoglobulin production in a dose-dependent manner. 23 Similarly, salicylate inhibited thymidine incorporation and decreased the number of antibody forming cells found in spleen cells cultured from chickens immunized with sheep red blood cells (SRBCs). 24 However, other investigators reported differing results.…”

Regulation of the Secondary Antibody Response in Vitro

“…Our modifications of this technique, used in current experiments and described in previous papers (3)(4)(5), include the following: (a) the development of a serum-free, chemically defined medium; (b) the replacement of the plasma clot by a glass wool pad overlying the fragments in a Leighton tube, thus fixing them in position; (c) the conversion to stationary cultures; (aT) the control of pH by gassing individual tubes with 5% COS in air; (e) the utilization of other lymph nodes, such as the axillary, auricular, and cervical nodes; and (.f) the use of other antigens, including tetanus toxoid and ovalbumin. The several modifications not previously described but used in experiments of this paper are discussed below in some detail.…”