Clin. exp. Immunol (1991) 83, 516-517

ADONIS 000991049100094J

Letter to the Editors Peripheral blood monocytes from patients with reactive arthritis show normal production of tumour necrosis factor-alpha

We studied 15 patients (13 HLA-B27+ positive and two HLA-B27-), mean age 33 years (range 16-45): eight with chlamydia urethritis-triggered ReA, two with yersinia enteritistriggered ReA, one with campylobacter-triggered ReA, and four with non-specific enteritis-triggered ReA at the acute phase of the disease. Control subjects, mean age 37 years (range 20-48) were age- and sex-matched, HLA-B27- laboratory and hospital personnel. A patient and a control were studied concurrently, and 13 out of 15 pairs were restudied 6 months later. At the acute phase/at 6 months, mean maximum erythrocyte sedimentation rate (range) was 80 (22-115)/7 (2-12) mm/h, mean maximum Creactive protein level 85 (13-206)/6 (2-14) mg/i, and mean maximum leucocyte count 9-7 (5-9-14-7)/7-1 (3-6-8A4) x 10-9/l. Buffy coat was separated from heparinized blood by dextran

Sirs, Reactive arthritis (ReA), a sterile joint inflammation, can be triggered by enteric infections caused by salmonella, shigella, campylobacter, or yersinia, and by urogenital infections caused by chlamydia. About 80% of the patients are HLA-B27 positive Chlamydia (Keat et al., 1987), yersinia (Granfors et al., 1989) and salmonella (Granfors et al., 1990a) antigens have been detected in the joints of the patients with ReA triggered by the corresponding microbes. In a recent study of patients with Yersinia enterocolitica 0.3 infection, peripheral blood cells of almost every patient studied contained yersinia antigens (Granfors et al., 1990b), suggesting that aberrant host response to the 12 r ( a)

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Fig. 1. Production of TNF-a by monocytes of patients (0) and controls (0) at the acute phase (a) of reactive arthritis (n = 15) and 6 months later (b) (n = 13). LPS, lipopolysaccharide. Horizontal lines indicate arithmetic means.

microbial components, rather than occurrence of antigenaemia, would determine the development of reactive arthritis. The gene for tumour necrosis factor-alpha (TNF-a) is a part of the human histocompatibility complex close to the HLA-B locus (Spies et al., 1986) and might thereby play a role in the pathogenesis of HLA-B27-associated disorders. This possibility is supported by our previous findings that mononuclear cells of healthy subjects who are HLA-B27+ and of patients with previous yersinia ReA generate more TNF-cx in response to Escherichia coli lipopolysaccharide (LPS) than do control cells (Repo et al., 1988). The difference might have derived from increased generation of TNF-x by monocytes separated by means of plastic adherence, or from a higher cell count due to increased adherence of the cells. This prompted us to study production of TNF-at by a fixed number of monocytes from patients with ReA at the early phase of the disease and after the patients had recovered. Correspondence: H. Reppo, Department of Bacteriology and Immunology, University of Helsinki, Haartmaninkatu 3, SF-00290, Helsinki, Finland.

sedimentation, mononuclear cells from buffy coat by FicollIsopaque density grandient centrifugation, and monocytes (8095% pure) from mononuclear cells by Percoll density gradient centrifugation. To remove platelets, monocyte suspension was layered on platelet-poor pooled human plasma, centrifuged (50 g for 10 min), and washed. Cell viability by eosin exclusion was >90%. Monocytes (2-5 x 105) in 500 pl of RPMI 1640 (GIBCO, Paisley, UK) containing 10% of heat-inactivated human AB serum were exposed to E. coli 026.B6 LPS (Difco, Detroit, MI.) 10-0 pg/ml, and cultured for 20 h on a 24-well plate (Costar, no. 3424, Cambridge, MA) at 37°C in 5% C02 in air. The levels of TNF-a in the culture supernatants were determined by using radioimmunoassay, as described in detail (Repo et al., 1988). This assay detects both biologically active and inactive TNF-a (Jaittela, Kuusela & Saksela, 1988). LPS-induced TNF-at production from monocytes in a dosedependent manner (Fig. 1), and the peak responses were always obtained with LPS 10 yg/ml. The patients' responses at the acute phase of the disease were similar to those after recovery, and 516

Letter to the Editors they were consistantly similar to those of the controls. The findings disagree with our previous results indicating that both the patients with previous yersinia-reactive arthritis and HLAB27+ subjects without a history of ReA show enhanced TNF-i production (Repo et al., 1988). The discrepancy may be because (i) differences between the separation methods of adherent cells, such as differences in enrichment of subpopulations of monocytes (Szabo et al., 1990) or in adhesion properties of the cells; (ii) a too low number of subjects studied; or (iii) other factors. The data give credence to the view that there are no major aberrations in LPS-induced TNF-a production of monocytes from patients with ReA. Keywords reactive arthritis tumour necrosis factor-alpha

H. Repo*§, A. Lauhio*, M. Jaattelat, P. Saikkut & M. Leirisalo-Repo*§ * Departments of * Bacteriology and Immunology, t Pathology, : Virology, and § Medicine II, University of Helsinki, Helsinki, Finland. REFERENCES GRANFORS, K., JALKANEN, S., VON ESSEN. R., LAHESMAA-RANTALA, R., ISOMAKI, O., PEKKOLA-HEINO, K., MERILAHTI-PALO, R., SAARIO, R., ISOMAKI, H. & TOIVANEN, A. (1989) Yersinia antigens in synovialfluid cells from patients with reactive arthritis. N. Engl J. Med. 320, 216.

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GRANFORS, K., JALKANEN, S., LINDBERG, A.A., MAKI-IKOLA, O., VON ESSEN, R., LAHESMAA-RANTALA, R., ISOMKKI, H., SAARIO, R., ARNOLD, W.J. & TOIVANEN, A. (1990a) Salmonella lipopolysaccharide in synovial cells from patients with reactive arthritis. Lancet, 335, 685. GRANFORS, K., JALKANEN, S., LAHESMAA-RANTALA, R., SAARIO, R., MTrTONEN, T. & TOIVANEN, A. (1990c) Bacterial antigens in peripheral blood cells in yersinia-triggered reactive arthritis. Scand. J. Rheumatol. Suppl. 85, 45 (Abstract). JAATTELA, M., KUUSELA, P. & SAKSELA, E. (1988) Demonstration of tumor necrosis factor in human amniotic fluids and supernatants of placental and decidual tissues. Lab. Invest. 58, 48. KEAT, A., THOMAS, B., DIxEY, J., OSBORN, M., SONNEX, C. & TAYLORROBINSON, D. (1987) Chlamydia trachomatis and reactive arthritis: the missing link. Lancet 1, 72. REPO, H., JAATTELA, M., LEIRISALO-REPO, M. & HURME, M. (1988)

Production of tumour necrosis factor and interleukin- 1 by monocytes of patients with previous yersinia arthritis. Clin. exp. Immunol. 72, 410. SPIES, T., MORTON, C.C., NEDOSPASOV, S.A., FIERS, W., Pious, D. & STROMINGER, J.L. (1986) Genes for the tumor necrosis factors a and /3 are linked to the human major histocompatibility complex. Proc. natl Acad. Sci. USA, 83, 8699. SZABO, G., MILLER-GRAZIANO, C.L., Wu, J.-Y., TAKAYAMA, T. & KODYS, K. (1990) Differential tumor necrosis factor production by human monocyte subsets. J. Leukocyte Biol. 47, 206.

Peripheral blood monocytes from patients with reactive arthritis show normal production of tumour necrosis factor-alpha.

Clin. exp. Immunol (1991) 83, 516-517 ADONIS 000991049100094J Letter to the Editors Peripheral blood monocytes from patients with reactive arthritis...
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