Granulocyte macrophage-colony stimulation factor (GM-CSF), which is a haematopoietic cytokine generated by activated T lymphocytes and macrophages during infection, was investigated for its effects on human neutrophil-mediated killing of asexual blood forms of Plasmodium falciparum. Pretreatment of neutrophils with human recombinant-GM-CSF markedly increased the parasite killing (measured by a radiometric assay), in the presence of normal serum (containing complement), immune serum (IS), purified IgG (from IS) or heat inactivated IS. GM-CSF pretreatment also enhanced phagocytosis of the parasite by neutrophils and the expression of CR3, Fc gamma RII and Fc gamma RIII receptors. Treatment of neutrophils with a combination of GM-CSF and TNF resulted in a synergistic increase in phagocytosis and killing of the parasite. The findings suggest that GM-CSF is likely to form part of the cytokine network responsible for regulating the antiparasitic activity of the neutrophil in malaria.

The interaction between Plasodium falciparum merozoites and human neutrophils, as well as the role of cytokines, complement, and antimalarial antibody on this interaction, was examined in vitro by measuring luminol-dependent chemiluminescence and phagocytosis. Merozoites, in the presence of heat-inactivated (56°C/30 min) normal serum, had very little effect on the neutrophil chemiluminescence. This response was significantly enhanced by the addition of normal serum (containing normal complement activity). In the presence of serum or plasma containing anti-P. falciparum antibodies (IS) with no detectable complement activity, the merozoites induced a marked response characterized by an increase in initial peak rate of chemiluminescence and a sustained increased rate of chemiluminescence. However, this response was not further increased if IS containing complement activity was used. Pretreatment of neutrophils with either tumor necrosis factor alpha, lymphotoxin, or gamma interferon significantly increased the neutrophil response to IS-treated merozoites, reflected in an increased initial peak rate and sustained increased rate of chemiluminescence. The effects of cytokine treatment of neutrophils and IS opsonization of merozoites were synergistic. In association with the changes in the chemiluminescence responses, IS was shown to promote phagocytosis of merozoites by neutrophils, and this event was further increased by treating neutrophils with the cytokines. The results emphasize the importance of antibody and cytokines in neutrophil-mediated damage of P. fakciparum merozoites.

Effects of fatty acids on human neutrophil-mediated killing of Plasmodium falciparum asexual blood forms were investigated by using a quantitative radiometric assay. The results showed that the antiparasitic activity of neutrophils can be greatly increased (>threefold) by short-term treatment with fatty acids with 20 to 24 carbon atoms and at least three double bonds. In particular, the n-3 polyenoic fatty acids, eicosapentaenoic and docosahexaenoic acids, and the n-6 fatty acid, arachidonic acid, significantly enhanced neutrophil antiparasitic activity. This effect was >1.5-fold higher than that induced by an optical concentration of the known agonist cytokine tumor necrosis factor alpha (TNF-␣). At suboptimal concentrations, the combination of arachidonic acid and TNF-␣ caused a synergistic increase in neutrophil-mediated parasite killing. The fatty acid-induced effect was independent of the availability of serum opsonins but dependent on the structure of the fatty acids. The length of the carbon chain, degree of unsaturation, and availability of a free carboxyl group were important determinants of fatty acid activity. The fatty acids which increased neutrophil-mediated killing primed the enhanced superoxide radical generation of neutrophils in response to P. falciparum as detected by chemiluminescence. Scavengers of oxygen radicals significantly reduced the fatty acid-enhanced parasite killing, but cyclooxygenase and lipoxygenase inhibitors had no effect. These findings have identified a new class of immunoenhancers that could be exploited to increase resistance against Plasmodium species.

Killing of Plasmodium falciparum blood forms by the differentiated human myelomonocytic THP-1Mo cell line was studied by a radiometric assay. Results showed that parasite killing was promoted by complement, antimalarial antibody, and the cytokines tumor necrosis factor alpha and gamma interferon. Differentiated THP-1Mo appears to be a useful monocytic cell line for the study of mechanisms of immunity to Plasmodium.