High immunoglobulin G2 (IgG2) and low IgG4 levels are associated with human resistance to Plasmodium falciparum malaria.

There is accumulating evidence for a role of immunoglobulin G (IgG) in protection against malarial infection and disease. Only IgG1 and IgG3 are considered cytophilic and protective against P. falciparum, whereas IgG2 and IgG4 were thought to be neither and even to block protective mechanisms. However, no clear pattern of association between isotypes and protection has so far emerged. We analyzed the isotypic distribution of the IgG response to conserved epitopes and P. falciparum blood-stage extract in 283 malaria-exposed individuals whose occurrence of infection and malaria attack had been monitored for about 1 year. Logistic regression analyses showed that, at the end of the season of transmission, high levels of IgG2 to RESA and to MSP2 epitopes were associated with low risk of infection. Indeed, IgG2 is able to bind FcgammaRIIA in individuals possessing the H131 allele, and we showed that 70% of the study subjects had this allele. Also, high specific IgG4 levels were associated with an enhanced risk of infection and with a high risk of malaria attack. Moreover, specific IgG2 and IgG3 levels, as well as the IgG2/IgG4 and IgG3/IgG4 ratios, increased with the age of subjects, in parallel with the protection against infection and disease. IgG4 likely competes with cytophilic antibodies for antigen recognition and may therefore block cytotoxicity mediated by antibody-activated effector cells. In conclusion, these results favor a protective role of IgG3 and IgG2, which may activate effector cells through FcgammaRIIA, and provide evidence for a blocking role of IgG4 in malarial infection and disease.

[Analysis of the genetic factors controlling malarial infection in man]. / Analyse des facteurs génétiques contrôlant l'infection palustre chez l'homme.

Genetic factors have clearly been shown to play a role in controlling malarial infection in animal models. There is now also increasing evidence for the genetic control of malaria in man. We carried out a segregation analysis based on blood parasite load phenotype for a population of the town of Bobo-Dioulasso (Burkina-Faso). This analysis demonstrated a strong genetic effect. Our results were not consistent with the segregation of a major gene and thus suggest that parasite load is under the control of minor genes. The genetic effect was stronger in children than in adults. We carried out a regression analysis in children and found that there was an association between the phenotype for blood parasite load and the q31-33 region of chromosome 5. We identified a gene in this region, Pfil1 (Plasmodium falciparum infection levels 1), which accounted for almost 50% of the variance in blood parasite load and which played a fundamental role in the control of infection. The 5q31-33 region contains several genes encoding cytokines that regulate T lymphocytes. The identification of genes controlling malarial infection opens up new possibilities for preventive and treatment strategies. It should be possible in the near future to identify individuals at risk of malaria, who would derive the greatest benefit from preventive and therapeutic measures. Finally, a deeper understanding of these genes controlling protective immune responses could be of value for the development of vaccines.

Genetic dissection of Plasmodium falciparum blood infection levels and other complex traits related to human malaria infection.

There is accumulating evidence of host genetic control in malaria infection and, in humans, some genes have been associated with severe malaria. Nevertheless, other important genes controlling blood infection levels, malarial disease and immune responses are likely to be identified. In this paper, we focus on segregation and linkage analyses of blood infection levels in an urban population living in Burkina Faso. We found evidence of a complex genetic control and a linkage to chromosome 5q31-q33. The identification of genes controlling complex traits related to malaria infection should be helpful in understanding protective mechanisms and the relationship between infection, malaria attacks and severe malaria.

Human malaria: segregation analysis of blood infection levels in a suburban area and a rural area in Burkina Faso.

The genetic control of blood infection levels in human malaria remains unclear. Case control studies have not demonstrated a strong association between candidate genes and blood parasite densities as opposed to surveys that have focused on severe malaria. As an alternative approach, we used segregation analyses to determine the genetic control of blood parasitemia. We surveyed 509 residents (53 pedigrees) in a rural area and 389 residents (41 pedigrees) in an urban area during 18 months. Each family was visited 20 times and 28 times in the urban area and in the rural area; the mean number of parasitemia measurements per subject was 12.1 in the town and 14.9 in the village. The intensity of transmission of Plasmodium falciparum was 8-fold higher in the rural area than in the urban area. Using the class D regressive model for both populations, we found that blood parasite densities were correlated between sibs. We obtained strong evidence for a major effect, but we found that the transmission of this major effect was not compatible with a simple Mendelian model, suggesting a more complex mode of inheritance. Moreover, there was a strong interaction between major effect and age, suggesting that the influence of the putative major gene may be more prominent in children than in adults. Further nonparametric linkage studies, such as sib pair analysis, that focus on children would help us better understand the genetic control of blood infection levels.

Malaria in humans: Plasmodium falciparum blood infection levels are linked to chromosome 5q31-q33.

Plasmodium falciparum malaria remains a major cause of morbidity and mortality in many tropical countries, especially those in sub-Saharan Africa. Human genetic control of malaria infection is poorly understood; in particular, genes controlling P. falciparum blood infection levels remain to be identified. We recently evidenced the existence of complex genetic factors controlling blood infection levels in an urban population living in Burkina Faso. We performed, on 153 sibs from 34 families, sib-pair linkage analyses between blood infection levels and chromosome 5q31-q33, which contains numerous candidate genes encoding immunological molecules. Our results, obtained by means of the two-point Haseman-Elston (HE) method and a nonparametric (NP) approach, show linkage of parasitemia to D5S393 (P=.002) and D5S658 (P=.0004). Multipoint analyses confirmed linkage, with a peak close to D5S658 (P=.0013 and P=.0007 with the HE and NP methods, respectively). The heritability of the locus was .48, according to the two-point results, and .43, according to the multipoint results; this indicates that its variation accounted for approximately 45% of the variance of blood infection levels and that the locus plays a central role in the control of parasitemia. The identification of the gene is, therefore, of major interest in understanding the mechanisms controlling P. falciparum parasitemia.

Trypanosoma brucei: polyamine oxidase mediated trypanolytic activity in the serum of naturally resistant cattle.

Trypanosoma brucei brucei are lysed when incubated in vitro in a mixture of bovine serum and polyamine. Normal bovine serum alone or polyamine alone does not show any trypanocidal activity. The bovine serum in the mixture can be replaced by purified polyamine oxidase, and addition of polyamine oxidase inhibitors blocks trypanolysis. Using this in vitro lysis test, it is shown that West African cattle which are resistant naturally to trypanosomiasis have a higher trypanolytic activity in their serum than do trypanosensitive cattle (P less than 10(-5]. Seric trypanolytic activity of individual animals remains stable when tested over a period of 18 months; moreover, it is not modified by trypanosome infection. Higher levels of seric polyamine oxidase in resistant cattle were demonstrated also by enzymatic analysis. The factors responsible for trypanolysis have been analyzed. Oxidation of spermidine by polyamine oxidase leads to the production of unstable aldehydes, acrolein, ammonia, O2-, HO, and H2O2. Acrolein and H2O2 show strong trypanolytic activity while the other products do not appear to be toxic for trypanosomes. The physiological importance of polyamine oxidase mediated trypanolysis is unclear; even at peak parasitemia in cattle (10(7) organisms/ml) it can be calculated that trypanosomes would not release enough spermidine for the generation of sufficient quantities of toxic degradation products. Additional polyamines could be released in serum from tissues damaged as a result of the infection.

High density lipoprotein levels in the serum of trypanosensitive and trypanoresistant cattle. Changes during Trypanosoma congolense infection.

Nonpermissiveness to trypanosome infection has been correlated in some instances with the presence of toxic serum factors, e.g. high density lipoproteins (HDL) of human serum can lyse T.b. brucei. The present study examines the possibility of a role for such factors in West African cattle that are resistant to trypanosomiasis. Cattle used in this study were previously selected as resistant or sensitive to trypanosomiasis under heavy natural Glossina challenge. - A comparison of the direct effect of serum from trypanoresistant and trypanosensitive Baoulé cattle on the development of pathogenic bloodstream or metacyclic forms of T. congolense, using modifications of the blood infectivity incubation test, failed to demonstrate a difference between these cattle. High density lipoproteins and cholesterol levels were compared in 115 cattle of known sensibility to trypanosomiasis. HDL-cholesterol formed 91% of the total plasma cholesterol. HDL-cholesterol levels in Zebu (mean of 111.8 mg/100 ml) were significantly higher than those in Baoulé cattle (86.2 mg/100 ml). There was no significant difference, however, in these levels between trypanoresistant (73.4 mg/100 ml) and trypanosensitive (84.5 mg/100 ml) Baoulé. Alterations in HDL-cholesterol levels were monitored during an experimental cyclic infection with T. congolense in 5 Zebu and 9 Baoulé. HDL-cholesterol levels decreased in all animals concomitantly with the appearance of trypanosomes in the blood and returned rapidly to their starting values after parasite elimination following drug treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Correlation of serum zinc levels with resistance of cattle to trypanosomiasis.

Zinc, copper and magnesium levels were determined by atomic absorption spectrophotometry in the serum of 32 cattle (Zebus and Baoulés) which were proven to be sensitive to African trypanosomiasis under field challenge and 45 cattle (Baoulés and Ndamas/Baoulés) which were proven to be resistant. Copper and magnesium levels were similar in all animals but zinc levels were higher in sensitive animals (1.50 ppm) than in resistant ones (1.10 ppm) (p less than 10(-5)); the reported normal levels of serum zinc is 1.00 ppm. These differences persisted on repeated measurements and whether individuals were infected with trypanosomes or not. Elevated levels of zinc depressed the stimulation of bovine T cells by trypanosomes in vitro and is reported to inhibit antigen presentation by macrophages. Zinc levels may be an influencial factor determining susceptibility or resistance of West African cattle to trypanosomiasis.

High and low responsiveness of bovine lymphocytes to Trypanosoma brucei in vitro: lack of correlation with resistance to trypanosomiasis.

Bovine peripheral blood lymphocytes (PBL) were stimulated to proliferate in vitro by live, irradiated or freeze-thawed Trypanosoma brucei, but not by the isolated variant surface glycoprotein. The optimal dose was 10(5) trypanosomes per 5 X 10(5) lymphocytes in 0.2 ml. Maximal proliferation was at day 5. Of the 98 cattle tested, 36 were high-responders (stimulation indexes 20-104), 49 were low or non-responders (SI 1-10) and 13 were intermediate. The responder status of individual animals did not change over a period of 1 year, nor did it alter following deliberate trypanosome infection. The stimulation was dependent on macrophage/monocyte type accessory cells, and this co-operation did not seem to be MHC restricted. Lack of stimulation of non-responder PBL did not appear to be due to the activation of suppressor cells. Accessory cells from non-responder animals could complement PBL from responders, but accessory cells from responders could not complement non-responder PBL. Responsiveness is therefore a characteristic of lymphocytes. Analysis of the surface markers of these lymphocytes or the blast cells generated in culture showed that they were a subpopulation of T cells, possibly TH cells. Analysis of PBL from 98 animals, which had been selected for trypanoresistance or trypanosensitivity under natural tsetse fly challenge, failed to establish a correlation between resistance and level of lymphocyte stimulation by trypanosomes in vitro.