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The antibiotic rifamycin SV (RSV) has been successfully used by others on the local treatment of rheumatoid arthritis (R.A.). Since polymorphonuclear leukocytes (PMNL) are involved in the synovial inflammatory process we tested the 'in vitro' effect of RSV on PMNL functions, such as locomotion and phagocytosis. PMNL locomotion was evaluated by using modified Boyden Chamber and phagocytosis was tested by the number of yeast particles ingested and by NBT reduction. The, the functions of PMNL derived from 7 R.A. patients in therapy only with non steroidal anti-inflammatory drugs were compared with those of PMNL from 14 patients with non inflammatory disease (osteoporosis and osteoarthrosis) in the same therapy and with neutrophils from healthy subjects. It was demonstrated that PMNL derived from patients with both R.A. and non inflammatory disease activated their directional locomotion towards RSV, on the contrary, cells from healthy subjects were unresponsive. Moreover, only patients with R.A. showed a defective phagocytic capacity and a depression in NBT reducing activity, when PMNL were treated with RSV. This phenomenon might be correlated with beneficial effect observed after local treatment of R.A. with RSV.

The aim of our study is to evaluate the eventual activity of the total dialysate of two uremic nephrectomized patients in recirculating dialysis and the chromatographic peak of the dialysate fractionated by Sephadex column G 15 on PMN chemotaxis. Only the total dialysate and the chromatographic peak B showed inhibition of chemotaxis. On the contrary the commercial polyamines in the same concentration range, and the other chromatographic peaks, containing polyamines too, did not revealed inhibition. Our data show, therefore, that the chemotaxis inhibition could be due to the middle-molecules present in the peak B, rather than the polyamines itself. Polyamines were determined by dansylation method, separated by thin layer chromatography and quantified by spectrofluorimeter. Chemotaxis was evaluated using the modified Boyden chamber.

Lymphocyte dialysates obtained from healthy volunteers and leukemic subject were chromatographed on Bio-Gel P-4. Partially purified fractions were assayed for neutrophil locomotion: a positive chemokinetic effect and no chemotactic activity was observed in both samples.

Non steroidal anti-inflammatory drugs, such as oxametacine, are generally used in treatment of rheumatoid disease. In an 'in vitro' experimental model, the drug efficacy was tested on leukocyte functions. Locomotion, both random and directional, phagocytic activity and superoxide production of normal and rheumatoid PMNL were tested in the presence of varying concentrations of oxometacine. Locomotion was evaluated by using modified Boyden chambers; phagocytosis was tested by number of yeast particles injested and by NBT reduction; superoxide production was assayed by reduction of ferricytochrome C. In our conditions the drug exhibited a strong anti-inflammatory effect. In fact, chemotaxis and anion production were specifically depressed in a dose-dependent way.

Treatment of human neutrophils with the two chemoattractants formyl-methyonyl-leucyl-phenylalanine and opsonized zymosan or with phorbol myristate acetate induce the production of superoxide anion (O-.2), which plays a role in the inflammatory reaction and microbicidal activity. The time-course of O-.2 release and the extent to which the anion is produced are related to the nature of the stimulating agents. Anti-inflammatory non steroid drugs, such as indomethacin and oxamethacin, exert a variable depression of the oxidative burst, which does not seem correlated to the inhibition of prostaglandin synthesis, but to the lipophilic character of the drug molecule.

PGE1 and its 8-aza-analogous were assayed "in vitro" for leukocyte functions; the natural compound was not a chemotactic agent and exhibited positive chemokinesis. On the contrary, the synthetic compound chemically related to PGE1, was a chemotactic factor and inhibited the reduction of NBT during phagocytosis.

Two tripeptides, related to the chemotactic formyl-peptide, are tested for its ability to affect random and directional locomotion and to induce chemotaxis of polymorphonuclear leukocytes. The results indicate that the methyl ester of formyl-methyonyl-leucyl-phenylalanine possesses a biological activity towards human phagocytes, including a chemotactic potency, comparable to that of unmodified peptide. Therefore, the free carboxyl group does not seem essential to generate active leukoattractant. On the contrary, the replacement of the formyl group by the butyloxycarbonyl results in a drastic loss of biological activity. Our data may indicate that the two formyl-peptides interact with the same binding site on the cell membrane.

Eighteen patients with established hereditary myeloperoxidase deficiency underwent morphological, ultrastructural, cytochemical and functional analysis in order to correlate the lack of peroxidase from phagocytes with other leucocyte activities. Cytochemical and ultrastructural findings only confirmed the peroxidase defect in neutrophil and monocyte population, whereas normal peroxidase activity was detected in eosinophil granulocytes ("Alius-Grignaschi anomaly"). Morphological analysis, as determined by both ligh and electron microscopy, showed in two patients with total MPO-deficiency a large number of neutrophils (50-60%) with nuclear abnormalities very similar to Pelger-Huet's heterozygous form (two lobed neutrophils having a typical pince-nez appearance and a nuclear chromatin coarser than that of normal PMNL). Other 2 cases displayed a 50-60% five-lobed neutrophils, as occur in congenital nuclear hypersegmentation of PMNL. These findings suggest that Alius-Grignaschi anomaly and Pelger-Huet syndrome can be found associated in the same individuals, since both these abnormalities have a genetic origin. Finally, since an impaired bactericidal and fungicidal activity was observed, no patients displayed particular susceptibility to persistent or severe infections, thus confirming the presence of MPO-independent enzymatic systems.

Casein, a phosphoprotein forming aggregates in solution, exerts chemotactic activity for human neutrophils. The protein was filtered on Sephadex G 100 to obtain fractions of homogeneous mol. weight and the column fractions were tested for chemotactic activity. The chemotactic activity was found only in the lighter peak, which was then digested by trypsin. The digested material was purified by filtration through AcA-54 and the 6.000 M.W. peptide containing the most phosphate groups and tyrosines resulted chemotactic for PMN. Most likely the phosphate groups are important in chemotactic recognition.

We studied possible effects of uremic toxins and polyamines (PAs) on organ cultures of chick embryo. We added on culture media the lyophilized of total dialysate and its chromatographic peak II obtained with chromatography with Sephadex G 15. The total dialysate and peak II showed toxicity with degeneration of the culture, whereas the free PAs, we added, did not effects.

Polymorphonuclear leucocytes play an important role in the inflammatory process, and their functions are likely to be regulated by pharmacological agents. In this paper we report the "in vitro" effect of non steroidal anti-inflammatory drugs on random and chemotactic motility of human leucocytes, and demonstrate that the two forms of movement, spontaneous and directional, are differently inhibited by low concentrations of these agents. Leucocytes of patients with recurrent inflammation are unaffected by these drugs.