T cell cytokine pattern in the joints of patients with lyme arthritis and its regulation by cytokines and anticytokines

Abstract

Objective. To investigate whether type 1 helper (Thl) or Th2 cytokines are found in the joints of patients with Lyme arthritis, and whether the cytokine pattern can be modulated by cytokines or anticytokines.Methods. The cytokine pattern in the joints of 10 patients with Lyme arthritis was investigated. Expression of interferon-y (IFN y), tumor necrosis factor a (TNFa), interleukin-4 (IL-4), and IL-10 was measured by enzyme-linked immunosorbent assay (ELISA), after stimulation of synovial fluid mononuclear cells (SFMC) with Borreliu burgdorferi (Bb) in the supernatant. Expression of cytokine messenger RNA and protein in synovial membrane (SM) and nonstimulated SFMC was studied using semiquantitative reverse transcriptasepolymerase chain reaction and immunohistologic techniques. The effects of recombinant cytokines or neutralizing anticytokine antibodies on cytokine production in Bb-stimulated SFMC were investigated by ELISA.Results. SFMC produced high amounts of IFNy and TNFa, but little or no IL-4, upon stimulation with Bb antigen, indicating a Thl-type cytokine pattern. In SM, IFNy was detectable in all patients, while the other cytokines were less frequently found. Serial sections of SM revealed that all cytokines were located in the same area. The Thl response, especially the production of

Crucial role of interleukin‐10/interleukin‐12 balance in the regulation of the type 2 T helper cytokine response in reactive arthritis

“…A semiquantitative PCR for different IFNy genes was performed as described previously (25,26). In a competitive PCR, the amount of cDNA was quantified using specific primers for p-actin and using a control fragment.…”

“…Five microliters of the PCR product was electrophoresed through a 1.5% agarose gel and visualized by ethidium bromide staining. Primer sequences, annealing temperature, and the length of the amplified fragments have been previously described (26).…”

Multispecific CD4+ T Cell Response to a Single 12-mer Epitope of the Immunodominant Heat-Shock Protein 60 ofYersinia enterocoliticainYersinia-Triggered Reactive Arthritis: Overlap with the B27-Restricted CD8 Epitope, Functional Properties, and Epitope Presentation by Multiple DR Alleles

“…Culture supernatants were harvested after 48 h of T cell stimulation for determination of Ag-induced cytokine secretion, and the concentration of cytokines in the supernatant was measured as described previously (5,22). Briefly, levels of IFN-␥, TNF-␣, IL-10, and IL-4 were analyzed by sandwich ELISA using purified anti-cytokine mAb (capture) and biotinylated anti-cytokine mAbs (detecting) (PharMingen, San Diego, CA), with a detection level of 6 pg/ml for IL-4 and 20 pg/ml for the other three cytokines.…”

Interleukin-10 (IL-10) Inhibits Borrelia burgdorferi-Induced IL-17 Production and Attenuates IL-17-Mediated Lyme Arthritis

“…Until now, much of the investigation pertaining to the inflammatory targets of IL-10 focused on the Th1/IFN-␥ response, which has long been known as a major inflammatory pathway of later-stage Lyme arthritis (5,6). For example, modulation of IL-10 in vitro reciprocally influenced IFN-␥ production by stimulated synovial cells of Lyme arthritis patients (29). In addition, differences in IL-10-mediated regulation of IFN-␥ accounted for the differences in arthritic severity exhibited by C3H and C57BL/6 mice (3,28).…”

“…In contrast, the effects of IL-10 on IL-17 production were more pronounced after 1 week of infection. Borrelial infection in the presence or absence of IL-10 may be associated with an inverse response of various inflammatory cytokines, including not only tumor necrosis factor alpha and IFN-␥ (3,4,(28)(29)(30)(46)(47)(48)(49)(50)(51) but also IL-17. If a certain threshold of immunosuppression by IL-10 is not achieved, a significant, multifaceted inflammatory response may ensue.…”