T cells play a crucial role in antibodymediated and antibody-independent immunity against Plasmodium falciparum malaria. Therefore, a vaccine immunogen should include parasite-derived B-and T-cell epitopes capable of giving rise to protective responses in both systems. The P.falciparum antigen PfI55/ring-infected erythrocyte surface antigen (RESA), a vaccine candidate, contains immunodominant T-and B-cell epitopes located in the central (5') and C-terminal (3') invariant repeat regions of the molecule. To relate Pfl55/RESA-peptide-specific responses of T cells to function, T cells from P. fakciparum immune donors were activated with peptides corresponding to these immunodominant regions. Activation was measured as induction of interferon-y secretion, T-cell proliferation (DNA synthesis), or transcription and translation of interleukin 4 (IL-4) mRNA.Peptides from both regions were shown to induce interferon-y, IL-4, proliferation, or any combination. In individual donors, there was no correlation between these different activities. Rather, they were negatively correlated, demonstrating the importance of examining multiple parameters of T-cell activation when estimating the proportion of individuals responding to a given epitope. However, IL-4 mRNA and intracellular IL-4 could be induced in T cells of donors who had elevated concentrations ofserum antibodies to the same peptide that was used for T-cell activation. These results suggest that a causal relationship exists between the activation of IL-4-producing T-cell subsets and production of the anti-Pf155/RESA-specific antibodies in individuals in which immunity has been induced by natural infection. This finding has implications that should be considered for the selection of immunogens to be included in a future P. falciparum subunit vaccine and for vaccine development in general.

In mice, immune responses to subunits of defined malaria antigens are regulated by genes mapping within the MHC and it has been suggested that such genetic restriction will be a major obstacle in the development of a human malaria vaccine. The relationship between class II human leukocyte antigen (HLA) genes and immune recognition of three candidate antigens for a vaccine against Plasmodium falciparum malaria has been investigated in a human population living in a malaria endemic area of West Africa. The study population was shown to be extremely heterogeneous for HLA class II alleles and marked differences in allelic frequency were detected between members of different ethnic groups. One class II DQA-DQB combination (serological specificity DQw2) was particularly common among members of the Fula ethnic group. This haplotype was significantly associated with higher than average levels of antibody to a peptide epitope, (EENV)6, of the malaria antigen Pf155/RESA. There was little evidence of association between HLA class II genotype and cellular proliferative or interferon gamma responses to the antigens tested. Overall, the number of significant associations between immune responses and specific HLA class II haplotypes was greater than would be expected by chance but less than would be expected if class II-dependent genetic restriction were a major factor governing human immune responses to malaria antigens. Thus, although some qualitative variation in the immune response to vaccine antigens may occur in ethnically different target populations, widespread HLA-associated nonresponsiveness to a multivalent subunit malaria vaccine is unlikely.

SUMMARYThe reasons why only a small proportion of African children itifecled with Plasmodium falciparum develop severe or fatal malaria are not known. One possible reason is that children who develop severe disease have had less previous exposure to malaria itifectioti. and heticc have less acquired immunity, than ehildren who develop a mild elinical attack. To investigate this possibility we have trtcasured litres ofa wide range of anti-P. falciparum antibodies in plasma samples obtained from children with severe malaria, children with mild malaria and from children with other illnesses. Mean antibody levels in palienls with malaria were higher than those in patienis wilh other eonditions bul. with only one exception, there were no signifieant diiTerences in anlibody litres between cases of severe or mild malaria. A parasitized-erythroeyle agglutination assay was used to estimate the diversity of parasite isolates to which ehildren had been exposed; plasma samples obtained from ehildren with eerebral malaria reeognized as many isolates as did samples obtained from children with mild disease. Our findings do not provide any support for the view thai the developmetit vi\' severe malaria in a small proportion of African children infected with P. falciparum is due lo laek of previous exposure lo the infection.

The 230 kD gamete surface protein of the malaria parasite Plasmodium falciparum (Pfs 230) is a target of transmission blocking antibodies. Anti-Pfs 230 antibodies are inducedfollowing natural infection with malaria but are not ,found in all P. falciparum-exposed individuals. In this study we have shown that approximately 40% of malaria-exposed Gambians do not make antibodies to the native Pfs 230 molecule. This phenotype is remarkably stable over time and does not appear to be d a t e d to age, malaria exposure or major histocompatibility complex genotype. Comparison of antibody responses in twins indicates that the anti-Pfs 230 response is not strictly genetically controlled, but a high degree of concordance within both dizygous and monozygous twin pairs suggests that factors associated with exposure to malaria in childhood may be important in determining the subsequent immune response.

Galectins are evolutionarily conserved glycan-binding proteins with pleiotropic roles in innate and adaptive immune responses. Galectin-3 has been implicated in several immunological processes as well as in pathogen recognition through specific binding to glycosylated receptors on the surface of host cells or microorganisms. In spite of considerable evidence supporting a role for galectin-3 in host-pathogen interactions, the relevance of this lectin in the regulation of the host defence mechanisms in vivo is poorly understood. In this study, we analysed the impact of galectin-3 deficiency during infection with three distinct species of rodent malaria parasites, Plasmodium yoelii 17XNL, Plasmodium berghei ANKA and Plasmodium chabaudi AS. We found that galectin-3 deficiency showed a marginal effect on the course of parasitaemia during P. chabaudi infection, but did not alter the course of parasitaemia during P. berghei infection. However, lack of galectin-3 significantly reduced P. yoelii parasitaemia. This reduced parasitaemia in Lgals3(-/-) mice was consistent with higher titres of anti-P. yoelii MSP1(19) IgG2b isotype antibodies when compared with their wild-type counterparts. Our results reflect the complexity and singularity of host-pathogen interactions, indicating a species-specific role of endogenous galectin-3 in the control of parasite infections and the modulation of antibody responses.

Infection with Plasmodium'falciparum induces marked disturbances in normal immunoregulatory functions. Antigen-specific immunosuppression is a feature of acute malaria and has been linked to activation of CD8+ T suppressor cells. Among immune adults, cell-mediated immune responses to malaria antigens are extremely variable when measured in vitro, and there is no obvious relation between responsiveness and resistance to clinical disease. In this study, when CD8+ cells were removed from peripheral blood mononuclear cell preparations obtained from individuals who responded poorly to a soluble malaria antigen preparation, both lymphoproliferation and gamma interferon production were significantly enhanced, but responses to other soluble antigens and mitogen were unaffected. No effect of CD8+ cell depletion was seen in individuals whose undepleted mononuclear cells gave a high response to the malaria antigen. This suggests that for some malaria-exposed individuals, CD8+ cells activated in vitro by exposure to malaria antigens suppress other cellular responses and may obscure the presence of potentially protective immune mechanisms.

Objective: To determine the natural human humoral immune responses to the 19 kilodalton carboxy terminal fragment of Plasmodium falciparum merozoite surface protein 1 (MSP1 19), a malaria candidate vaccine antigen and to determine the prevalence of MAD20 and K1 alleles of P. falciparum MSP1. Design: Community based cross-sectional study.

In-vitro lymphoproliferative responses to malaria antigens are suppressed in patients with acute Plasmodium falciparum infection. Studies with other parasitic diseases have suggested that monocyte/macrophage-derived prostaglandins may be responsible for immunosuppression. Since acute malaria infection is characteristically associated with fever it is likely that prostaglandin E production will also be enhanced in these patients. In this study, indomethacin, a cyclooxygenase inhibitor which blocks the synthesis of prostaglandins, was added to the culture medium during assays of lymphoproliferative responses to malaria antigens and other soluble proteins. Responses to several antigens were enhanced in the presence of indomethacin, indicating that prostaglandins may have a generalized immunosuppressive role in malaria-infected individuals. However, responses to malaria antigens were particularly enhanced by indomethacin, suggesting that malaria-specific T-cells are especially sensitive to the effects of prostaglandin, possibly due to prior activation in vivo by circulating malaria antigens.