Gutting Bradford W, Updyke Lawrence W, Amacher David E
Groton Laboratories, Pfizer Global Research and Development, Drug Safety Evaluation, Groton, CT 06340, USA.
Toxicol Lett. 2002 May 28;131(3):167-80. doi: 10.1016/s0378-4274(02)00029-2.
Non-steroidal anti-inflammatory drugs (NSAIDs) are frequently associated with immune-mediated hypersensitivity reactions. The NSAID diclofenac is associated with several distinct allergic and autoimmune-like reactions including anaphylaxis, idiosyncratic hepatotoxicity and autoimmune hemolytic anemia. The aim of this study was to examine the immunostimulating potential of diclofenac in the direct popliteal lymph node assay (PLNA) and reporter antigen PLNA. In BALB/c mice, diclofenac caused dose-dependent increases in PLN weight and PLN cellularity in the direct PLNA; 0.25 mg was non-immunostimulating whereas 0.50-1.00 mg caused a significant PLN reaction. In the direct PLNA, diclofenac also increased the percent of T cells in the PLN with activated phenotypes (CD44(high)CD62L(low) and CD44(high)CD62L(high)). Finally, the magnitude of the diclofenac-induced direct PLN reaction was significantly reduced when the assay was conducted in T-cell-deficient mice. When co-injected with the reporter antigen TNP-Ficoll (trinitrophenyl Ficoll), 0.50 mg diclofenac caused significant increases in PLN weight, PLN cellularity, and induced IgM and IgG(1) anti-TNP antibody forming cells (AFCs) in the PLN. In a final set of studies, a TNP-OVA PLNA was conducted using diclofenac, phenobarbital (negative control) and streptozotocin (positive control). As expected, phenobarbital (1.00 mg) failed to cause an increase in PLN cellularity or induce AFCs in the PLN. Streptozotocin (1.00 mg) caused significant increases in PLN cellularity, IgM AFCs, and selectively induced IgG(2a) and IgG(2b) AFCs against TNP-OVA. Likewise, diclofenac caused dose-dependent increases (0.25-1.00 mg) in PLN cellularity and IgM AFCs. However, in contrast to streptozotocin, diclofenac caused a selective dose-dependent increase in both IgG(1) and IgE AFCs. Finally, an increase in the intracellular level of IL-4, but not INFgamma, was detected in CD4(+) PLN cells following the injection of diclofenac mixed with TNP-OVA. Collectively, these data suggest that diclofenac: (i) induces a T-cell-dependent direct PLN reaction that; (ii) provides non-cognate help for IgG AFC production when co-injected with TNP-Ficoll, possibly through the formation of neo-antigens; and (iii) possesses intrinsic adjuvant activity that selectively induces IL-4 mediated production of IgG(1) and IgE against co-injected TNP-OVA.
非甾体抗炎药（NSAIDs）常与免疫介导的超敏反应相关。NSAID双氯芬酸与多种不同的过敏和自身免疫样反应有关，包括过敏反应、特异质性肝毒性和自身免疫性溶血性贫血。本研究的目的是在直接腘窝淋巴结试验（PLNA）和报告抗原PLNA中检测双氯芬酸的免疫刺激潜力。在BALB/c小鼠中，双氯芬酸在直接PLNA中导致腘窝淋巴结重量和细胞数量呈剂量依赖性增加；0.25mg无免疫刺激作用，而0.50 - 1.00mg引起显著的腘窝淋巴结反应。在直接PLNA中，双氯芬酸还增加了具有活化表型（CD44(高)CD62L(低)和CD44(高)CD62L(高)）的T细胞在腘窝淋巴结中的百分比。最后，当在T细胞缺陷小鼠中进行试验时，双氯芬酸诱导的直接腘窝淋巴结反应的强度显著降低。当与报告抗原三硝基苯 - 聚蔗糖（TNP - Ficoll）共同注射时，0.50mg双氯芬酸导致腘窝淋巴结重量、细胞数量显著增加，并诱导腘窝淋巴结中IgM和IgG(1)抗TNP抗体形成细胞（AFCs）。在最后一组研究中，使用双氯芬酸、苯巴比妥（阴性对照）和链脲佐菌素（阳性对照）进行了TNP - OVA PLNA试验。正如预期的那样，苯巴比妥（1.00mg）未能导致腘窝淋巴结细胞数量增加或诱导腘窝淋巴结中的AFCs。链脲佐菌素（1.00mg）导致腘窝淋巴结细胞数量、IgM AFCs显著增加，并选择性诱导针对TNP - OVA的IgG(2a)和IgG(2b) AFCs。同样，双氯芬酸导致腘窝淋巴结细胞数量和IgM AFCs呈剂量依赖性增加（0.25 - 1.00mg）。然而，与链脲佐菌素不同的是，双氯芬酸导致IgG(1)和IgE AFCs均呈选择性剂量依赖性增加。最后，在注射与TNP - OVA混合的双氯芬酸后，在CD4(+)腘窝淋巴结细胞中检测到IL - 4细胞内水平升高，但INFγ未升高。总体而言，这些数据表明双氯芬酸：（i）诱导T细胞依赖性直接腘窝淋巴结反应；（ii）当与TNP - Ficoll共同注射时，可能通过新抗原的形成，为IgG AFC产生提供非同源辅助；（iii）具有内在佐剂活性，可选择性诱导IL - 4介导的针对共同注射的TNP - OVA的IgG(1)和IgE产生。
