RESUMEN
In contrast to mice from the majority of inbred strains, BALB mice develop aberrant Th2 responses and suffer progressive disease after infection with Leishmania major. These outcomes depend on the production of Interleukin 4, during the first 2 d of infection, by CD4+ T cells that express the Vbeta4-Valpha8 T cell receptors specific for a dominant I-A(d) restricted epitope of the LACK antigen from L. major. In contrast to this well established role of IL-4 in Th2 cell maturation, we have recently shown that, when limited to the initial period of activation of dendritic cells by L. major preceding T cell priming, IL-4 directs DCs to produce IL-12, promotes Th1 cell maturation and resistance to L. major in otherwise susceptible BALB/c mice. Thus, the antagonistic effects that IL-4 can have on Th cell development depend upon the nature of the cells (DCs or primed T cells) targeted for IL-4 signaling.
Asunto(s)
Interleucina-4/fisiología , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Ratones Endogámicos BALB C/inmunología , Animales , Antígenos de Protozoos/inmunología , Diferenciación Celular , Células Dendríticas/metabolismo , Predisposición Genética a la Enfermedad , Antígenos de Histocompatibilidad Clase II/inmunología , Interferón gamma/biosíntesis , Interferón gamma/genética , Interleucina-12/metabolismo , Interleucina-4/biosíntesis , Interleucina-4/genética , Ratones , Ratones Endogámicos BALB C/parasitología , Ratones Transgénicos , Modelos Animales , ARN Mensajero/biosíntesis , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Células TH1/citología , Células TH1/inmunología , Células Th2/citología , Células Th2/inmunologíaRESUMEN
Immunity to infection with intracellular pathogens is regulated by interleukin 12 (IL-12), which mediates protective T helper type 1 (TH1) responses, or IL-4, which induces TH2 cells and susceptibility. Paradoxically, we show here that when present during the initial activation of dendritic cells (DCs) by infectious agents, IL-4 instructed DCs to produce IL-12 and promote TH1 development. This TH1 response established resistance to Leishmania major in susceptible BALB/c mice. When present later, during the period of T cell priming, IL-4 induced TH2 differentiation and progressive leishmaniasis in resistant mice. Because immune responses developed via the consecutive activation of DCs and then T cells, the contrasting effects of IL-4 on DC development and T cell differentiation led to immune responses that had opposing functional phenotypes.
Asunto(s)
Células Dendríticas/efectos de los fármacos , Interleucina-4/fisiología , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Células TH1/inmunología , Animales , Antígenos de Protozoos/inmunología , Diferenciación Celular/efectos de los fármacos , Células Cultivadas/efectos de los fármacos , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Predisposición Genética a la Enfermedad , Inmunidad Innata , Interleucina-12/metabolismo , Interleucina-12/fisiología , Interleucina-4/farmacología , Leishmania major/fisiología , Ratones , Ratones Endogámicos BALB C , Ovalbúmina/inmunología , Proteínas Protozoarias/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Proteínas Recombinantes/farmacología , Organismos Libres de Patógenos Específicos , Células TH1/citología , Células Th2/citología , Células Th2/inmunología , Factores de TiempoRESUMEN
Early production of IL-4 by LACK-reactive Vbeta4-Valpha8 CD4(+) T cells instructs aberrant Th2 cell development and susceptibility to Leishmania major in BALB / c mice. This was demonstrated using Vbeta4(+)-deficient BALB / c mice as a result of chronic infection with MMTV (SIM), a mouse mammary tumor virus expressing a Vbeta4-specific superantigen. The early IL-4 response was absent in these mice which develop a Th1 response to L. major. Here, we studied the functional plasticity of LACK-reactive Vbeta4-Valpha8 CD4(+) T cells using BALB/ c mice inoculated with L. major shortly after infection with MMTV (SIM), i. e. before deletion of Vbeta4(+) cells. These mice fail to produce the early IL-4 response to L. major and instead exhibit an IFN-gamma response that occurs within LACK-reactive Vbeta4-Valpha8 CD4(+) T cells. Neutralization of IFN-gamma restores the production of IL-4 by these cells. These data suggest that the functional properties of LACK-reactive Vbeta4-Valpha8 CD4(+) T cells are not irreversibly fixed.
Asunto(s)
Antígenos de Protozoos , Regiones Determinantes de Complementariedad/inmunología , Interleucina-4/inmunología , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Proteínas Protozoarias/inmunología , Células Th2/inmunología , Animales , Anticuerpos/inmunología , Anticuerpos/farmacología , Diferenciación Celular/efectos de los fármacos , Femenino , Interferón gamma/antagonistas & inhibidores , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-4/biosíntesis , Interleucina-4/genética , Leishmania major/crecimiento & desarrollo , Leishmania major/fisiología , Leishmaniasis Cutánea/parasitología , Activación de Linfocitos/efectos de los fármacos , Recuento de Linfocitos , Virus del Tumor Mamario del Ratón/genética , Ratones , Ratones Endogámicos BALB C , ARN Mensajero/genética , ARN Mensajero/metabolismo , Superantígenos/genética , Superantígenos/inmunología , Células Th2/citología , Células Th2/efectos de los fármacos , Células Th2/metabolismo , Factores de Tiempo , Transducción Genética , Regulación hacia ArribaRESUMEN
The possible immunomodulatory role of polymorphonuclear leukocytes (PMN) in CD4+ T lymphocyte differentiation in mice was examined by studying the effect of transient depletion of PMN during the early phase after Leishmania major delivery. A single injection of the PMN-depleting NIMP-R14 mAb 6 h before infection with L. major prevented the early burst of IL-4 mRNA transcription otherwise occurring in the draining lymph node of susceptible BALB/c mice. Since this early burst of IL-4 mRNA transcripts had previously been shown to instruct Th2 differentiation in mice from this strain, we examined the effect of PMN depletion on Th subset differentiation at later time points after infection. The transient depletion of PMN in BALB/c mice was sufficient to inhibit Th2 cell development otherwise occurring after L. major infection. Decreased Th2 responses were paralleled with partial resolution of the footpad lesions induced by L. major. Furthermore, draining lymph node-derived CD4+ T cells from PMN-depleted mice remained responsive to IL-12 after L. major infection, unlike those of infected BALB/c mice receiving control Ab. PMN depletion had no effect when the NIMP-R14 mAb was injected 24 h postinfection. The protective effect of PMN depletion was shown to be IL-12 dependent, as concomitant neutralization of IL-12 reversed the protective effect of PMN depletion. These results suggest a role for an early wave of PMN in the development of the Th2 response characteristic of mice susceptible to infection with L. major.
Asunto(s)
Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Neutrófilos/inmunología , Células Th2/metabolismo , Animales , Anticuerpos Monoclonales/administración & dosificación , Diferenciación Celular/inmunología , Movimiento Celular/inmunología , Citocinas/biosíntesis , Citocinas/genética , Susceptibilidad a Enfermedades , Inmunidad Celular , Inmunidad Innata , Inyecciones Intraperitoneales , Interferón gamma/antagonistas & inhibidores , Interferón gamma/inmunología , Interleucina-12/antagonistas & inhibidores , Interleucina-12/inmunología , Interleucina-12/fisiología , Leishmaniasis Cutánea/patología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/metabolismo , Ganglios Linfáticos/patología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Neutropenia/inmunología , Neutropenia/patología , Neutrófilos/parasitología , Neutrófilos/patología , ARN Mensajero/biosíntesis , Células Th2/inmunología , Células Th2/parasitología , Células Th2/patología , Factores de TiempoRESUMEN
In contrast to intact BALB/c mice, BALB/c mice rendered deficient in Vbeta4+ CD4+ T cells develop a Th1 response to infection with Leishmania major and are resistant. Vbeta4-deficient BALB/c mice are unable to generate the early IL-4 transcription occurring in Vbeta4 Valpha8 CD4+ T cells of BALB/c mice within 1 day of infection. Here we demonstrate that treatment of Vbeta4-deficient BALB/c mice with IL-4 during the first 64 h after infection instructs Th2 cell development and susceptibility to infection. The demonstrated inability of IL-4 to reverse the resistant phenotype of BALB/c mice treated with anti-CD4 mAb the day before infection suggest that these effects of IL-4 require its interaction with CD4+ T cells. In contrast to draining lymph node cells from BALB/c mice, cells from Vbeta4-deficient BALB/c mice remain responsive to IL-12 following infection. Strikingly, administration of IL-4 to Vbeta4-deficient BALB/c mice renders their lymph node cells unresponsive to IL-12 by down-regulating IL-12R beta2-chain expression. This study directly demonstrates that in BALB/c mice IL-4 is necessary and sufficient to initiate the molecular events steering Th2 cell maturation and susceptibility to L. major.
Asunto(s)
Interleucina-4/biosíntesis , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Células Th2/metabolismo , Animales , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD4-Positivos/patología , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Progresión de la Enfermedad , Regulación hacia Abajo/inmunología , Femenino , Tolerancia Inmunológica/genética , Inmunidad Innata/genética , Inyecciones Intraperitoneales , Interleucina-12/metabolismo , Interleucina-12/farmacología , Interleucina-4/administración & dosificación , Leishmaniasis Cutánea/genética , Ganglios Linfáticos/citología , Ganglios Linfáticos/inmunología , Linfopenia/genética , Linfopenia/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Receptores de Interleucina/antagonistas & inhibidores , Receptores de Interleucina/biosíntesis , Receptores de Interleucina-12 , Células Th2/inmunologíaRESUMEN
The first experimental evidence for the development of polarized CD4+ Th1 and Th2 responses in vivo has been obtained using the murine model of infection with Leishmania major, an intracellular parasite of macrophages in their vertebrate host. Genetically determined resistance and susceptibility to infection with this parasite have been clearly demonstrated to result from the development of polarized Th1 and Th2 responses, respectively. Using this model system, the dominant role of cytokines in the induction of polarized CD4+ responses has been validated in vivo. The requisite role of IL-4 in mediating both Th2 differentiation and susceptibility to infection in BALB/c mice has directed interest towards the search for evidence of IL-4 production early after infection and identification of its cellular source. We have been able to demonstrate a burst of IL-4 production in susceptible BALB/c mice within the first day of infection with L. major and could establish that this rapidly produced IL-4 instructed Th2 lineage commitment of subsequently activated CD4+ T cells and stabilized this commitment by downregulating IL-12 Rbeta2 chain expression, resulting in susceptibility to infection. Strikingly, this early IL-4 response to infection resulted from the cognate recognition of a single epitope in a distinctive antigen, LACK, from this complex microorganism by a restricted population of CD4+ T cells that express Vbeta4-Valpha8 T cell receptors.
Asunto(s)
Leishmania major/patogenicidad , Leishmaniasis/inmunología , Células Th2/citología , Animales , Citocinas/biosíntesis , Citocinas/inmunología , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Células Th2/parasitologíaRESUMEN
Experimental leishmaniasis offers a well characterized model of T helper type 1 cell (Th1)-mediated control of infection by an intracellular organism. Susceptible BALB/c mice aberrantly develop Th2 cells in response to infection and are unable to control parasite dissemination. The early CD4(+) T cell response in these mice is oligoclonal and reflects the expansion of Vbeta4/ Valpha8-bearing T cells in response to a single epitope from the parasite Leishmania homologue of mammalian RACK1 (LACK) antigen. Interleukin 4 (IL-4) generated by these cells is believed to direct the subsequent Th2 response. We used T cells from T cell receptor-transgenic mice expressing such a Vbeta4/Valpha8 receptor to characterize altered peptide ligands with similar affinity for I-Ad. Such altered ligands failed to activate IL-4 production from transgenic LACK-specific T cells or following injection into BALB/c mice. Pretreatment of susceptible mice with altered peptide ligands substantially altered the course of subsequent infection. The ability to confer a healer phenotype on otherwise susceptible mice using altered peptides that differed by a single amino acid suggests limited diversity in the endogenous T cell repertoire recognizing this antigen.
Asunto(s)
Antígenos de Protozoos/inmunología , Epítopos/inmunología , Antígenos de Histocompatibilidad Clase II/inmunología , Leishmania major/inmunología , Fragmentos de Péptidos/inmunología , Proteínas Protozoarias/inmunología , Células Th2/inmunología , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Animales , Susceptibilidad a Enfermedades , Femenino , Tolerancia Inmunológica , Inmunidad Celular , Interferón gamma/metabolismo , Interleucina-4/metabolismo , Leishmaniasis Cutánea/inmunología , Ligandos , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Datos de Secuencia Molecular , Fragmentos de Péptidos/síntesis química , Proteínas Protozoarias/química , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Proteínas Recombinantes de Fusión/inmunología , Superantígenos/inmunologíaAsunto(s)
Linfocitos T CD4-Positivos/inmunología , Susceptibilidad a Enfermedades/inmunología , Interleucina-12/inmunología , Interleucina-4/inmunología , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Células Th2/inmunología , Animales , Antígenos de Protozoos/inmunología , Linfocitos T CD4-Positivos/citología , Diferenciación Celular , Interleucina-4/biosíntesis , Interleucina-4/genética , Ratones , Ratones Endogámicos BALB C , Proteínas Protozoarias/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/inmunologíaRESUMEN
Within 1 day of infection with Leishmania major, susceptible BALB/c mice produce a burst of IL-4 in their draining lymph nodes, resulting in a state of unresponsiveness to IL-12 in parasite-specific CD4+ T cells within 48 h. In this report we examined the molecular mechanism underlying this IL-12 unresponsiveness. Extinction of IL-12 signaling in BALB/c mice is due to a rapid down-regulation of IL-12R beta2-chain mRNA expression in CD4+ T cells. In contrast, IL-12R beta2-chain mRNA expression was maintained on CD4+ T cells from resistant C57BL/6 mice. The down-regulation of the IL-12R beta2-chain mRNA expression in BALB/c CD4+ T cells is a consequence of the early IL-4 production. In this murine model of infection, a strict correlation is shown in vivo between expression of the IL-12R beta2-chain in CD4+ T cells and the development of a Th1 response and down-regulation of the mRNA beta2-chain expression and the maturation of a Th2 response. Treatment of BALB/c mice with IFN-gamma, even when IL-4 has been produced for 48 h, resulted in maintenance of IL-12R beta2-chain mRNA expression and IL-12 responsiveness. The data presented here support the hypothesis that the genetically determined susceptibility of BALB/c mice to infection with L. major is primarily based on an up-regulation of IL-4 production, which secondarily induces extinction of IL-12 signaling.
Asunto(s)
Linfocitos T CD4-Positivos/metabolismo , Regulación hacia Abajo/inmunología , Tolerancia Inmunológica , Interleucina-4/biosíntesis , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Receptores de Interleucina/biosíntesis , Animales , Femenino , Inyecciones Intraperitoneales , Interferón gamma/administración & dosificación , Interferón gamma/fisiología , Interleucina-4/fisiología , Leishmaniasis Cutánea/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos , Ratones Noqueados , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/biosíntesis , Receptores de Interleucina/administración & dosificación , Receptores de Interleucina/genética , Receptores de Interleucina/inmunología , Receptores de Interleucina-12 , Células Th2/metabolismo , Transcripción Genética/inmunologíaRESUMEN
BACKGROUND & AIMS: Infection with Helicobacter induces a T helper type 1 response in mice and humans. Mice can be cured or protected from infection with Helicobacter by mucosal immunization with recombinant H. pylori urease B subunit (rUreB). This study characterizes the immune response of infected mice immunized with rUreB. METHODS: BALB/c mice were infected with H. felis. Two weeks later, they were orally immunized four times with rUreB and cholera toxin (CT) at weekly intervals. Controls were only infected or sham-immunized with CT. Animals were killed at various times after immunization. Splenic CD4(+) cells were obtained and cultured in vitro with rUreB to evaluate antigen-specific proliferation and induction of interferon gamma and interleukin 4 secretion. RESULTS: All rUreB-immunized mice (n = 8) were cured from infection 3 weeks after the fourth immunization. Immunization induced a proliferative response of splenic CD4(+) cells, a progressive decrease in interferon gamma secretion, and a concomitant increase in interleukin 4 secretion after each immunization. A simultaneous increase in rUreB specific serum immunoglobulin G1 levels was observed in infected/immunized mice. CONCLUSIONS: In BALB/c mice, therapeutic mucosal immunization with rUreB induces progressively a Th2 CD4(+) T cell response resulting in the elimination of the pathogen.
Asunto(s)
Infecciones por Helicobacter/inmunología , Helicobacter/enzimología , Inmunización , Isoenzimas/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Ureasa/inmunología , Animales , Formación de Anticuerpos/inmunología , División Celular/efectos de los fármacos , Toxina del Cólera/inmunología , Citocinas/metabolismo , Femenino , Mucosa Gástrica/inmunología , Mucosa Gástrica/metabolismo , Mucosa Gástrica/patología , Infecciones por Helicobacter/terapia , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes , Linfocitos T/patología , Células TH1/patología , Células Th2/patologíaRESUMEN
The murine model of infection with Leishmania major has allowed the demonstration of a causal relationship between, on the one hand, genetically determined resistance to infection and the development of a Th1 CD4+ cell response, and on the other hand, genetically determined susceptibility and Th2 cell maturation. Using this murine model of infection, the role of cytokines in directing the functional differentiation pathway of CD4+ T cell precursors, has been demonstrated in vivo. Thus, IL-12 and IFN-gamma have been shown to favour Th1 cell development and IL-4 is crucial for the differentiation of Th2 responses. Maturation of a Th2 response in susceptible BALB/c mice following infection with L. major is triggered by the IL-4 produced during the first two days after parasite inoculation. This IL-4 rapidly renders parasite specific CD4+ T cells precursors unresponsive to IL-12. A restricted population of CD4+ T cells expressing the V beta 4V alpha 8 TCR heterodimer and recognizing a single epitope on the LACK (Leishmania Activated C-Kinase) antigen of L. major is responsible for this rapid production of IL-4, instructing subsequent differentiation towards the Th2 phenotype of CD4+ T cells specific for several parasite antigens.
Asunto(s)
Leishmania major/inmunología , Leishmaniasis Cutánea/genética , Leishmaniasis Cutánea/inmunología , Animales , Linfocitos T CD4-Positivos/inmunología , Diferenciación Celular , Predisposición Genética a la Enfermedad , Humanos , Inmunidad Innata/genética , Interleucina-4/biosíntesis , Líquido Intracelular , Ratones , Ratones Endogámicos BALB C , Células TH1/inmunología , Células Th2/citología , Células Th2/inmunologíaRESUMEN
Mouse mammary tumor virus (MMTV) is a retrovirus which induces a strong immune response and a dramatic increase in the number of infected cells through the expression of a superantigen (SAg). Many cytokines are likely to be involved in the interaction between MMTV and the immune system. In particular, alpha/beta interferon (IFN-alpha/beta) and gamma interferon (IFN-gamma) exert many antiviral and immunomodulatory activities and play a critical role in other viral infections. In this study, we have investigated the importance of interferons during MMTV infection by using mice with a disrupted IFN-alpha/beta or IFN-gamma receptor gene. We found that the SAg response to MMTV was not modified in IFN-alpha/betaR(0/0) and IFN-gammaR(0/0) mice. This was true both for the early expansion of B and T cells induced by the SAg and for the deletion of SAg-reactive cells at later stages of the infection. In addition, no increase in the amount of proviral DNA was detected in tissues of IFN-alpha/betaR(0/0) and IFN-gammaR(0/0) mice, suggesting that interferons are not essential antiviral defense mechanisms during MMTV infection. In contrast, IFN-gammaR(0/0) mice had increased amounts of IL-4 mRNA and an altered usage of immunoglobulin isotypes with a reduced frequency of IgG2a- and IgG3-producing cells. This was associated with lower titers of virus-specific antibodies in serum early after infection, although efficient titers were reached later.
Asunto(s)
Virus del Tumor Mamario del Ratón/inmunología , Receptores de Interferón/inmunología , Infecciones por Retroviridae/inmunología , Infecciones Tumorales por Virus/inmunología , Animales , Anticuerpos Antivirales/inmunología , Antígenos Virales/inmunología , Cruzamientos Genéticos , ADN Viral , Femenino , Eliminación de Gen , Expresión Génica , Interferón gamma/biosíntesis , Interferón gamma/genética , Interleucina-4/biosíntesis , Interleucina-4/genética , Masculino , Proteínas de la Membrana , Ratones , Ratones Endogámicos BALB C , Reacción en Cadena de la Polimerasa , Provirus/genética , Receptor de Interferón alfa y beta , Receptores de Interferón/genética , Superantígenos/inmunología , Factores de Tiempo , Receptor de Interferón gammaRESUMEN
The essential role of cytokines in parasitic diseases has been emphasised since the in vivo description of the importance of T helper 1 (Th1) and T helper 2 (Th2) CD4+ T cell responses in resistance and susceptibility to infection with L. major in mice. Th1 cells produced IL-2, IFN-gamma and Lymphotoxin T (LT) and Th2 cells produce IL-4, IL-5 and IL-13. In this model of infection the correlation between on the one hand resistance to infection and the development of a Th1 response and on the other hand susceptibility and Th2 cell development allowed the identification of the mechanisms directing the differentiation of CD4+ T cell precursors towards either Th1 type or Th2 type responses. Cytokines are the crucial inducer of functional CD4+ T cell subset differentiation during infection with L. major. IL-12 and IFN-gamma direct the differentiation of Th1 response and IL-4 of a Th2 response. In susceptible mice, careful analysis of IL-4 production during the first days of infection has shown that the IL-4 produced as a result of a very early burst of IL-4 mRNA expression (16 hours) plays a essential role in the maturation of a Th2 CD4+ T cell response by rendering the CD4+ T cell precursors unresponsive to IL-12. Activation of a restricted population of CD4+ T cells expressing the V beta 4 V alpha 8 TCR heterodimer after recognition of a single antigen, the LACK (Leishmania Activated c Kinase) antigen, resulted in this rapid production of IL-4 required for the subsequent CD4+ T cell differentiation. Thus, tolerization of these cells might contribute a strategy for preventing infection with L. major.
Asunto(s)
Citocinas/inmunología , Leishmaniasis Cutánea/inmunología , Animales , Linfocitos T CD4-Positivos/inmunología , Modelos Animales de Enfermedad , Leishmania major/inmunología , RatonesRESUMEN
Resistance and susceptibility to infection with the intracellular parasite, Leishmania major, are mediated by parasite-specific CD4+ Th1 and Th2 cells, respectively. It is well established that the protective effect of parasite-specific CD4+ Th1 cells is largely dependent upon the IFN-gamma produced. However, recent results indicate that the effect of Th1 cells on resolution of lesions induced by L. major in genetically resistant mice also requires a functional Fas-FasL pathway of cytotoxicity. In contrast to resistant mice, susceptible BALB/c mice develop aberrant Th2 responses following infection with L. major and consequently suffer progressive disease. These outcomes clearly depends upon the production of interleukin 4 (IL-4) early after infection. We have shown that a burst of IL-4 mRNA, peaking in draining lymph nodes of BALB/c mice 16 hrs after infection, occurs within CD4+ T cells that express V beta 4-V alpha 8 T cell receptors. In contrast to control and V beta 6-deficient mice, V beta 4-deficient BALB/c mice were resistant to infection, demonstrating the role of these cells in Th2 development. The early IL-4 response was absent in these mice, and Th1 responses occurred following infection. The LACK antigen of L. major induced comparable IL-4 production in V beta 4-V alpha 8 CD4+ T cells. Thus, the IL-4 required for Th2 development and susceptibility to L. major is produced by a restricted population of V beta 4-V alpha 8 CD4+ T cells after cognate interaction with a single antigen from this complex parasite. The IL-4 produced rapidly by these CD4+ T cells induces within 48 hours a state of unresponsiveness to IL-12 among parasite-specific CD4+ T cell precursors by downregulating the IL-12 receptor beta 2 chain expression.
Asunto(s)
Citotoxicidad Inmunológica , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Células TH1/inmunología , Células Th2/inmunología , Animales , Antígenos CD4/inmunología , Susceptibilidad a Enfermedades/inmunología , Ratones , Ratones Endogámicos BALB CRESUMEN
Soluble Leishmania antigen (SLA) from both developmental stages of L. major (L. major MRHO/IR/75/ER) were prepared. Three and five subfractions of SLA from amastigote and promastigote were obtained by fast protein liquid chromatography (FPLC), respectively. Biochemical analyses and comparison of amastigote and promastigote SLA were done. The biochemical analyses revealed that the first fraction of L. major amastigote possesses a distinct band on its electrophoretic mobility pattern corresponding to a position of 24 kD, and it has enzymatic activity with characteristics of a cysteine proteinase. The isolated fractions of amastigote were tested for induction of proliferation, interferon-gamma (IFN-gamma) and IL-4 production in cultures of peripheral blood mononuclear cells (PBMC) from individuals who had recovered and also chronic patients of cutaneous leishmaniasis caused by L. major. The cells of recovered individuals compared with chronic cases proliferated profoundly in response to the first fraction of amastigote SLA. In all recovered individuals, the IFN-gamma, but not IL-4, was secreted in response to stimulation with the first fraction of amastigote SLA. In chronic cutaneous leishmaniasis, IFN-gamma was infrequently observed in response to stimulation by all three fractions of amastigote SLA, but secretion of IL-4 was observed. These data indicate that first fraction of amastigote SLA is a strong inducer of primed human immune response to L. major, and may have a protective function.
Asunto(s)
Antígenos de Protozoos/inmunología , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Animales , Antígenos de Protozoos/análisis , Femenino , Humanos , Interferón gamma/inmunología , Interleucina-4/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones DesnudosRESUMEN
BALB/c mice develop aberrant T helper 2 (Th2) responses and suffer progressive disease after infection with Leishmania major. These outcomes depend on the production of interleukin-4 (IL-4) early after infection. Here we demonstrate that the burst of IL-4 mRNA, peaking in draining lymph nodes of BALB/c mice 16 hr after infection, occurs within CD4+ T cells that express V beta 4 V alpha 8 T cell receptors. In contrast to control and V beta 6-deficient BALB/c mice, V beta 4-deficient BALB/c mice were resistant to infection, demonstrating the role of these cells in Th2 development. The early IL-4 response was absent in these mice, and T helper 1 responses occurred following infection. Recombinant LACK antigen from L. major induced comparable IL-4 production in V beta 4 V alpha 8 CD4+ cells. Thus, the IL-4 required for Th2 development and susceptibility to L. major is produced by a restricted population of V beta 4 V alpha 8 CD4+ T cells after cognate interaction with a single antigen from this complex organism.
Asunto(s)
Antígenos CD4/análisis , Interleucina-4/biosíntesis , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Receptores de Antígenos de Linfocitos T alfa-beta/inmunología , Subgrupos de Linfocitos T/metabolismo , Células Th2/citología , Animales , Antígenos de Protozoos/farmacología , Antígenos CD4/genética , Diferenciación Celular/inmunología , Susceptibilidad a Enfermedades , Femenino , Inmunidad Innata , Interleucina-4/genética , Ratones , Ratones Endogámicos BALB C , Proteínas Protozoarias/farmacología , ARN Mensajero/biosíntesis , Receptores de Antígenos de Linfocitos T alfa-beta/metabolismo , Proteínas Recombinantes/farmacología , Subgrupos de Linfocitos T/inmunologíaRESUMEN
An unusual subset of mature T cells expresses natural killer (NK) cell-related surface markers such as interleukin-2 receptor beta (IL-2R beta; CD122) and the polymorphic antigen NK1.1. These "NK-like" T cells are distinguished by their highly skewed V alpha and V beta repertoire and by their ability to rapidly produce large amounts of IL-4 upon T cell receptor (TCR) engagement. The inbred mouse strain SJL (which expresses NK1.1 on its NK cells) has recently been reported to lack NK1.1+ T cells and consequently to be deficient in IL-4 production upon TCR stimulation. We show here, however, that SJL mice have normal numbers of IL-2R beta+ T cells with a skewed V beta repertoire characteristic of "NK-like" T cells. Furthermore lack of NK1.1 expression on IL-2R beta+ T cells in SJL mice was found by backcross analysis to be controlled by a single recessive gene closely linked to the NKR-P1 complex on chromosome 6 (which encodes the NK1.1 antigen). Analysis of a panel of inbred mouse strains further demonstrated that lack of NK1.1 expression on IL-2R beta+ T cells segregated with NKR-P1 genotype (as assessed by restriction fragment length polymorphism) and thus was not restricted to the SJL strain. In contrast, defective TCR induced IL-4 production (which appeared to be a unique property of SJL mice) seems to be controlled by two recessive genes unlinked to NKR-P1. Collectively, our data indicate that "NK-like" T cells develop normally in SJL mice despite genetically distinct defects in NK1.1 expression and inducible IL-4 production.
Asunto(s)
Antígenos/genética , Interleucina-4/genética , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Lectinas Tipo C , Proteínas/genética , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo , Animales , Antígenos/biosíntesis , Antígenos Ly , Antígenos de Superficie/genética , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Ligamiento Genético/inmunología , Genotipo , Interleucina-4/biosíntesis , Hígado/inmunología , Hígado/patología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Ratones Endogámicos DBA , Ratones Endogámicos NZB , Subfamilia B de Receptores Similares a Lectina de Células NK , Biosíntesis de ProteínasRESUMEN
Previous results have documented a burst of IL-4 mRNA that peaks in draining lymph nodes of susceptible BALB/c mice 16 h after infection with Leishmania major. The importance of this early IL-4 response in subsequent Th2 cell maturation is supported by observations showing that 1) neutralization of IL-4 at the initiation of infection or 2) administration of IL-12, which results in an inhibition of the 16 h IL-4 mRNA burst, inhibits Th2 cell development. However, both treatments are effective in hampering Th2 cell development only if given at a time when IL-4 has been produced for <48 h. At this time after infection, lymph node CD4+ T cells from BALB/c mice no longer respond to IL-12. This IL-12 unresponsiveness is prevented in mice treated with anti-IL-4 Abs at the initiation of infection. Finally, the inhibition of Th2 development in BALB/c mice treated with anti-IL-4 Abs at the onset of infection results from maintenance of IL-12 responsiveness, since it requires IL-12. Together, these results reveal a narrow window of time, between 16 h and <48 h after infection, during which IL-4 produced rapidly in BALB/c mice renders T cells unresponsive to IL-12, allowing their differentiation toward the Th2 phenotype.
Asunto(s)
Interleucina-12/biosíntesis , Interleucina-4/biosíntesis , Leishmania major/inmunología , Leishmaniasis Cutánea/etiología , Leishmaniasis Cutánea/inmunología , Animales , Anticuerpos Monoclonales/farmacología , Susceptibilidad a Enfermedades , Femenino , Inmunidad Innata , Interleucina-12/farmacología , Interleucina-4/genética , Interleucina-4/inmunología , Cinética , Leishmaniasis Cutánea/genética , Ganglios Linfáticos/citología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , ARN Mensajero/biosíntesisRESUMEN
Leishmania spp. are intracellular protozoan parasites that are delivered within the dermis of their vertebrate hosts. Within this peripheral tissue and the draining lymph node, they find and/or rapidly create dynamic microenvironments that determine their ultimate fate, namely their more or less successful expansion, and favour their transmission to another vertebrate host though a blood-feeding vector. Depending on their genetic characteristics as well as the genetic make-up of their hosts, once within the dermis Leishmania spp. very rapidly drive and maintain sustained T cell-dependent immune responses that arbitrate their ultimate fate within their hosts. The analysis of the parasitism exerted by Leishmania major in mice of different genetic backgrounds has allowed us to recognize some of the early and late mechanisms driven by this parasite that lead to either uncontrolled or restricted parasitism. Uncontrolled parasitism by Leishmania major characterizing mice from a few inbred strains (e.g. BALB/c) is associated with the expansion of parasite reactive Th2 CD4 lymphocytes and results from their rapid and sustained activity. In contrast, restricted parasitism characteristic of mice from the majority of inbred strains results from the development of a polarized parasite-specific Th1 CD4 response. This murine model of infection has already been and will continue to be particularly instrumental in dissecting the rules controlling the pathway of differentiation of T cells in vivo. In the long run, the understanding of these rules should contribute to the rational development of novel immunotherapeutic interventions against severe infectious diseases.
Asunto(s)
Citocinas/biosíntesis , Leishmania major/inmunología , Leishmaniasis Cutánea/inmunología , Leishmaniasis Cutánea/parasitología , Linfocitos T/inmunología , Animales , Modelos Animales de Enfermedad , Predisposición Genética a la Enfermedad , Interacciones Huésped-Parásitos , Leishmania major/patogenicidad , Ratones , Ratones Endogámicos , VirulenciaRESUMEN
Mice with homologous disruption of the gene coding for either the p35 subunit or the p40 subunit of interleukin-12 (IL-12) and derived from a strain genetically resistant to infection with Leishmania major have been used to study further the role of this cytokine in resistance to infection and the differentiation of functional CD4+ T cell subsets in vivo. Wild-type 129/Sv/Ev mice are resistant to infection with L. major showing only small lesions which resolve spontaneously within a few weeks and develop a type 1 CD4+ T cell response. In contrast, mice lacking bioactive IL-12 (IL-12p35-/- and IL-12p40-/-) developed large, progressing lesions. Whereas resistant mice were able to mount a delayed-type hypersensitivity (DTH) response to Leishmania antigen, susceptible BALB/c mice as well as IL-12-deficient 129/Sv/Ev mice did not show any DTH reaction. To characterize the functional phenotype of CD4+ T cells triggered in infected wild-type mice and IL-12-deficient mice, the expression of mRNA for interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) in purified CD4+ lymph node cells was analyzed. Wild-type 129/Sv/Ev mice showed high levels of mRNA for IFN-gamma and low levels of mRNA for IL-4 which is indicative of a Th1 response. In contrast, IL-12- deficient mice and susceptible BALB/c mice developed a strong Th2 response with high levels of IL-4 mRNA and low levels of IFN-gamma mRNA in CD4+ T cells. Similarly, lymph node cells from infected wild-type 129 mice produced predominantly IFN-gamma in response to stimulation with Leishmania antigen in vitro whereas lymph node cells from IL-12-deficient mice and susceptible BALB/c mice produced preferentially IL-4. Taken together, these results confirm in vivo the importance of IL-12 in induction of Th1 responses and protective immunity against L. major.