RESUMO
We and others have previously shown that nematodes or nematode products can stimulate or inhibit the generation of lymphocyte responses, suggesting that nematodes exert diverse effects on the developing immune responses of their host. In this study we examined the immunomodulatory effect of a soluble extract of Nippostrongylus brasiliensis (adult worm homogenate [AWH]) on B-cell responsiveness. We found that the extract inhibited the proliferation of B cells to lipopolysaccharide (LPS) stimulation in a dose-dependent manner. This effect was specific to B cells, since the extract did not inhibit T-cell proliferation to concanavalin A or anti-CD3 stimulation. The data presented here confirm that the extract is not toxic to B cells. We present evidence that the active factor is proteinaceous in nature and that the inhibitory activity is restricted to the adult stage of Nb. The extract does not appear to interfere with early activation events since it can be added up to 48 h after LPS stimulation, and it inhibited responses to phorbol myristate acetate and ionomycin. Furthermore, the proliferation of B cells to other activators was also inhibited by AWH. This observation shows that the inhibitory activity of AWH is not restricted to LPS-mediated B-cell proliferation. We present evidence that, in the absence of accessory cells, the inhibitory effect of the extract was ablated. This observation shows that the activity of AWH is not mediated directly on B cells but is mediated via the production of negative signals from accessory cells (macrophages), which affect a downstream pathway required by all B-cell activators tested. These effects on B-cell and accessory cell function are likely to have a significant effect on the outcome of infections experienced concurrently.
Assuntos
Linfócitos B/imunologia , Proteínas de Helminto/fisiologia , Ativação Linfocitária , Nippostrongylus/fisiologia , Animais , Feminino , Lipopolissacarídeos/farmacologia , Ativação de Macrófagos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteína Quinase C/fisiologia , Ratos , Ratos Sprague-DawleyRESUMO
Infection with the nematode parasite Nippostrongylus brasiliensis induces a pronounced type-2 T-cell response that is associated with marked polyclonal immunoglobulin E (IgE) and IgG1 production in mice. To examine the differential roles of the infection and products produced by nematodes, we investigated a soluble extract of N. brasiliensis for the ability to mediate this type-2 response. We found that the extract induced a marked increase in IgE and IgG1 levels, similar to that induced by the infection. The extract did not affect the level of IgG2a in serum, showing that the effect was specific to IgE and IgG1 (type-2-associated immunoglobulin) rather than inducing a nonspecific increase in all immunoglobulin isotypes. This response was also associated with increased interleukin-4 production in vitro. These results confirm that the extract, like infection, is a strong inducer of polyclonal type-2 responses and a reliable model for investigating the regulation of nematode-induced responses. The extract induced the production of IgG1 when added to in vitro cultures of lipopolysaccharide-stimulated B cells. This provides evidence for the induction of class switch. It did not induce upregulation of IgG1 in naive (unstimulated) B cells or expand B cells in in vitro cultures. Analysis of DNA from the spleens of mice treated with the extract by digestion-circularization PCR demonstrated a marked increase in the occurrence of gamma1 switch region gene recombination in the cells in vivo. These results provide strong evidence that soluble worm products are able to mediate the marked polyclonal gamma1/epsilon response and that infection is not required to mediate this response. Furthermore, these data provide evidence that the soluble nematode extract induces this effect by causing de novo class switch of B cells and not by an expansion of IgG1 B cells or an increase in antibody production by IgG1 plasma cells.
Assuntos
Switching de Imunoglobulina , Imunoglobulina E/biossíntese , Imunoglobulina G/biossíntese , Nippostrongylus/fisiologia , Animais , Feminino , Imunoglobulina G/classificação , Interleucina-13/biossíntese , Interleucina-4/biossíntese , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Five mouse strains, CBA/J, BALB/c, C3H/HeJ, A/J, and C57Bl/6J-bg-bg, all showed similar expulsion kinetics for Nippostrongylus brasiliensis (infective dose = 500 L3). Typically, parasite recovery was maximal on day 2 in the lungs and by day 4 in the small intestine. Few worms (less than 5% infective dose) were recovered on day 14 in all strains. These same mouse strains exhibited immune depression on day 5 of infection with mesenteric lymph node cells (MLN) showing reduced (10-30% normal) IgM, IgG, and IgA responses against heterologous antigen. The intestinal mast cell numbers and tissue histamine levels were examined in CBA/J mice. Mast cell numbers increased (normal = less than 1/villous crypt unit; VCU) from day 5 and peaked on day 12 (greater than 15/VCU). Intestinal histamine levels did not completely correlate with mast cell numbers with maximum concentrations (240 +/- 73 ng/g, 2-fold over normal) reached by day 8. Histamine concentrations in the intestine returned to normal levels by day 20.
Assuntos
Infecções por Nematoides/imunologia , Animais , Anticorpos Anti-Helmínticos/biossíntese , Contagem de Células , Feminino , Histamina/análise , Imunoglobulinas/biossíntese , Intestino Delgado/imunologia , Intestino Delgado/parasitologia , Intestino Delgado/patologia , Cinética , Pulmão/parasitologia , Masculino , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos , Infecções por Nematoides/parasitologia , Nippostrongylus/imunologia , Nippostrongylus/fisiologiaRESUMO
During acute inflammation, the mammalian liver responds with increased production and secretion of a series of plasma glycoproteins, collectively termed the acute-phase proteins, resulting from the release at the site of inflammation of polypeptide cytokines, including IL-1 and IL-6, which interact with receptors on hepatocytes and alter gene expression. This attribute of the systemic acute-phase response was studied throughout the course of infection with two nematode parasites in rats. Significant increases in serum haptoglobin, alpha 1-acid glycoprotein and alpha 1-cysteine protease inhibitor were detected coincident with episodes of skin, lung and intestinal pathology during Nippostrongylus brasiliensis, but were not seen during Trichinella spiralis, infection of the rat despite similar intestinal pathology. These changes were seen at both the protein and mRNA levels in the liver. Infection with T. spiralis was not anti-inflammatory, as macrophages from various sites could be induced in vitro to release inflammatory cytokines, and in vivo induction of inflammation by turpentine injection was similar in control and infected animals. However, macrophage populations recovered from animals infected with T. spiralis were not activated. Moreover, intestinal infection alone with intestinal stages of N. brasiliensis also failed to elicit the systemic acute-phase protein response, requiring an explanation involving skin and lung for the acute-phase response during gut inflammation in a primary infection with N. brasiliensis. Taken together, these data suggest that during the intestinal phase of nematode infection, with pathological changes to the gut, the systemic acute-phase response is not elicited through compromise or lack of stimulation of inflammatory cells in the intestine. The systemic parameters of the acute-phase response may not be a component of gastrointestinal pathology.
Assuntos
Reação de Fase Aguda/etiologia , Inflamação/etiologia , Infecções por Nematoides/imunologia , Proteínas de Fase Aguda/metabolismo , Proteínas de Fase Aguda/fisiologia , Animais , Enteropatias Parasitárias/imunologia , Fígado/análise , Masculino , Nippostrongylus , RNA Mensageiro/análise , Ratos , Ratos Endogâmicos , Triquinelose/imunologiaRESUMO
The cytochemical and functional characteristics of broncho-alveolar multinucleate giant cells and the kinetics of the giant cell response in the lungs of mice and rats during Nippostrongylus brasiliensis infection were studied. Primary infections resulted in significantly increased numbers of recoverable giant cells for up to 30 and 50 days in rats and mice, respectively. During secondary infections in the rat the giant cell response was more rapid and greater in magnitude than in a primary infection, suggesting that it was immunologically mediated. The giant cells displayed decreased C3- and IgG-dependent binding or phagocytic potential compared with mononucleate alveolar macrophages. Fusion of mononucleate alveolar macrophages into giant cells may therefore compromise complement and antibody dependent helminthocidal activity of these cells.