Inhibition of polymorphonuclear leukocyte chemotaxis by streptokinase-streptodornase

Leukotaxis of circulating polymorphonuclear leukocytes (PMN's) appears to be a critical step in the generation of local inflammatory reactions. The present studies demonstrate a reversible, dose-dependent inhibition of PMN chemotaxis by streptokinase-streptodornase (SK-SD). This inhibition does not appear to depend upon the presence of lymphocytes or the release of chemokinetic humoral factors and phagocytic and bactericidal capacities of PMN's are unaffected by the doses of SK-SD employed.     

Inhibition of some human neutrophil functions by the cyclooxygenase inhibitor ketorolac tromethamine.

Ketorolac tromethamine, a new nonsteroidal anti-inflammatory agent of the pyrrolo-pyrrole group, was assayed for inhibitory effects on polymorphonuclear leukocytes (PMN) in a variety of systems. Ketorolac inhibited PMN superoxide anion generation, lysozyme release, myeloperoxidase release, adherence to plastic surfaces, and chemotaxis in response to N-formyl-methionyl-leucyl-phenylalanine (fMLP) in a dose-dependent manner. Ketorolac also inhibited phorbol myristate acetate-stimulated adherence of PMN to bovine pulmonary artery endothelial cells. The drug inhibited lysozyme and myeloperoxidase release by PMN in response to C5a but failed to inhibit C5a stimulation of PMN in any of the other assays. Levels of ketorolac required to inhibit PMN function in most systems were in the range of 0.2 to 1.0 mg/ml, but chemotaxis to fMLP was inhibited by concentrations of ketorolac as low as 1 microgram/ml. Ketorolac, currently the only nonsteroidal anti-inflammatory drug available in a parenteral form may have therapeutic usefulness in a variety of conditions thought to be mediated in part by PMN, including sepsis.     

The effects of polyanions on NBT Reductions hexose monophosphate shunt activity, and ultrastructure of polymorphonuclear leukocytes.

Heparin causes enhanced nitroblue tetrazolium (NBT) reduction by polymorphonuclear leukocytes (PMN's). To determine the mechanism of this stimulation, samples of 1 to 3 x 10(7) PMN's were incubated with various concentrations of heparin, chondroitin sulfate A (CSA), and chondroitin sulfate B (CSB), with and without NBT. The effect of the polyanions (PA) on PMN hexose monophosphate shunt (HMPS) activity was determined by the production of 14CO2 from glucose-1-14C by the leukocytes. NBT reduction was evaluated histochemically and spectrophotometrically at 515 mmu. Samples of PMN's in heparin and heparin-NBT mixtures were examined by electron microscopy after various incubation periods. Increased NBT reductions by PMN's was found when leukocytes were incubated with heparin, CSA, and CSB, but these compounds had no effect on the HMPS activity of PMN's unless NBT was added. Electron microscopy of samples that contained heparin-NBT revealed an insoluble complex that was phagocytosed by the leukocytes. The stimulation of PMN oxidative metabolism and NBT reduction that follows incubation with PA-NBT appears to be directly related to ingestion of this particulate complex by the leukocytes.     

Innate immune mechanisms in the pathogenesis of systemic lupus erythematosus (SLE).

Immune imbalance in SLE increases the susceptibility to infectious diseases. The aim of this study was to analyze several mechanisms related to non-specific immunity in this autoimmune disorder. We studied in vivo CD11b expression, phagocytosis, and chemotaxis in polymorphonuclear cells (PMN) from SLE patients. All tests were also performed under hrIL-8 stimulating conditions and analyzed by flow cytometry. Intracellular leucocyte (monocytes and PMN) enzyme activity was evaluated using specific substrates for cathepsin B and D, collagenase, and oxidative burst by flow cytoenzymology. An exaggerated in vivo CD11b expression was observed on PMN from SLE patients without noticeably in vitro effect upon hrIL-8. Similarly both, phagocytosis and chemotaxis were diminished and showed no response to hrIL-8 stimulation. The opposite was found in PMN from controls. Intracellular enzyme activity was comparable between groups as far as cathepsin B and D are concerned. A tendency of decreased oxidative-burst induction was noted in monocytes and PMN from SLE patients, whereas collagenase activity was found clearly increased in both leucocyte subpopulations. Our results may represent a deficient ability of the innate immune mechanisms for the clearance of infectious agents, immune complexes, satisfactory resolution of inflammatory processes and tissue repair in SLE.     

Non-chemotactic influence of CXCL7 on human phagocytes. Modulation of antimicrobial activity against L. pneumophila

We have investigated the role of CXCL7 in the immune response of human phagocytes against the intracellular bacteria Mycobacterium tuberculosis and Legionella pneumophila. We have observed that polymorphonuclear neutrophil (PMN) chemotaxis induced by the supernatants of infected monocyte derived macrophages (MDM) may be attributed to CXCL8 rather than CXCL7, although both chemokines are present in large quantities. We have also found that CXCL7 is present not only in the supernatants of MDM, but also in the supernatants of PMN of some, but not all, individuals. Western blot analysis revealed that, in both MDM and PMN supernatants appeared two bands with molecular weights consistent with the platelet basic protein (PBP) and the neutrophil activating protein-2 (NAP-2) sizes. Regarding the influence on infected cells, recombinant NAP-2 enhanced the antimicrobial activity of IFNγ activated MDM against L. pneumophila, but not against M. tuberculosis. In addition, U937 cells transfected with a NAP-2 construct inhibited the intracellular multiplication of L. pneumophila, supporting its role in the modulation of the antimicrobial activity. Finally, U937 cells transfected with the NAP-2 construct showed an adherence that was dramatically enhanced when the substrate was fibronectin. We conclude that human phagocytes produce CXCL7 variants that may have a significant influence on the immune response against bacterial pathogens.     

Adherence and Migration of Guinea Pig Granulocytes After Enzyme Treatment of the Cell Surface

Glycogen-induced polymorphonuclear granulocytes (PMN) from the peritoneal cavity ofguinea pigs were examined (1) for their adherence to nylon fibers in the absence and presence of the adherence-enhancing chemotactic peptide formyl-methionyl-leucyl-phenylalanine (f-MLP), (2) for their random migration through the filter of a Boyden chamber and (3) for their chemotactic migration towards f-MLP. The cells were analyzed before and after treatment with the enzymes neuraminidase, papain and trypsin. PMN adhesiveness was increased by neuraminidase digestion but reduced by treatment with the proteolytic enzymes. Neuraminidase and trypsin had no effect on cell migration, while papain reduced random migration without affecting f-MLP-induced chemotaxis. The data suggest that the type of adherence measured by the nylon fiber method differs from the temporary attachment of cells migrating through a chemotaxis filter towards an attracting substance.     

Inhibition of Neutrophil Chemotaxis by a Monoclonal Antibody (TM316)

A monoclonal antibody, TM316, IgM kappa, was raised against the human monocytoid leukaemia cell line THP-1, and was shown to inhibit polymorphonuclear leucocyte (PMN) chemotaxis. The molecular weight (MW) of the protein on the PMN membrane with which TM316 bound was about 78,000. TM316 inhibited the chemotactic response of human PMN induced by at least three kinds of chemotactic factors (activated serum, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP), and a lymphocyte-derived chemotactic factor (LDCF)) to the same extent. The extent of inhibition of chemotaxis by TM316 was strongly correlated with the quantity of cellular surface antigen recognized by TM316, when a cell sorter was used for analysis. TM316 did not alter the number of Fc or complement receptors of PMN, nor did it affect luminol-enhanced chemiluminescence (CL), lysosomal enzyme release, adherence, or superoxide anion generation by PMN. TM316 seemed to recognize a common surface antigen which was necessary only for the process of chemotaxis.     

Mechanisms of tumor necrosis factor-alpha alteration of PMN adhesion and migration.

We have investigated the effects of recombinant human tumor necrosis factor-alpha (rhTNF alpha) on polymorphonuclear leukocytes (PMNs), concentrating on the mechanisms involved in the alterations of PMN-directed migration and adherence by this cytokine. RhTNF alpha profoundly suppressed PMN chemotaxis toward FMLP by 80%. At similar concentrations, it enhanced adhesion to gelatin-coated plastic dishes by more than tenfold and increased the expression of the CD11b antigen to 182% of the control. The monoclonal antibody 60.1, which is directed against the alpha chain of the CD11b/CD18 complex, completely blocked rhTNF alpha, induced inhibition of the chemotactic response to FMLP, and rhTNF alpha induced hyperadherence, suggesting that these effects were related to rhTNF alpha's effects on CD11b antigen expression. The fluid state of the PMN membrane was also decreased by rhTNF alpha. N-butanol, a known membrane fluidizer, partially inhibited the effect of rhTNF alpha on membrane fluidity and chemotaxis and completely reversed its effects on adherence and the expression of the CD11b antigen. Pentoxifylline, an agent that has previously been studied for its ability to prevent some effects of rhTNF alpha on PMNs, completely prevented the effect of rhTNF alpha on chemotaxis, the expression of the CD11b antigen, and membrane fluidity. Pentoxifylline partially prevented changes in adherence caused by this cytokine. Increased CD11b antigen expression caused by rhTNF alpha may result in enhanced PMN adhesion and suppression of migration. These events may, in turn, lead to the accumulation of PMNs on the vascular endothelium, resulting in the extensive vascular and tissue damage that is seen in gram-negative sepsis.     

PMN migration measured by bottom filter counts in modified Boyden chambers. Chemokinetic deactivation and vinblastine inhibition of chemotaxis

Polymorphonuclear leukocyte (PMN) migration and chemotaxis were studied by a modified Boyden chamber technique in 100% autologous serum with casein as attractant. PMN chemotaxis was induced by increasing the top gradient concentration of casein to 20 mg/ml. Under non-gradient condition, casein 20 mg/ml inhibited PMN migration, which was thought to be due to chemotactic deactivation. Vinblastine 0.01 microgram/ml partially inhibited PMN migration in a casein gradient with a top gradient concentration of 20 mg casein/ml. It is suggested that PMN migration in the casein gradient was composed of both vinblastine-sensitive chemotaxis and vinblastine-resistant chemotaxis.     

Effector mechanisms of intestinally induced immunity to Pseudomonas aeruginosa in the rat lung: role of neutrophils and leukotriene B4.

This paper investigates the effector mechanisms of immune clearance in the lungs of rats immunized against mucoid Pseudomonas aeruginosa. After the gut-associated lymphoid tissue was primed and after a subsequent pulmonary challenge with live bacteria, significantly accelerated bacterial clearances from the lung and raised levels of anti-P. aeruginosa antibodies in sera (immunoglobulin G [IgG], IgA, and IgM) and bronchoalveolar lavages (IgG and IgA) were observed for all immune animals. These changes were associated with enhanced recruitment, chemotaxis, chemokinesis, phagocytic indices, and chemiluminescence of pulmonary polymorphonuclear neutrophils (PMN). In the alveolar spaces of immune animals, an increase in the level of PMN recruitment was not associated with higher levels of leukotriene B4 (LTB4). In contrast, in nonimmune animals that were intratracheally infected with P. aeruginosa, the levels of recruitment and activity of alveolar PMN were lower than those in immune rats but PMN infiltration correlated with a significant increase in the synthesis of LTB4 in the alveolar space. In pulmonary tissue, LTB4 synthesis for both groups was elevated. These findings suggest that accelerated clearance of mucoid P. aeruginosa from the lungs of intestinally immunized rats is due at least in part to factors that induce the enhancement of PMN recruitment and activity in the alveolar space. The mediators that regulate this enhanced response remain unknown but do not seem to include LTB4. The high levels of LTB4 measured in the bronchoalveolar lavages and pulmonary tissues from nonimmune animals infected with live bacteria implicate LTB4 as an important amplifier of the inflammatory response during acute pulmonary infections with mucoid P. aeruginosa in unimmunized hosts.     

CD14 deficiency leads to increased MIP-2 production, CXCR2 expression, neutrophil transmigration, and early death in pneumococcal infection

CD14 is a myeloid receptor for bacterial cell membrane/wall components, for which we previously showed a strong induction in cerebrospinal fluid (CSF) during meningitis. Here, we studied CD14 function in murine Streptococcus pneumoniae meningitis by using wild-type (WT), CD14(-/-) mice, and WT mice pretreated with neutralizing anti-CD14 antibodies. Early polymorphonuclear leukocytes (PMN) immigration was more pronounced in CSF of CD14(-/-) than of WT mice. This was not a result of altered adherence molecule expression in blood and CSF PMN or brain endothelial cells. Macrophage inflammatory protein-2 (MIP-2) and keratinocyte-derived chemokine levels were similar in CSF in both strains, but MIP-2 was higher in infected brain and in brain-derived endothelial cells infected in vitro in CD14(-/-) than in WT mice. CD14(-/-) PMN demonstrated increased expression of CXC chemokine receptor 2 (CXCR2) after infection and stronger in vitro chemotaxis than WT PMN toward CSF from WT or CD14(-/-) mice and toward MIP-2. Excess PMN migration in CD14(-/-) mice did not result in improved bacterial clearing but in increased tumor necrosis factor in CSF, higher disease severity, and earlier death. Pretreatment with anti-CXCR2 reduced PMN infiltration into CSF and brain MIP-2 production and abolished earlier mortality in CD14(-/-) mice. In conclusion, CD14 plays a protective role in pneumococcal meningitis by slowing PMN migration via MIP-2 and CXCR2 modulation.     

Influence of porcine natural modified surfactant on chemotaxis and oxidative metabolism of polymorphonuclear leukocytes

Surfactant has been shown to influence a variety of immune functions. However, for in vitro studies most investigators used a single surfactant concentration often far below what has to be expected in bronchoalveolar fluid following surfactant replacement therapy. We studied the chemotactic activity and the oxygen metabolite release of human polymorphonuclear leukocytes (PMN) following incubation with Curosurf, a porcine modified natural surfactant preparation, at concentrations ranging from 1 to 16 mg/ml. In the presence of 1% bovine serum albumin, surfactant at 1 and 4 mg/ml enhanced anaphylatoxin C5a-related chemotaxis, whereas a higher dose of 16 mg/ml was inhibitory. Furthermore, Curosurf itself demonstrated a concentration-dependent chemotactic effect. Oxygen metabolite release, as measured by nitroblue tetrazolium reduction, was significantly diminished at surfactant concentrations of 8 and 16 mg/ml. This effect was most pronounced when group B streptococci at concentrations 5 x 10(9) CFU/ml were applied for PMN stimulation. We conclude that the effects of surfactant on PMN immune functions are not only concentration-dependent but also influenced by the degree of PMN stimulation.     

Hyperacute renal allograft rejection in the rabbit. The role of platelet-activating factor and of cationic proteins derived from polymorphonuclear leukocytes and from platelets

The macroscopic signs of rejection, the levels of circulating transplantation antibodies, the histologic and immunocytochemical aspects, and the levels of platelet-activating factor (PAF) present in the venous blood were studied in two groups of rabbits that had received renal allografts, one group presensitized with multiple skin grafts and a second group unsensitized, as well as in a third group of rabbits that had received renal autografts. All of the eight allografts hyperacutely rejected by presensitized recipients had deposits of rabbit IgG, IgM, and C3 along the endothelia of the vessels and massive intravascular accumulation of platelets (Pt) as soon as 5 minutes after revascularization. PAF release was detected in 2 to 10 minutes after revascularization and was present throughout most of the 60 minutes of observation. Sixty minutes after transplantation Pt and polymorphonuclear leukocytes (PMN) obliterated the vasculature and deposits of Pt- and PMN-derived cationic proteins were detected in the lumina and in the walls of the capillaries. Similar, but less severe, findings were observed in three of six renal allografts which had transient episodes of rejection after transplantation into presensitized recipients. In contrast, circulating transplantation antibodies, macroscopic signs of rejection, vascular immune deposits, release of PAF, and microvascular thrombosis were not detected in renal allografts transplanted into unsensitized recipients or in renal autografts. The results indicate that in hyperacute renal allograft rejection there is an immediate fixation of transplantation antibodies and complement of the recipient to endothelial antigens of the graft, local release of PAF, and massive accumulation, aggregation, and degranulation of Pt and PMN in the vasculature, resulting in a binding of Pt- and PMN-derived cationic proteins to the walls of the capillaries. It is conceivable that PAF release and Pt and PMN cationic proteins may contribute, together with other lysosomal enzymes, vasoconstriction, and coagulation, to the pathogenesis of antibody- and complement-mediated hyperacute graft injury.     

Use of a solid phase red blood cell adherence method for pretransfusion platelet compatibility testing

A solid phase red blood cell adherence method has been used for platelet antibody detection and crossmatching for refractory platelet recipients. Patient sera were first screened for HLA or platelet-specific antibodies, then crossmatched with potential apheresis platelet donors. The overall correlation of platelet crossmatch results with transfusion outcome was 97% in patients with no evidence of nonimmune platelet destruction. The solid phase red blood cell adherence method provided a feasible and effective alternative to HLA matching as a means of donor selection for refractory platelet recipients. The speed and simplicity of this method may allow most hospital laboratories to perform platelet antibody screening before routine platelet transfusions.     

Irreversible neutrophil aggregation. A mechanism of decreased newborn neutrophil chemotactic response.

To investigate the neutrophil-neutrophil interactions of the newborn for possible clues to the etiology of decreased newborn neutrophil (PMN) chemotaxis, the authors compared adult and newborn C5a-induced PMN aggregation and chemotaxis at various PMN concentrations. Using Craddock's technique of C5a-induced aggregation, the authors found that the newborn lacks the normal biphasic aggregation-deaggregation seen in the adult, suggesting irreversible aggregation similar to that seen when adult PMNs are pretreated with cytochalasin-B. Chemotaxis of adult and newborn PMNs was studied with a modified Gallin radiolabel technique. A linear correlation between PMN concentration and corrected chemotactic response was found with both adult (r2 = 0.93) and newborn (r2 = 0.90) PMNs in the range 0.1 X 10(6) to 20 X 10(6) PMNs/ml. Random migration was not augmented by increased PMN concentration. The augmentation of newborn PMN chemotaxis was less than that of the adult (adult slope = 2426; newborn slope = 983). Irreversible newborn PMN aggregation may be the underlying event producing decreased PMN chemotaxis and interfering with the normal chemotactic augmentation caused by increased PMN concentration.     

Modulation of human polymorphonuclear leukocyte chemotaxis by leukocyte extracts,bacterial products,inflammatory exudates,and polyelectrolytes

Human polymorphonuclear leukocytes (PMN) chemotaxis was tested during exposure to leukocyte and platelet extracts, a variety of polyelectrolytes, inflammatory exudates, and bacterial products. The chemoattractants employed were either zymosan-activated serum or supernatant from autolyzedStaphylococcus aureus. Chemotaxis to both chemoattractants was markedly inhibited by leukocyte and platelet extracts; inflammatory exudates; anionic polyelectrolytes, DNA, hyaluronic acid, liquoid; and by cationic polyelectrolytes, histone, protamine base, protamine sulfate, and myeloperoxidase. Inhibition was also found with elastase, collagenase, pepstatin, and epsilon-aminocaproic acid. Bacterial products, such as lipoteichoic acid and lipopolysaccharides, and extracts of human dental plaque inhibited chemotaxis. No inhibition of chemotaxis was observed with heparin (<10 g/ml), chondroitin sulfate, phosphatidylethanolamine and phospatidylserine. Indeed, chondroitin sulfate markedly enhanced chemotaxis and antagonized the inhibitory effect of leukocyte or platelet extract. None of the agents employed was toxic to PMN as judged by trypan blue exclusion. These observations suggest that cationic polyelectrolytes and inflammatory exudates influence PMN surfaces, modifying interaction with chemoattractants. Assessment of the role of PMN chemotaxis inThis investigation was supported by research grants obtained through the courtesy of Dr. Samuel Robin of Cleveland, Ohio, the Alpha Omega Chapter, Friends of the Hebrew University in Manchester, England, and grant AI 08821 obtained from the National Institutes of Health for Dr. Paul G. Quie. Part of this investigation was performed at the Department of Pathology, School of Dental Medicine, University of Pennsylvania, Philadelphia supported by a research grants DE-02523, and DE-03995 from N.I.D.R.Dr. Isaac Ginsburg was the Lasby Visiting Professor at the University of Minnesota June–September, 1979, and a Visiting Professor at the University of Pennsylvania, October–December, 1979.Dr. Paul G. Quie is the American Legion Heart Research Professor.     

Non-specific immunity in aging: deficiency of monocyte and polymorphonuclear cell-mediated functions

Peripheral blood monocytes obtained from 55 aged donors were evaluated for their chemotactic and phagocytic capacity. In the same subjects, polymorphonuclear cell-mediated functions were studied by chemotaxis, phagocytosis, nylon fiber adherence and nitroblue-tetrazolium reduction assay. Monocytes showed a normal chemotactic responsiveness to zymosan-activated serum, while the chemotactic activity induced by leukocyte-derived chemotactic factor and phagocytosis were rather depressed. A dramatic impairment of polymorphonuclear cell-mediated immune response was also observed. In fact, in spite of a normal nylon fiber adherence, chemotaxis, phagocytosis and nitroblue-tetrazolium reduction capacity were significantly depressed by the aging process. These data suggest that the deficiency of non-specific immunity may play an important role in the increased susceptibility to infections in aged donors.     

Effects of the two varieties of Cryptococcus neoformans cells and culture filtrate antigens on neutrophil locomotion.

Cryptococcus neoformans var. gattii (serotype B and C) isolates have a relative predilection for immunocompetent hosts, and C. neoformans var. neoformans (serotype A and D) isolates have a relative predilection for immunocompromised hosts, suggesting that normal host resistance to the former may be relatively inefficient compared with that to the latter variety. In order to assess the possibility that normal cellular host defense is inadequate in protecting against C. neoformans var. gattii, we compared the two varieties of C. neoformans cells and their culture filtrate antigens (CneF) with respect to effects on neutrophil (polymorphonuclear leukocyte [PMN]) locomotion. In a 48-well modified Boyden chamber, the cells and CneF of C. neoformans var. neoformans (serotype A and D) isolates stimulated chemotaxis and chemokinesis of human PMN and activated a complement component(s) in pooled human serum to become a chemoattractant(s) for human PMN. In contrast, the cells and CneF of C. neoformans var. gattii (serotype B and C) isolates did not stimulate chemotaxis or chemokinesis in human PMN but rather inhibited chemokinesis and chemotactic responses of PMN to pooled human serum and formylmethionyl leucyl phenylalanine. Neither of the CneF from the C. neoformans var. gattii isolates was cytotoxic to PMN. Furthermore, with the mouse model, we found that CneF from C. neoformans var. neoformans caused migration of PMN into gelatin sponges implanted in naive and immunized mice, whereas CneF from C. neoformans var. gattii inhibited PMN migration into sponges. Our results, combined with findings of others showing reduced PMN infiltration in lungs of mice infected with C. neoformans var. gattii compared with PMN infiltration in lungs of mice infected with C. neoformans var. neoformans, indicate that the relative inadequacy of normal host resistance mechanisms to prevent infection with C. neoformans var. gattii results, in part, from inhibition of PMN migration to the site of the organism.     

Allergen-induced depression of neutrophil chemotaxis in allergic individuals

In previous studies we have shown that neutrophil (PMN) chemotaxis is depressed in patients with allergic disease, hyperimmunoglobulinemia E, and recurrent infections. Although a number of these patients have now been described, little is known about the mechanism of defective PMN function in such individuals. The present studies were carried out to define the relationship between defective PMN chemotaxis and allergic hypersensitivity. Neutrophil chemotaxis was assessed in individuals with sensitivity to ragweed pollen as documented by intradermal skin testing. In each patient, chemotaxis was measured before and after in vitro exposure of their leukocytes to ragweed. A total of 21 individuals in 3 groups have been studied. These include 7 patients with allergic symptomatology and recurrent infections who were ragweed sensitive, 6 patients with a prior history of ragweed sensitivity who had undergone successful desensitization, and 8 nonatopic control subjects. Following in vitro exposure of the leukocytes of the 7 symptomatic patients to ragweed, a profound depression (54% ± 16%) in chemotatic activity was noted. In contrast the control subjects and 6 desensitized patients showed no depression of PMN chemotaxis. Further studies were carried out which demonstrate the antigen-specific nature of the phenomenon and the ability to transfer the depression to normal cells suspended in allergic serum. These data suggest that defective PMN chemotaxis in allergic patients results from an interaction, either directly or indirectly, of antigen with sensitized leukocytes. Specific immunotherapy may have an effect to prevent the chemotactic abnormality in these patients.     

The adherence of polymorphonuclear leucocytes to an albumin-coated glass surface. Effects of therapeutic concentrations of the Catharantus derivatives vincristine, vinblastine and vindesine

The Catharantus derivatives are microtubule antagonists employed in immunosuppression and chemotherapy of neoplasms. The role of cytoplasmic microtubules in polymorphonuclear leucocyte (PMN) adherence was studied by means of therapeutic concentrations of the Catharantus derivatives vincristine, vinblastine and vindesine. PMN adherence was measured as retention on an albumin-coated glass surface. PMN adherence was reduced by 4-54% by the Catharantus derivatives, as compared with control values. The suppression of adherence was statistically significant. Since the Catharantus derivatives are microtubule antagonists, it is reasonable to assume that PMN adherence is a partially microtubule-dependent process. It is suggested that reduction of PMN adherence could account for at least part of the immunosuppressive properties of the Catharantus derivatives.