RESUMEN
The contribution of interleukin-3 (IL-3), a hematopoietic growth factor and immunoregulatory cytokine, to resistance to blood-stage malaria was investigated by infecting IL-3-deficient (knockout [KO]) mice with Plasmodium berghei NK65. Male IL-3 KO mice, but not female mice, were more resistant to infection than wild-type (WT) mice, as evidenced by lower peak parasitemia and prolonged survival. Both male and female IL-3 KO mice had increased splenomegaly and were more anemic than corresponding WT mice. Anemia was compensated for by an increase in bone marrow and splenic erythropoiesis in IL-3 KO mice, as evidenced by higher levels of erythroid progenitors. Plasma levels of gamma interferon (IFN-γ) and CXCL9 (monokine induced by IFN-γ [MIG]) were found to be significantly reduced in IL-3 KO mice during early stages of infection. In contrast, granulocyte colony-stimulating factor (G-CSF) levels were significantly higher, and the percentage of peripheral blood neutrophils lower, in infected IL-3 KO mice than in WT counterparts. Overall, our results indicate that IL-3 plays a critical role in suppressing protective immunity to P. berghei NK65 infection and that it is involved in inhibiting the development of splenomegaly, anemia, and erythropoiesis. IL-3 also influences IFN-γ, CXCL9, and G-CSF production in response to infection. The abnormal responses seen in infected IL-3 KO mice may be due to the lack of IL-3 during development, to the lack of IL-3 in the infected mature mice, or to both.
Asunto(s)
Interleucina-3/deficiencia , Interleucina-3/inmunología , Malaria/inmunología , Anemia/sangre , Anemia/inmunología , Anemia/metabolismo , Animales , Quimiocina CXCL9/sangre , Eritropoyesis/inmunología , Femenino , Factor Estimulante de Colonias de Granulocitos/sangre , Interferón gamma/sangre , Interleucina-3/metabolismo , Malaria/sangre , Malaria/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Neutrófilos/metabolismo , Plasmodium berghei/inmunología , Bazo/inmunología , Bazo/metabolismoRESUMEN
Interleukin-3 (IL-3) is an important hematopoietic growth factor and immunregulatory cytokine. Although activated T helper cells represent a main source of IL-3, other cell types have been reported to express this cytokine. However, precise identification and quantification of the cells that produce IL-3 in vivo have not been performed. Therefore, we used a CRISPR/Cas approach to engineer mice containing a bicistronic mRNA linking a readily identifiable reporter, enhanced green fluorescent protein (ZsGreen1), to IL-3 expression. To characterize these novel reporter mice, we first examined ZsGreen1 expression by CD4 T cells subsets primed and activated in vitro. We found that activated Th1 cells expressed â¼4-fold higher levels of ZsGreen1 as compared to Th0 and Th2 cells. Endogenous IL-3 expression remained intact although reporter Th1 cells secreted â¼33 % less IL-3 than similarly activated wild-type cells. To characterize the ability of reporter mice to accurately mark IL-3-producing cells in vivo, we infected mice with Nippostrongylus brasiliensis. Low but significant numbers of ZsGreen1+ CD4 T cells were detected in the mesenteric lymph nodes and lung following both primary and secondary infection. No difference in basophil and intestinal mast cell numbers were observed between infected reporter and wild-type mice indicating that reporter mice secreted IL-3 levels in vivo that results in IL-3-driven biological activities which are indistinguishable from those observed in corresponding wild-type mice. These IL-3 reporter mice will be a valuable resource to investigate IL-3-dependent immune responses in vivo.
Asunto(s)
Expresión Génica , Genes Reporteros , Interleucina-3/biosíntesis , Interleucina-3/genética , Ratones Transgénicos , Transgenes , Animales , Sistemas CRISPR-Cas , Femenino , Edición Génica , Orden Génico , Marcación de Gen , Vectores Genéticos/genética , Humanos , Activación de Linfocitos/genética , Activación de Linfocitos/inmunología , Masculino , Ratones , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismoRESUMEN
Enhanced basophil production is often associated with T(h)2-related conditions such as parasite infections or allergic inflammations. Our previous study demonstrated that T cell activation is necessary to promote basophil production in Nippostrongylus brasiliensis (Nb)-infected mice. Yet, mechanisms underlying how T cells aid infection-induced basophil production are not clear. In this report, we show that IL-3 produced by T cells activated by the infection enhances basophil production in Nb-infected mice. IL-3-deficient mice or Rag2-/- recipients of IL-3-deficient T cells but not of wild-type T cells failed to support basophil production following the Nb infection. Interestingly, although IL-3 was critical for preventing basophil apoptosis in vitro, IL-3 had little contribution to basophil survival and proliferation in vivo. Collectively, these results highlight a novel mechanism by which activation of adaptive immune components induces basophil production but not basophil survival via IL-3 production.
Asunto(s)
Basófilos/inmunología , Interleucina-3/metabolismo , Activación de Linfocitos/inmunología , Nippostrongylus/patogenicidad , Infecciones por Strongylida/inmunología , Linfocitos T/inmunología , Animales , Apoptosis , Basófilos/fisiología , Interleucina-3/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Infecciones por Strongylida/parasitología , Linfocitos T/metabolismoRESUMEN
Basophils represent potential effector and immunoregulatory cells, as well as a potential source of IL-4, during the immune response elicited by infection with the nematode Nippostrongylus brasiliensis (N.b.), and in other settings. However, the factors which regulate the numbers of blood basophils in mice, or the ability of these cells to produce IL-4, are not fully understood. We found that infection of mice with the nematodes N.b. or Strongyloides venezuelensis (S.v.) induced substantial increases in the numbers of blood basophils (to as high as 18% of circulating blood leukocytes). Experiments in IL-3-/- vs IL-3+/+ mice, and in IL-3-treated IL-3-/- mice, showed that essentially all of the increases in blood or bone marrow basophils during N.b. or S.v. infection were IL-3 dependent. Many of the blood, bone marrow or liver-derived basophils from IL-3-/- or IL-3+/+ mice expressed intracellular IL-4 upon stimulation with anti-IgE in vitro. However, after incubation of the cells with exogenous IgE in vitro, blood- or liver-derived basophils from IL-3+/+ mice exhibited higher levels of intracellular IL-4 after stimulation with anti-IgE than did basophils derived from IL-3-/- mice. Thus, IL-3 is a major regulator of the marked increases in blood basophil levels observed during infection of mice with N.b. or S.v. and also can enhance levels of intracellular IL-4 upon activation of basophils with anti-IgE in vitro.
Asunto(s)
Basófilos/inmunología , Inmunoglobulina E/inmunología , Interleucina-3/inmunología , Interleucina-4/inmunología , Animales , Ratones , Ratones Noqueados , Nippostrongylus , Receptores de IgE/inmunología , Infecciones por Strongylida/inmunología , Estrongiloidiasis/inmunologíaRESUMEN
Interleukin-3 (IL-3) and stem cell factor (SCF) are important mast cell growth and differentiation factors. Since both cytokines activate the transcription factor Stat5, a known regulator of proliferation and survival, we investigated the effects of Stat5 deficiency on mast cell development and survival. This article will review data presented at The Fourth International Workshop on Signal Transduction in the Activation and Development of Mast Cells and Basophils. The full set of data is now in preparation for publication. We find that the absence of Stat5 A and B results in a total loss of in vivo mast cell development. Bone marrow-derived mast cell (BMMC) populations can be cultured and maintained from Stat5-deficient mice in IL-3+SCF, but not in either cytokine alone. The absence of Stat5 resulted in aberrant control of Bcl-2, Bcl-x(L) and cyclin A2, with increased apoptosis and delayed cell cycle progression after IL-3 or SCF stimulation. These results indicate that Stat5 A and B are critical regulators of in vitro and in vivo mast cell biology.
Asunto(s)
Proteínas de Unión al ADN/fisiología , Mastocitos/inmunología , Proteínas de la Leche , Transactivadores/fisiología , Animales , Apoptosis , Degranulación de la Célula , Ciclinas/metabolismo , Citocinas/biosíntesis , Proteínas de Unión al ADN/genética , Interleucina-3/farmacología , Mastocitos/citología , Ratones , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Factor de Transcripción STAT5 , Factor de Células Madre/farmacología , Transactivadores/genéticaRESUMEN
Interleukin-3 (IL-3), which is derived from T cells and other sources, can promote the differentiation, proliferation, and migration of mast cells, basophils, and eosinophils. However, little is known about the ability of IL-3 to regulate the function of these cells in IgE-dependent and -independent allergic responses in vivo. Therefore, we sought to investigate the extent to which endogenously produced IL-3 can influence mast cell secretory function, the expression of local and systemic anaphylactic responses, and ragweed-induced eosinophilic peritonitis. We found that peritoneal mast cells from IL-3 deficient (IL-3 -/-) mice released less serotonin following challenge with low doses of anti-IgE antibody or antigen ex vivo than do cells isolated from corresponding wild-type (IL-3 +/+) mice. Both IL-3 -/- and +/+ mice expressed equivalent IgE-dependent passive cutaneous anaphylaxis responses following challenge with specific antigen and exhibited equivalent active systemic anaphylaxis responses to ovalbumin as assessed by changes in body temperature, death rates, total IgE production, and histamine release. In contrast, ragweed allergen immunization and peritoneal allergen challenge resulted in eosinophil recruitment that was greater in IL-3 -/- mice than in IL-3 +/+ mice. Our data demonstrates that IL-3 does not appear to be essential for local or systemic anaphylaxis. However, IL-3 production in vivo was found to enhance the mediator release from freshly isolated peritoneal mast cells stimulated ex vivo, and, unexpectedly, to inhibit the accumulation of eosinophils associated with a ragweed-induced allergic peritonitis model.
Asunto(s)
Anafilaxia/inmunología , Eosinófilos/inmunología , Interleucina-3/fisiología , Anafilaxis Cutánea Pasiva/inmunología , Peritonitis/inmunología , Rinitis Alérgica Estacional/inmunología , Alérgenos/inmunología , Ambrosia/inmunología , Anafilaxia/etiología , Animales , Modelos Animales de Enfermedad , Mastocitos/citología , Mastocitos/metabolismo , Ratones , Cavidad Peritoneal/citología , Serotonina/biosíntesisRESUMEN
Mast cells and MDSCs are increased by parasitic infection and tumor growth. We previously demonstrated that enhanced MDSC development in ADAM10 transgenic mice yielded resistance to Nb infection and that coculturing MDSCs and mast cells enhanced cytokine production. In the current work, we show that MDSC-mast cell coculture selectively enhances IgE-mediated cytokine secretion among mast cells, without increasing MDSC cytokine production. This effect was independent of cell contact and elicited by Ly6C(+) and Ly6C/G+ MDSC subsets. These interactions were functionally important. MDSC depletion with the FDA-approved drug gemcitabine exacerbated Nb or Trichinella spiralis infection and reduced mast cell-dependent AHR and lung inflammation. Adoptive transfer of MDSC worsened AHR in WT but not mast cell-deficient Wsh/Wsh mice. These data support the hypothesis that MDSCs enhance mast cell inflammatory responses and demonstrate that this interaction can be altered by an existing chemotherapeutic.
Asunto(s)
Inmunoglobulina E/inmunología , Mastocitos/inmunología , Células Mieloides/fisiología , Animales , Asma/inmunología , Células Cultivadas , Citocinas/biosíntesis , Femenino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Nippostrongylus/inmunología , Trichinella spiralis/inmunologíaRESUMEN
To evaluate the potential roles of Interleukin-3 (IL-3) and c-Kit, the tyrosine kinase receptor for stem cell factor (SCF), in eosinophil responses in vivo, we examined eosinophil numbers in uninfected or nematode-infected wild-type mice, IL-3-/- mice, and IL-3-/- mice that also have a marked reduction in SCF/c-Kit signaling (ie, Kit(W)/Kit(W-v), IL-3-/- mice). We found no significant differences in the numbers of eosinophils in the blood, bone marrow or various tissues of IL-3-/- vs IL-3+/+ mice, either at baseline or after the induction of bone marrow, blood or tissue eosinophilia in response to infection with Strongyloides venezuelensis (S.v.) or Nippostrongylus brasiliensis (N.b.). However, in mice with markedly impaired SCF/c-Kit signaling, IL-3 contributed significantly to the increased numbers of eosinophils that were observed in multiple tissues during S.v. infection, but not during infection with N.b.
Asunto(s)
Eosinofilia , Interleucina-3/inmunología , Proteínas Proto-Oncogénicas c-kit/inmunología , Estrongiloidiasis/inmunología , Animales , Basófilos/inmunología , Basófilos/parasitología , Modelos Animales de Enfermedad , Eosinofilia/inmunología , Eosinofilia/parasitología , Mastocitos/inmunología , Mastocitos/parasitología , Ratones , Ratones Noqueados , Nippostrongylus/inmunología , Transducción de Señal , Factor de Células Madre/inmunología , StrongyloidesRESUMEN
Interleukin-3 (IL-3) and stem cell factor (SCF) are important mast cell growth and differentiation factors. Since both cytokines activate the transcription factor signal transducer and activator of transcription 5 (Stat5), a known regulator of proliferation and survival, we investigated the effects of Stat5 deficiency on mast cell development and survival. Bone marrow-derived mast cell (BMMC) populations cultured from Stat5A/B-deficient mice survived in IL-3 + SCF, but not in either cytokine alone. These cells demonstrated reduced expression of Bcl-2, Bcl-x(L), cyclin A2, and cyclin B1, with increased apoptosis and delayed cell cycle progression during IL-3 or SCF culture. Finally, the absence of Stat5 resulted in loss of in vivo mast cell development, as judged by assessments of Stat5-deficient mice and transplantation of Stat5-deficient bone marrow cells to mast cell-deficient recipient mice. These results indicate that Stat5A and Stat5B are critical regulators of in vitro and in vivo mast cell development and survival.