concanavalin-a and Filariasis

Infection with the third-stage larvae (L3) of the filarial nematode Brugia results in a Th2-biased immune response in mice and humans. Previously we have shown that the production of interleukin 4 (IL-4) is critical for down-regulating polyclonal Th1 responses in L3-infected mice. However, the in vitro neutralization of IL-4 did not fully recover the defective polyclonal Th1 responses, nor did it result in the production of any antigen (Ag)-specific Th1 cytokines, suggesting that perhaps infection with L3 does not result in priming of Th1 cells in vivo. In this study, we analyzed the role of IL-10 and Ag-presenting cells (APCs) in the spleen as additional factors controlling the Th2 bias in infected mice. Our data show that IL-10 and APCs also contribute to the suppression of mitogen-driven Th1 responses of spleen cells from infected mice. In addition, the neutralization of IL-10 or the replacement of the resident APC population from spleen cell cultures resulted in the production of Ag-specific Th1 cytokines. Irradiated spleen cells from either L3-infected or uninfected mice were able to restore Ag-specific Th1 responses in vitro. Therefore, it appears that Brugia-reactive Th1 cells are primed following infection with L3, but are actively suppressed in vivo by a mechanism that involves IL-10 and the resident APC population, but not IL-4. These results indicate that a complex interplay of cytokines and cell populations underscores the Th2-polarized response in L3-infected mice.

Topics: Aedes; Animals; Antibodies, Monoclonal; Antigen-Presenting Cells; Antigens, Helminth; Brugia pahangi; Cells, Cultured; Concanavalin A; Filariasis; Interleukin-10; Male; Mice; Mice, Inbred BALB C; Mitogens; Neutralization Tests; Nylons; Spleen; Th1 Cells

Inhibition of Concanavalin A-induced lymphocyte proliferation was used to monitor the partial purification and characterization of suppressor molecules from microfilariae of Brugia malayi. Suppressor activity was present in high molecular weight fractions of microfilarial extracts (Mr greater than 50 kd on SDS-PAGE) and was protease-sensitive but resisted treatment with sodium periodate, indicating that it is associated with parasite proteins. Suppressor activity was released by microfilariae cultured in vitro and could be detected in peritoneal exudates of intraperitoneally-infected jirds and in lymph and sera from athymic C3H/HeN mice with patent B. malayi infections. These findings indicate that immune unresponsiveness during patent filarial infections may result from the in vivo release by microfilariae of high molecular weight proteins that suppress host immune responses.

Topics: Animals; Brugia; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Elephantiasis, Filarial; Filariasis; Gerbillinae; Humans; Immune Tolerance; Interleukin-2; Lymphocyte Activation; Mice; Mice, Inbred C3H; Microfilariae; Proteins

Peripheral blood lymphocytes from patients with acute and chronic Wuchereria bancrofti infections responded poorly to concanavalin A, phytohaemagglutinin and pokeweed mitogen when cultured in heat-inactivated pooled normal serum. The lymphocyte response to mitogens in carriers of microfilariae (mff) were normal. The suppression of transformation to mitogens was not reversible by the removal of plastic adherent cells. Incubation with mitogens and the adult filarial worm antigen (BmA) did not alter the mitogen response either in control subjects or in filarial patients. The possible mechanism of immunosuppression is discussed.

Topics: Concanavalin A; Filariasis; Humans; In Vitro Techniques; Lymphocyte Activation; Phytohemagglutinins; Pokeweed Mitogens; T-Lymphocytes, Regulatory; Wuchereria bancrofti

Studies with inbred jirds chronically infected (greater than 5 months) with Brugia pahangi have demonstrated splenic suppressor cells which modulate in vitro responsiveness to mitogens and parasite antigens. The stimuli which induce suppression were characterized by analysing the effect of activated cells from inbred normal or B. pahangi-infected jirds on the PHA and PWM responsiveness of cultures on normal cells. Regulatory cells were stimulated in vitro with concanavalin A (Con A; 5 micrograms/ml) or an extract of adult B. pahangi (20 micrograms/ml) for 72 hr and irradiated (1500 rads) prior to cocultivation with normal cells. Addition of Con A-activated normal spleen cells to normal cells produced moderate suppression of PHA and enhancement of PWM responsiveness. However, Con A-stimulated spleen cells from infected animals consistently suppressed both the PHA and PWM responsiveness of normal cells by 80-90%. Spleen cells from chronically infected jirds were also induced by B. pahangi antigen to suppress both the PHA and PWM responsiveness of normal lymphocytes. In contrast, spleen cells from animals 3-15 weeks after infection and lymph node cells from all time points were capable of suppressing only PWM responses when stimulated by antigen. Normal spleen cells were not induced by B. pahangi antigens to exhibit immunoregulatory activity. The suppression mediated by antigen-induced spleen cells from chronically infected jirds was partially or totally alleviated by removal of non-specific suppressor cells which are plastic adherent and cyclophosphamide-sensitive, or by removal of antigen-specific suppressor cells which bear receptors for histamine. the results suggest the involvement of regulatory cell circuits in experimental filarial infections.

Topics: Animals; Antigens; Brugia; Concanavalin A; Filariasis; Filarioidea; Gerbillinae; Immune Tolerance; Lymphocyte Activation; Male; Spleen; T-Lymphocytes, Regulatory

Congenitally athymic nude (nu/nu) mice, immunologically reconstituted by thymus grafting before inoculation with infective larvae, and mice heterozygous for the nu gene (nu/+), mounted potent protective humoral and cellular immune responses to Brugia pahangi. Although responses were not identical, both groups of mice produced IgM, IgG and IgE antibodies specific for adult worm antigen (S-Ag) present in a crude aqueous extract, made immediate and delayed hypersensitivity footpad swelling responses when challenged with S-Ag and eliminated their infection in the early larval stages. Heterozygotes also exhibited a marked eosinophilia which peaked coincident with larval killing. In contrast, thymus grafting of patent nudes had no effect upon microfilaraemias or adult worm burdens and did not completely protect against a challenge larval inoculum although antibodies specific for S-Ag were produced. With the occasional exceptions of moderate immediate footpad swelling and very low titres of IgM specific for S-Ag, no specific immune responses to B. pahangi were found in ungrafted nude mice which allowed full development of adult worms and supported patent infections.

Topics: Animals; Antibody Formation; Brugia; Concanavalin A; Crosses, Genetic; Enzyme-Linked Immunosorbent Assay; Female; Filariasis; Heterozygote; Hypersensitivity; Immunoglobulin G; Immunoglobulin M; Leukocyte Count; Lymphocyte Activation; Lymphocytes; Male; Mice; Mice, Nude; Species Specificity

The in vitro immune responsiveness of lymphocytes from Brugia pahangi-infected jirds was examined after serial administration of cyclophosphamide (20 mg/kg). Cyclophosphamide had no effect on parasite burdens, anti-B. pahangi antibody titers, or suppressed spleen cell reactivity to B. pahangi antigens. Cyclophosphamide restored cellular responsiveness to the mitogens phytohemagglutinin, concanavalin A, and pokeweed mitogen.

Topics: Animals; Antibody Formation; Brugia; Concanavalin A; Cyclophosphamide; Filariasis; Gerbillinae; Lymphocytes; Male; Phytohemagglutinins; Pokeweed Mitogens

The dichotomy of resistance to Brugia pahangi (Nematoda: Filarioidea) between nonsusceptible, euthymic C3H/HeN mice, heterozygotic for the "nu" gene (+/nu), and susceptible, congenitally-athymic "nude" (nu/nu) C3H/HeN mice, suggests that resistance is thymus-dependent. To test this hypothesis, the effect of syngeneic neonatal thymus grafts and neonatal thymus cell suspensions on recovery of worms at day 40 PI, and responses to Concanavalin A (Con A) were examined in reconstituted nudes. Nude recipients of a thymus graft 7 or 14 wk before subcutaneous inoculation with 50 infective larvae (L3) yielded no worms and responded strongly to Con A. Serum from these mice reacted in two lines of identity with serum from similarly-infected heterozygotes by double radial immunodiffusion against an adult worm saline extract. Nude recipients of a thymus 2 days or 3 wk before inoculation harbored an average of three or two worms, respectively. Intravenous injection of nude recipients with 10(7) or 10(8) neonatal thymus cells seven weeks before inoculation was less effective in conferring resistance to B. pahangi and responsiveness to Con A. Complete resistance to B. pahangi could be adoptively transferred to nude mice by 10(8) spleen cells obtained from infection-primed heterozygotes and injected intravenously on the day of larval inoculation. The same numbers of worms were significantly reduced. less effective when injected 3 wk before inoculation, although numbers of worms were significantly reduced. Passive transfer of primed heterozygote serum, containing high titers of antibodies to adult worm and larval antigens, failed to protect nude recipients against a larval inoculum in the absence of cellular reconstitution.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Antibodies; Brugia; Concanavalin A; Female; Filariasis; Immunity, Innate; Immunization, Passive; Male; Mice; Mice, Inbred C3H; Mice, Nude; Spleen; T-Lymphocytes; Thymus Gland

Topics: Animals; Antigens; Binding Sites; Brugia; Chemical Fractionation; Concanavalin A; Enzyme-Linked Immunosorbent Assay; Epitopes; Female; Filariasis; Gerbillinae; Immunization; Immunization, Passive; Immunoglobulin G; Mice; Mice, Inbred Strains; Microfilariae; Solubility

Microfilariae of the rat filarial parasite Litomosoides carinii are killed by normal rat neutrophils which adhere to them in the presence of antibody and complement. Adherence and killing were increased in the presence of the fluid from 48-hr cultures of Con A- or PHA-stimulated normal rat lymph node cells. Both normal and augmented binding were inhibited by cytochalasin B. The factor(s) responsible for increased binding was non-dialysable, susceptible to heat (56 degrees C, 30 min) and freezing and thawing. Its production was inhibited by cycloheximide. Pretreatment of neutrophils with stimulated culture supernatant for 3 hr, followed by washing, also augmented adherence. Similar pretreatment also enhanced phagocytosis of opsonized sheep erythrocytes and latex particles but did not increase candidacidal activity. These experiments suggest that cell-mediated immune reactions leading to lymphokine production may potentiate anti-filarial antibody-dependent cellular cytotoxicity and general phagocytosis by neutrophils.

Topics: Animals; Antibodies; Cell Adhesion; Concanavalin A; Cycloheximide; Filariasis; Lymph Nodes; Lymphokines; Microfilariae; Neutrophils; Phagocytosis; Phytohemagglutinins; Rats

Topics: Animals; Antigens; Brugia; Concanavalin A; Filariasis; Gerbillinae; Immunosuppression Therapy; Lectins; Lymphocyte Activation; Male; Mitogens; Spleen