Clin. exp. Immunol. (1990) 80, 444-447
Modulation of leucocyte locomotion by interleukin-1 C. C. ZIELINSKI, B. PESAU, W. KALINOWSKI & C. MULLER* Second Department of Medicine, and *Second Department of Gastroenterology and Hepatology, University Hospital, and Ludwig Boltzmann Institut fur prdnatale und experimentelle Genomanalytik, Vienna, Austria (Acceptedfor publication 4 December 1989)
SUMMARY The influence of interleukin-1 (IL-1) upon leucocyte locomotion in vitro was studied, using either casein or zymosan-activated serum (ZAS) as chemotaxigens. A pre-incubation of polymorphonuclear cells (PMNC) with ultrapure (purity 99%) human IL-I (1 U/ml) for 2 and 5 h followed by a washing step resulted in a significant decrease in leucocyte locomotion against casein (P < 0 0005 and P0-1). Moreover, the direct addition of this IL-I preparation to leucocyte locomotion assays without pre-incubation produced a similar and significant inhibition of leucocyte locomotion directed against casein (P < 0 05). This inhibitory effect could not be augmented further by higher concentrations of ultrapure IL-i. In order to exclude the effect of possibly contaminating cytokines in the ultrapure IL- 1 preparation used, additional assays with recombinant human IL-I (rIL-I) alpha and rIL- 1 beta (I U/ml) were performed to investigate their influence upon leucocyte locomotion. It was found that both rIL-la and rIL-lB inhibited leucocyte locomotion directed against casein significantly (P95% PMNC. Leucocyte locomotion assays Leucocyte locomotion was performed using modified Boyden chambers (Boyden, 1962; Frei, Baisero & Ochsner, 1974), as described previously (Zielinski et al., 1979). Briefly, 0-5 ml of the cell suspension containing 2-8 x106 PMNC/ml RPMI 1640 supplemented with antibiotics were pipetted into the upper
Correspondence: Christoph C. Zielinski, MD, Second Department of Medicine, University Hospital, 13 Garnisongasse, A-1090 Vienna, Austria.
444
Leucocyte locomotion inhibition by IL-4
445
Table 1. Inhibition of leucocyte locomotion by ultrapure IL-I (I U/ml) in vitro (chemotaxigen 0.5% casein)
Migration* Without IL-I
Incubation of PMNC with IL- 1 2 h (I I experiments) S h (three experiments) 18 h (five experiments) 0 (IL- I present during the assay; three experiments)
With IL-1
45 min
60 min
45 min
60 min
Pt
86-3 +9 3 87-3 + 5 2 67 6+ 13 0 870 + 7 4
108 3 + 5 2 ND
70 9 + 6 7 715 +4 9 66 0+ 12 0 70 5 + 8 7
97 5 +±64 ND ND ND
< 0 0005
ND ND
0 1 < 0.05
* Migration (,um) into a 3-pm pore filter during a 45-min assay (see Materials and Methods). t Significance between leucocyte locomotion without and with previous incubation of PMNC with IL- 1; comparison
between identical durations of assays. ND, not done.
Table 2. Inhibition of leucocyte locomotion by various concentrations of ultrapure IL-1
Table 3. Inhibition of leucocyte locomotion by ultrapure IL-1, rIL-la and rIL-lf,
IL- I concentration (U/ml)
(U/ml)
0 02 1 2 10
Distance of migration (pm)
P*
837+34 71 1+29 670+10 65 7+4 3 676+10