Differential recognition of microfilarial chitinase, a transmission-blocking vaccine candidate antigen, by sera from patients with Brugian and Bancroftian filariasis.

We examined the reactivity of human sera with recombinant microfilarial chitinase and with the antigenic determinant on the native parasite molecule identified by monoclonal antibody (MAb) MF1. In Brugian filariasis, the MF1 epitope is preferentially recognized by residents of endemic areas who remain amicrofilaremic and asymptomatic despite lifelong exposure to filarial worms. Reactivity with filarial chitinase and its MF1 epitope inversely correlates with microfilaremia levels in Bancroftian filariasis and is associated with a prolonged amicrofilaremic state following a single course of treatment with diethylcarbamazine. Chitinase does not appear to be a target of human antibodies that promote the adherence of cells to microfilariae, even though MAb MF1 itself promotes antibody-dependent, cell-mediated cytotoxic (ADCC) reactions that kill microfilariae in vitro. Such ADCC reactions are most often mediated by sera from amicrofilaremic patients with chronic elephantiasis that contain low or undetectable levels of IgG antibodies to chitinase. In contrast, antibodies to the MF1 epitope on this microfilarial stage-specific antigen are mostly present in amicrofilaremic donors without clinical lymphatic disease. These observations indicate that antibodies to the MF1 epitope of microfilarial chitinase reflect some degree of immune resistance to microfilaremia in a subgroup of patients with asymptomatic lymphatic filariasis. The amicrofilaremic state of individuals with chronic lymphatic disease appears to be mediated by reactivity to a different parasite antigen(s).

Evaluation of a recombinant parasite antigen for the diagnosis of lymphatic filariasis.

We evaluated the usefulness of a recombinant parasite antigen (recSXP1) for the serologic diagnosis of lymphatic filariasis. A large proportion of sera from microfilaremic donors living in five different endemic countries (356 of 446 [80%]) contained IgG antibodies to recSXP1, as do sera from approximately 33% of amicrofilaremic patients with acute filarial disease and/or indirect evidence of active filarial infection. Exposure to filarial worms per se does not appear sufficient to elicit an anti-SXP1 antibody response. Thus, this serologic test identifies a large proportion of persons with active lymphatic filariasis among residents of endemic areas.

Differential recognition of microfilarial antigens by sera from immigrants into an area endemic for brugian filariasis.

Studies in animal models indicate that antibodies to surface antigens of microfilariae participate in the control of parasitaemia resulting from infections with lymphatic filarial nematodes. In an attempt to identify parasite antigens that elicit such 'protective' host responses, we compared the antigen recognition patterns of persons who remained amicrofilaraemic after 3-6 years of exposure to Brugia malayi with those of individuals who developed patent filariasis during the same period. IgG antibodies in sera from immigrants identified between 0 and 25 microfilarial antigens on Western blots. The highest degree of reactivity was observed with antigens in the 65-75 kD and 20-30 kD ranges, and with a group of high mol. wt antigens (greater than 180 kD). Sera from amicrofilaraemic donors preferentially reacted with 70/75 kD microfilarial antigens. A proportion of such sera inhibited the binding of monoclonal antibody MF1 to its target epitope; eight of nine inhibitory sera were from patients with active infections, evidenced by the presence of microfilariae or filarial antigens in the donors' blood, but who were amicrofilaraemic. These results indicate that some amicrofilaraemic residents of areas where brugian filariasis is endemic develop immune reactions to a microfilarial stage-specific antigen that was previously identified as a potentially 'protective' parasite antigen in animal models of lymphatic filariasis.

Immune response of humans to the circumsporozoite protein of Plasmodium falciparum: limited T cell response to the immunodominant central repeat region.

Most adults in highly malarious areas have antibodies to the repeat region of the circumsporozoite protein of Plasmodium falciparum. To determine if a T cell epitope on the repeat region stimulated T cell help for this antibody, we used R32tet32, a recombinant construct derived from the repeat region of the circumsporozoite protein of P. falciparum, to stimulate in vitro mononuclear cells from residents of an area hyperendemic for malaria. Three groups differing in the length of time they had resided in a malarious area were studied. The percentage of individuals in each group who had positive antibody responses to R32tet32 increased with increased exposure to malaria. However, antibody positivity was not correlated with in vitro lymphocyte proliferation responses to the antigen. Lymphocytes from 79% of the individuals showing serum antibodies to R32tet32 failed to respond in a lymphocyte transformation assay, suggesting that T cell helper activity in these individuals was based upon the recognition of a T cell epitope not located within this peptide.

Regulation of immune responses in lymphatic filariasis.

The nature and intensity of immune reactions to filarial antigens appear to be controlled by two broad mechanisms: immunoregulation and immune tolerance. Parasite molecules of high molecular weight activate suppressor T lymphocytes; suppressive parasite products are present in sera from microfilaraemic patients. Prenatal or perinatal exposure to soluble parasite antigens may influence a person's future ability to react to filarial antigens.

In vitro growth inhibition of Plasmodium falciparum by sera from tropical splenomegaly syndrome patients.

Sera from tropical splenomegaly syndrome (TSS) and non-TSS patients from the same village were examined for their ability to inhibit the in vitro growth of Plasmodium falciparum. Using synchronized malaria cultures, sera from both groups inhibited parasite development only if added before merozoite reinvasion of erythrocytes had occurred. There was no significant difference in the degree or apparent mechanism of inhibition caused by TSS and non-TSS sera. These results suggest that the aberrant immune response that results in TSS may not be associated with the elaboration of unique serum factors that differentially inhibit growth of the parasite in vitro.

Identification of Brugia malayi in vectors with a species-specific DNA probe.

We evaluated the potential value of a cloned sequence of genomic DNA of Brugia malayi as a species-specific probe. Clone pBm 15 reacted with all stages of 8 different geographic isolates of B. malayi and cross-hybridized with microfilariae of B. timori. It did not hybridize with Wuchereria bancrofti or with B. pahangi, W. kalimantani, Dirofilaria repens, Breinlia booliati or Cardiofilaria species, animal filariids that can be sympatric with B. malayi. P32-labeled clone pBm 15 correctly identified mosquitoes infected even with 1 infective larva of B. malayi. This specific DNA probe should be an invaluable tool to monitor control programs of Brugian filariasis.

Antibody-mediated killing of suppressor T lymphocytes as a possible cause of macroglobulinemia in the tropical splenomegaly syndrome.

To investigate the pathogenesis of macroglobulinemia in the tropical splenomegaly syndrome (TSS), we assessed the functional activity of B lymphocytes and T cell subsets in a pokeweed mitogen-driven assay of immunoglobulin synthesis. Mononuclear cells from patients with TSS produced more IgM than cells from village or from distant controls. This appeared to result from a decrease in the number and/or activity of suppressor T cells of the T8+ phenotype. The lack of functional suppressor T lymphocytes was associated with the presence in sera from patients with TSS of IgM antibodies that specifically killed T8+, 9.3-, 60.1+ T cells from normal donors. These results support the hypothesis that macroglobulinemia in TSS results from defective immunoregulatory control of B cell function, and that this may be caused by lysis of suppressor T cells by specific lymphocytotoxic antibodies produced by patients with this syndrome.

Human intestinal parasitism in three areas of Indonesia: a survey.

Stools from 1387 people were examined quantitatively for eggs of nematode parasites. The people were residents of Padangganting, near Sawahlunto, Sumatra (227 people), Sukamaju and Cibungur rubber plantations near Sukabumi, Java (831 people) and the villages of Mahima and Rabo near Reo, Manggarai, Flores (329 people). Ascaris lumbricoides, Trichuris trichiura and Necator americanus were common; Ancylostoma was not observed. Prevalence of parasitism was highest in Java and lowest in Flores. Egg counts were low, suggesting low worm burdens; intensity of infection with Ascaris and Trichuris was highest in Java, while hookworm was highest in Sumatra. Ascaris infections decreased and hookworm infections increased in prevalence and intensity with age; Trichuris was unaffected. A sex-related difference was observed only in Sumatra, where more females than males were infected with Ascaris. Village-to-village variation in prevalence of nematode infestation was observed in Flores and on the Sukamaju plantation; the topography of other areas did not allow for village comparisons. Ad hoc anthelmintic treatment of the residents of Cibungur plantation was reflected in reduced parasitism by Ascaris and Trichuris, but not hookworm, compared with neighbouring Sukamaju. Comparison of the patterns of disease in the three areas, in terms of the occurrence of single, double or triple infections, revealed marked differences. In Java most people with parasitism had triple infections. In Sumatra the most common expression of parasitism was single infection with hookworm. In Flores single infection with Ascaris prevailed. It is suggested that the clinical effects of intestinal parasitism might depend on the pattern of infection. Samples collected in Flores were also examined for protozoa which were found in about half the population; Entamoeba coli and E. histolytica were most common.

Reduction of suppressor T lymphocytes in the tropical splenomegaly syndrome.

To study the pathogenesis of tropical splenomegaly syndrome, we compared immunologic findings in patients from Flores, Indonesia, with those obtained in local residents without splenomegaly and in controls. Villagers with tropical splenomegaly syndrome had markedly elevated levels of total IgM, higher titers of IgM antibodies to Plasmodium vivax, and reduced levels of circulating T lymphocytes. The latter were caused by a decrease in the total number of T cells with the suppressor/cytotoxic phenotype (T8+). Levels of B lymphocytes were similar in all groups. All immunologic abnormalities reverted toward normal in patients treated weekly for 9 to 26 months with chloroquine phosphate. These findings suggest that overproduction of immunoglobulins in patients with tropical splenomegaly syndrome is caused by an imbalance in the normal ratio of helper: suppressor T cells that regulate B-lymphocyte function, and that this imbalance is due to a decrease in suppressor T lymphocytes.

Comparison of immunity to malaria in Sudan and Indonesia: crisis-form versus merozoite-invasion inhibition.

Immunity to falciparum malaria was compared in two populations from malarious areas of southern Sudan and Flores, Indonesia. In Sudan, splenomegaly in adults was rare and anti-plasmodium indirect fluorescent antibody (IFA) titers were low to moderate, 1:1,280 being the modal titer. Sudanese serum was profoundly inhibitory to cultured Plasmodium falciparum, reducing incorporation of radiolabeled hypoxanthine by 63-93% and severely retarding intraerythrocytic parasite development, resulting in moribund crisis-form parasites and virtually no healthy schizonts. In Flores, 64% of the serum donors had splenomegaly greater than or equal to Hackett spleen grade 4 or 5, and the modal IFA titer was 1:10,240. Sera from Indonesia did not retard intraerythrocytic parasite development, but inhibited merozoite erythrocyte invasion 22-87%. Anti-merozoite activity did not correlate with IFA titers. The differences in principal modes of anti-parasitic activity suggest that immunity to malaria in Sudan is based on cell-mediated immune mechanisms associated with crisis forms, merozoite neutralization being of secondary importance. In contrast, malaria immunity in Flores appears to be principally based on anti-merozoite antibody, which does not cause crisis forms and allows for development of reduced numbers of healthy schizonts. This less efficient mechanism may lead to a continuous low-grade parasitemia, which could explain the high specific malaria antibody titers and adult splenomegaly in Flores as compared to Sudan. This latter approach to immunity, being less efficient than the former, apparently results in chronic malaria infections with associated high Ig titers and splenomegaly.

Chloroquine resistant Plasmodium falciparum on the island of Flores, Indonesia.

The in vitro sensitivity of Plasmodium falciparum to chloroquine was studied in parasites from 45 children on the island of Flores, Indonesia. The micro in vitro culture technique and a 12-volt battery-operated field incubator were found to be well suited to the field situation encountered. Parasites from seven children (15.6%) were resistant to chloroquine in vitro: two at 8 pico-moles of chloroquine, two at 16 pico-moles, and three at 32 pico-moles. This is the first report of chloroquine resistant from Flores.

Opposing effects of filariasis and chronic malaria on immunoregulatory T lymphocytes.

The effect of parasitic infections on immunoregulatory T lymphocytes was evaluated by means of quantitative determinations of total T cell and T-cell subset populations present in peripheral blood of patients with lymphatic filariasis, with the malaria-induced tropical splenomegaly syndrome (TSS), and with both infections. Total T-cell numbers were similar to control values in patients without TSS, and reduced in donors with TSS. OKT4+/OKT8+ T-cell ratios were decreased in microfilaremic donors without TSS (0.76 +/- 0.08) and elevated in patients with TSS without microfilaremia (1.97 +/- 0.16). Patients with dual infections tended to yield results similar to uninfected control donors. Thus, filariasis and chronic malaria (TSS) have opposing effects on the normal balance of immunoregulatory T lymphocytes. In patients with TSS, the imbalance appears to result from a decrease in the absolute number of circulating T cells with suppressor/cytotoxic phenotype.