RESUMEN
As an immune privilege site, there are various types of immune cells in the testis. Previous research has been focused on the testicular macrophages, and much less is known about the T cells in the testis. Here, we found that T cells with memory phenotypes were the most abundant leukocyte in the testis except for macrophages. Our results showed that the proportion of testicular T cells increases gradually from birth to adulthood in mice and that the primary type of T cells changed from γδTCR+ T cells to αßTCR+ T cells. In addition, under homeostatic conditions, CD8+ T cells are the dominant subgroup and have different phenotypic characteristics from CD4+ T cells. We found that cDC1, but not cDC2, is necessary for the presence of T cells in the testis under physiological state. A significant decrease of T cells does not have a deleterious effect on the development of the testis or spermatogenesis. However, cDC1-dependent T cells play an indispensable role in chronic autoimmune orchitis of the testis. Collectively, our multifaceted data provide a comprehensive picture of the accumulation, localization, and function of testicular T cells.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Linfocitos T CD8-positivos/inmunología , Células Dendríticas/inmunología , Memoria Inmunológica , Orquitis/inmunología , Animales , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/patología , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Linfocitos T CD4-Positivos/inmunología , Comunicación Celular/inmunología , Enfermedad Crónica , Modelos Animales de Enfermedad , Humanos , Masculino , Ratones , Ratones Transgénicos , Orquitis/genética , Orquitis/patología , Proteínas Represoras/genética , Testículo/citología , Testículo/inmunología , Testículo/patologíaRESUMEN
Cgnz1 on chromosome 1 mapped into a 1.34 Mb region of chromosome 1 in NZM2328 confers the progression of immune complex (IC)-mediated glomerulonephritis (GN) from acute GN (aGN) to chronic GN (cGN) with severe proteinuria and end stage renal disease in female mice. This genetic locus mediates podocyte susceptibility to IC-mediated damage. Taking advantage of the published observation that Cgnz1 is derived from NZW and that NZW is susceptible to orchitis, epididymitis and vasitis while C57L/J is resistant to these diseases, the possibility that this genetic region also confers germ cells susceptible to damage with aspermatogenesis and sterility in an active experimental autoimmune orchitis (EAO) model was investigated. Male mice from multiple intrachromosome (chromosome 1) recombinant strains were subjected to immunization with a sperm homogenate in CFA with concomitant administration of Bordetella pertussis toxin. There was concordance of the progression from aGN to cGN, severe proteinuria and end stage renal disease with susceptibility of EAO in NZM2328 and its congenic strains with various chromosome 1 genetic intervals introgressed from C57L/J to NZM2328. Both resistant and susceptible strains made comparable anti-testis and anti-sperm Abs. Thus the genetic interval that determines susceptibility to EAO is identical to that of Cgnz1 and mapped to the 1.34 Mb region in chromosone 1. This region likely confers germ cells in the male gonad susceptible to damage by immunologically mediated inflammation. This region has been tentatively renamed Cgnz1/Eaoz1. These observations further emphasize the importance of end organ susceptibility to damage in the pathogenesis of both systemic and organ specific autoimmune diseases.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Predisposición Genética a la Enfermedad , Glomerulonefritis/inmunología , Fallo Renal Crónico/inmunología , Orquitis/inmunología , Animales , Enfermedades Autoinmunes/etiología , Enfermedades Autoinmunes/genética , Femenino , Regulación de la Expresión Génica/inmunología , Glomerulonefritis/complicaciones , Glomerulonefritis/genética , Fallo Renal Crónico/complicaciones , Fallo Renal Crónico/genética , Masculino , Ratones , Orquitis/etiología , Orquitis/genéticaRESUMEN
Inflammation is part of a defense reaction of live tissues that is triggered by pathogens, chemical reagents, trauma, and radiation. Understanding the inflammatory process triggered by Zika virus (ZIKV) is important to better understand the pathogen-host interaction. The evaluation of this process can be done using tools such as enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription PCR (RT-qPCR). Both techniques have been an indispensable tool not just for immunologists but for all interested in understanding the inflammatory process.
Asunto(s)
Inflamación/diagnóstico , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Virus Zika/fisiología , Animales , Barrera Hematotesticular/inmunología , Barrera Hematotesticular/metabolismo , Barrera Hematotesticular/virología , Muerte Celular , Ensayo de Inmunoadsorción Enzimática/métodos , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Humanos , Inflamación/genética , Inflamación/inmunología , Inflamación/virología , Masculino , Ratones , Orquitis/diagnóstico , Orquitis/genética , Orquitis/inmunología , Orquitis/virología , Testículo/patología , Testículo/fisiología , Virus Zika/inmunología , Virus Zika/patogenicidad , Infección por el Virus Zika/complicaciones , Infección por el Virus Zika/genética , Infección por el Virus Zika/inmunología , Infección por el Virus Zika/metabolismoRESUMEN
Fluoride (F) widely exists in the water and food. Recent studies reported that F induced testicular toxicity via inflammation reaction. This study was aimed to explore the mechanism of F-induced inflammation in testis. 100 healthy male mice (BALB/cJ strain) were randomly divided into five groups including: control, experimental autoimmune orchitis (EAO), and three F groups (25, 50, and 100 mg/L sodium fluoride (NaF)). After 150 d, the results showed a significant increase in testicular cytokines levels including of IL-17A, IL-6, IFN-γ, and TNF-α in NaF and EAO groups compared with control group. Interestingly, the presence of specific antisperm autoantibodies in antitesticular autoantibodies and the notable recruitment of immunocyte (T cells and dendritic cells) were also observed in NaF and EAO groups. In addition, findings showed that in NaF and EAO groups macrophages and T cells both significantly secreted IL-17A, and the protein and mRNA levels of cytokines (IL-6 and TGF-ß) were significantly increased. From these results, it can be concluded that autoimmune orchitis and IL-17A are implicated in F-induced testicular inflammation.
Asunto(s)
Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/inmunología , Fluoruros/efectos adversos , Interleucina-17/inmunología , Orquitis/inmunología , Testículo/inmunología , Animales , Enfermedades Autoinmunes/genética , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Humanos , Interleucina-17/genética , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Masculino , Ratones , Ratones Endogámicos BALB C , Orquitis/inducido químicamente , Orquitis/genética , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Testículo/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
STUDY QUESTION: What is the underlying mechanism of Sertoli cell (SC) resistance to cell death? SUMMARY ANSWER: High expression of prosurvival B-cell lymphoma-2 (BCL2) proteins and inhibition of apoptosis and autophagy prolongs SC survival upon exposure to stress stimuli. WHAT IS KNOWN ALREADY: In human and in experimental models of orchitis, tolerogenic SC survive stress conditions, while germ cells undergo massive apoptosis. In general, non-dividing highly differentiated cells tend to resist stress conditions for a longer time by favoring activation of prosurvival mechanisms and inhibition of cell death pathways. STUDY DESIGN, SIZE, DURATION: In this cross sectional study, conditions stimulating apoptosis and autophagy were used to induce cell death in primary rat SC. Primary rat peritubular cells (PTC) and immortalized rat 93RS2 SC were used as controls. Each cell isolation was counted as one experiment (n = 1), and each experiment was repeated three to six times. PARTICIPANTS/MATERIALS, SETTING, METHODS: Testis biopsy samples from infertile or subfertile patients and testis samples from rats with experimental autoimmune orchitis were used for immunohistological analysis. Primary SC were isolated from 19-day-old male Wistar rats. To maintain cell purity, cells were cultured in serum-free medium for apoptosis experiments and in medium supplemented with 1% serum for autophagy analyses. To induce apoptosis, cells were stimulated with staurosporine, borrelidin, cisplatin and etoposide for 4 or 24 h. Caspase three activation was examined by immunoblotting and enzymatic activity assay. Mitochondrial membrane potential was measured using tetramethylrhodamine methyl ester followed by flow cytometric analysis. Cytochrome c release was monitored by immunofluorescence. Cell viability was determined using the methylthiazole tetrazolium assay. To monitor autophagy flux, cells were deprived of nutrients using Hank's balanced salt solution for 1, 2 and 3 h. Formation of autophagosomes was analyzed by using immunoblotting, immunofluorescence labeling and ultrastructural analyses. Relative mRNA levels of genes involved in the regulation of apoptosis and autophagy were evaluated. Extracellular high mobility group box protein one was measured as a marker of necrosis using ELISA. MAIN RESULTS AND THE ROLE OF CHANCE: SC survive the inflammatory conditions in vivo in human testis and in experimental autoimmune orchitis. Treatment with apoptosis inducing chemotherapeutics did not cause caspase three activation in isolated rat SC. Moreover, mitochondrial membrane potential and mitochondrial localization of cytochrome c were not changed by treatment with staurosporine, suggesting a premitochondrial blockade of apoptosis in SC. Expression levels of prosurvival BCL2 family members were significantly higher in SC compared to PTC at both mRNA and protein levels. Furthermore, after nutrient starvation, autophagy signaling was initiated in SC as observed by decreased levels of phosphorylated UNC- 51-like kinase -1 (ULK1). However, levels of light chain 3 II (LC3 II) and sequestosome1 (SQSTM1) remained unchanged, indicating blockade of the autophagy flux. Lysosomal activity was intact in SC as shown by accumulation of LC3 II following administration of lysosomal protease inhibitors, indicating that inhibition of autophagy flux occurs at a preceding stage. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: In this study, we have used primary SC from prepubertal rats. Caution should be taken when translating our results to adult animals, where crosstalk with other testicular cells and hormonal factors may also play a role in regulating survival of SC. WIDER IMPLICATIONS OF THE FINDINGS: Our results suggest that inhibition of autophagy and apoptosis following exposure to extrinsic stress stimuli promotes SC survival, and is a possible mechanism to explain the robustness of SC in response to stress. Cell death resistance in SC is crucial for the recovery of spermatogenesis after chemotherapy treatment in cancer patients. Additionally, understanding the molecular mechanisms of SC survival unravels valuable target proteins, such as BCL2, that may be manipulated therapeutically to control cell viability depending on the context of the disease. STUDY FUNDING AND COMPETING INTEREST(S): This study was funded by the Deutsche Forschungsgemeinschaft (DFG) Grant BH93/1-1, and by the International Research Training Group between Justus Liebig University of Giessen and Monash University, Melbourne (GRK 1871/1) funded by the DFG and Monash University. The support of the Medical Faculty of Justus-Liebig University of Giessen is gratefully acknowledged. The authors declare no conflict of interest.
Asunto(s)
Proteínas Reguladoras de la Apoptosis/genética , Infertilidad Masculina/genética , Orquitis/genética , Proteínas Proto-Oncogénicas c-bcl-2/genética , Células de Sertoli/metabolismo , Animales , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/metabolismo , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/patología , Autofagia/efectos de los fármacos , Caspasa 3/genética , Caspasa 3/metabolismo , Supervivencia Celular/genética , Cisplatino/farmacología , Estudios Transversales , Citocromos c/metabolismo , Modelos Animales de Enfermedad , Etopósido/farmacología , Alcoholes Grasos/farmacología , Regulación del Desarrollo de la Expresión Génica , Humanos , Infertilidad Masculina/metabolismo , Infertilidad Masculina/patología , Masculino , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Orquitis/inmunología , Orquitis/patología , Cultivo Primario de Células , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Ratas Wistar , Células de Sertoli/efectos de los fármacos , Células de Sertoli/patología , Espermatogénesis/genética , Estaurosporina/farmacologíaRESUMEN
Although TiO2 nanoparticles (NPs) as endocrine disruptors have been demonstrated to be able to cross the blood-testis barriers and induce reproductive toxicity in male animals, whether the reproductive toxicity of male animals due to exposure to endocrine disruptor TiO2 NPs is related to immunological dysfunction in the testis remains not well understood. This study determined whether the reproductive toxicity and immunological dysfunction induced by exposure to TiO2 NPs is associated with activation or inhibition of TAM/TLR-mediated signal pathway in mouse testis. The results showed that male mice exhibited significant reduction of fertility, infiltration of inflammatory cells, rarefaction, apoptosis, and/or necrosis of spermatogenic cells and Sertoli cells due to TiO2 NPs. Furthermore, these were associated with decreased expression of Tyro3 (-18.16 to -66.6%), Axl (-14.7 to -57.99%), Mer (-7.98 to -72.62%), and IκB (-11.25 to -63.16%), suppression of cytokine signaling (SOCS) 1 (-21.99 to -73.8%) and SOCS3 (-8.11 to -34.86%), and increased expression of Toll-like receptor (TLR)-3 (21.4-156.03%), TLR-4 (37.0-109.87%), nuclear factor-κB (14.75-69.34%), interleukin (IL)-lß (46.15-123.08%), IL-6 (2.54-81.98%), tumor necrosis factor-α (6.95-88.39%), interferon (IFN)-α (2.54-37.25%), and IFN-ß (10.19-80.56%), which are involved in the immune environment in the testis. The findings showed that reproductive toxicity of male mice induced by exposure to endocrine disruptor TiO2 NPs may be associated with biomarkers of impairment of immune environment or dysfunction of TAM/TLR3-mediated signal pathway in mouse testitis. Therefore, the potential risks to reproductive health should be attended, especially in those who are occupationally exposed to TiO2 NPs.
Asunto(s)
Nanopartículas/toxicidad , Orquitis/inmunología , Titanio/toxicidad , Animales , Humanos , Masculino , Ratones , Ratones Endogámicos ICR , Orquitis/etiología , Orquitis/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/inmunología , Proteínas Tirosina Quinasas Receptoras/genética , Proteínas Tirosina Quinasas Receptoras/inmunología , Proteína 1 Supresora de la Señalización de Citocinas , Proteínas Supresoras de la Señalización de Citocinas/genética , Proteínas Supresoras de la Señalización de Citocinas/inmunología , Testículo/efectos de los fármacos , Testículo/inmunología , Tirosina Quinasa del Receptor AxlRESUMEN
Galectin-1 (Gal-1), a proto-type member of galectin family, is highly expressed in immune privileged sites, including the testis. However, in spite of considerable progress the relevance of endogenous and exogenous Gal-1 in testis pathophysiology have not yet been explored. Here we evaluated the in vivo roles of Gal-1 in experimental autoimmune orchitis (EAO), a well-established model of autoimmune testicular inflammation associated with subfertility and infertility. A significant reduction in the incidence and severity of EAO was observed in mice genetically deficient in Gal-1 (Lgals1(-/-)) versus wild-type (WT) mice. Testicular histopathology revealed the presence of multifocal testicular damage in WT mice characterized by an interstitial mononuclear cell infiltrate and different degrees of germ cell sloughing of seminiferous tubules. TUNEL assay and assessment of active caspase-3 expression, revealed the prevalence of apoptotic spermatocytes mainly localized in the adluminal compartment of seminiferous tubules in EAO mice. A significant increased number of TUNEL-positive germ cells was detected in EAO testis from WT compared with Lgals1(-/-) mice. In contrast, exogenous administration of recombinant Gal-1 to WT mice undergoing EAO attenuated the severity of the disease. Our results unveil a dual role of endogenous versus exogenous Gal-1 in the control of autoimmune testis inflammation.
Asunto(s)
Apoptosis/inmunología , Enfermedades Autoinmunes/inmunología , Galectina 1/inmunología , Orquitis/inmunología , Túbulos Seminíferos/inmunología , Espermatocitos/inmunología , Animales , Apoptosis/genética , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/patología , Galectina 1/genética , Masculino , Ratones , Ratones Noqueados , Orquitis/genética , Orquitis/patología , Túbulos Seminíferos/patología , Espermatocitos/patologíaRESUMEN
Gap junction-mediated communication helps synchronize interconnected Sertoli cell activities. Besides, coordination of germ cell and Sertoli cell activities depends on gap junction-mediated Sertoli cell-germ cell communication. This report assesses mechanisms underlying the regulation of connexin 46 (Cx46) and Cx50 in mouse testis and those accompanying a "natural" seasonal and a pathological arrest of spermatogenesis, resulting from autoimmune orchitis (AIO) in mink. Furthermore, the impact of deleting Cx46 or Cx50 on the expression, phosphorylation of junction proteins, and spermatogenesis is evaluated. Cx46 mRNA and protein expression increased, whereas Cx50 decreased with adulthood in normal mice and mink. Cx46 mRNA and protein expression increased, whereas Cx50 decreased with adulthood in normal mice and mink. During the mink active spermatogenic phase, Cx50 became phosphorylated and localized to the site of the blood-testis barrier. By contrast, Cx46 was dephosphorylated and associated with annular junctions, suggesting phosphorylation/dephosphorylation of Cx46 and Cx50 involvement in the barrier dynamics. Cx46-positive annular junctions in contact with lipid droplets were found. Cx46 and Cx50 expression and localization were altered in mink with AIO. The deletion of Cx46 or Cx50 impacted on other connexin expression and phosphorylation and differently affected tight and adhering junction protein expression. The level of apoptosis, determined by ELISA, and a number of Apostain-labeled spermatocytes and spermatids/tubules were higher in mice lacking Cx46 (Cx46-/-) than wild-type and Cx50-/- mice, arguing for life-sustaining Cx46 gap junction-mediated exchanges in late-stage germ cells secluded from the blood by the barrier. The data show that expression and phosphorylation of Cx46 and Cx50 are complementary in seminiferous tubules.
Asunto(s)
Comunicación Celular/fisiología , Conexinas/metabolismo , Eliminación de Gen , Orquitis/metabolismo , Testículo/metabolismo , Animales , Conexinas/genética , Uniones Comunicantes/metabolismo , Cristalino , Masculino , Ratones Endogámicos BALB C , Visón , Orquitis/genética , FosforilaciónRESUMEN
The mammalian testis is an immunoprivileged organ where male germ cell autoantigens are immunologically ignored. Both systemic immune tolerance to autoantigens and local immunosuppressive milieu contribute to the testicular immune privilege. Testicular immunosuppression has been intensively studied, but information on systemic immune tolerance to autoantigens is lacking. In the present study, we aimed to determine the role of Axl and Mer receptor tyrosine kinases in maintaining the systemic tolerance to male germ cell antigens using the experimental autoimmune orchitis (EAO) model. Axl and Mer double-knockout (Axl(-/-)Mer(-/-)) mice developed evident EAO after a single immunization with germ cell homogenates emulsified with complete Freund's adjuvant. EAO was characterized by the accumulation of macrophages and T lymphocytes in the testis. Damage to the seminiferous epithelium was also observed. EAO induction was associated with pro-inflammatory cytokine upregulation in the testes, impaired permeability of the blood-testis barrier and generation of autoantibodies against germ cell antigens in Axl(-/-)Mer(-/-) mice. Immunization also induced mild EAO in Axl or Mer single-gene-knockout mice. By contrast, a single immunization failed to induce EAO in wild-type mice. The results indicate that Axl and Mer receptors cooperatively regulate the systemic immune tolerance to male germ cell antigens.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Macrófagos/inmunología , Orquitis/inmunología , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Tirosina Quinasas Receptoras/metabolismo , Linfocitos T/inmunología , Testículo/inmunología , Animales , Autoanticuerpos/inmunología , Autoantígenos/inmunología , Enfermedades Autoinmunes/genética , Barrera Hematotesticular/inmunología , Modelos Animales de Enfermedad , Humanos , Tolerancia Inmunológica , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Orquitis/genética , Proteínas Proto-Oncogénicas/genética , Proteínas Tirosina Quinasas Receptoras/genética , Tirosina Quinasa c-Mer , Tirosina Quinasa del Receptor AxlRESUMEN
First reported as a vasoactive peptide in the cardiovascular system, intermedin (IMD), also known as adrenomedullin 2 (ADM2), is a hormone with multiple potent roles, including its antioxidant action on the pulmonary, central nervous, cardiovascular and renal systems. Though IMD may play certain roles in trophoblast cell invasion, early embryonic development and cumulus cell-oocyte interaction, the role of IMD in the male reproductive system has yet to be investigated. This paper reports our findings on the gene expression of IMD, its receptor components and its protein localization in the testes. In a rat model, bacterial lippolysaccharide (LPS) induced atypical orchitis, and LPS treatment upregulated the expression of IMD and one of its receptor component proteins, i.e. receptor activity modifying protein 2 (RAMP2). IMD decreased both plasma and testicular levels of reactive oxygen species (ROS) production, attenuated the increase in the gene expression of the proinflammatory cytokines tumor necrosis factor alpha (TNFα), interleukin 6 (IL6) and interleukin 1 beta (IL1ß), rescued spermatogenesis, and prevented the decrease in plasma testosterone levels caused by LPS. The restorative effect of IMD on steroidogenesis was also observed in hydrogen peroxide-treated rat primary Leydig cells culture. Our results indicate IMD plays an important protective role in spermatogenesis and steroidogenesis, suggesting therapeutic potential for IMD in pathological conditions such as orchitis.
Asunto(s)
Adrenomedulina/metabolismo , Lipopolisacáridos/farmacología , Orquitis/inducido químicamente , Orquitis/metabolismo , Testículo/metabolismo , Adrenomedulina/genética , Animales , Peso Corporal/efectos de los fármacos , Citocinas/genética , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/farmacología , Inflamación/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Orquitis/genética , Estrés Oxidativo/efectos de los fármacos , Transporte de Proteínas/efectos de los fármacos , Ratas , Receptores de Adrenomedulina/metabolismo , Testículo/efectos de los fármacos , Testosterona/sangre , Testosterona/metabolismoRESUMEN
Lymphangiogenesis occurs in various organs under inflammatory conditions. Recently, it was demonstrated that activated macrophages play an important role in the process of lymphangiogenesis. However, lymphangiogenesis during testicular inflammation has not yet been studied. Here, we investigated lymphangiogenesis in experimental autoimmune orchitis, a immunologic male infertility model, in mice. Histological changes were observed using immunohistochemical staining with the monoclonal antibodies against F4/80 (mature macrophage marker), lymph vessel endothelium HA-receptor 1 (LYVE-1) (lymphatic endothelial cells marker) and CD31 (endothelial cells marker). The expression of angiogenesis and lymphangiogenesis factors, such as vascular endothelial growth factor (VEGF)-A, VEGF-C, VEGF-D and TNF-α, which are secreted by activated macrophages, were examined using real-time RT-PCR. The results showed that lymphangiogenesis occurred along the undersurface of the tunica albuginea but not into the interstitium proper between the seminiferous tubules (STs) during the orchitis. It was noted that some F4/80-positive macrophages expressed LYVE-1 at the undersurface of the tunica albuginea and also in the testicular interstitium proper. RT-PCR analysis revealed that the expressions of VEGF-A, VEGF-D and TNF-α were significantly increased but that of VEGF-C remained unchanged in the inflammatory testes. This study suggests that testicular macrophages are involved in the specific lymphangiogenesis in the chronic inflammation.
Asunto(s)
Linfangiogénesis , Vasos Linfáticos/fisiopatología , Testículo/fisiopatología , Animales , Antígenos de Diferenciación/metabolismo , Biomarcadores/metabolismo , Enfermedad Crónica , Modelos Animales de Enfermedad , Glicoproteínas/metabolismo , Inmunohistoquímica , Vasos Linfáticos/metabolismo , Vasos Linfáticos/patología , Activación de Macrófagos , Macrófagos/metabolismo , Macrófagos/patología , Masculino , Proteínas de Transporte de Membrana , Ratones , Orquitis/genética , Orquitis/metabolismo , Orquitis/patología , Orquitis/fisiopatología , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Testículo/metabolismo , Testículo/patología , Factores de Tiempo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor C de Crecimiento Endotelial Vascular/genética , Factor C de Crecimiento Endotelial Vascular/metabolismo , Factor D de Crecimiento Endotelial Vascular/genética , Factor D de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Experimental autoimmune orchitis (EAO) is a model of immunologic male infertility and pathologically characterized by lymphocytic inflammation, which causes breakdown of the testicular immune privilege with spermatogenic disturbance. Generally, murine EAO is induced by immunization with testicular homogenate (TH) from the testes of donor mice + complete Freund's adjuvant (CFA) + Bordetella pertussigens (BP), and it has been considered that treatment with these two adjuvants is required to enhance the immune response against testicular antigens. However, there remains a possibility that CFA and BP may affect autoimmune responses against the testicular antigens without TH. In the present study, we examined this possibility using real-time RT-PCR, Western blotting and immunohistochemical staining. The results demonstrated that immunization with TH in combination with CFA and BP evoked more severe EAO than that with only TH. Real-time RT-PCR analyses revealed that Fas mRNA expression in TH+CFA+BP-induced EAO was significantly higher than that in TH-induced EAO. Interestingly, IL-6 mRNA expression dramatically increased in TH+CFA+BP-induced EAO; however, no apparent change in IL-6 mRNA expression occurred in TH-induced EAO. It was also noted that treatment with CFA and BP alone augmented autoimmune reactions against some testicular autoantigens. These results indicates that these adjuvants are helpful in evoking severe EAO, and treatment with the adjuvants alone can evoke autoimmune reactions against some testicular autoantigens despite the use of no TH.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Autoantígenos/farmacología , Enfermedades Autoinmunes/inmunología , Autoinmunidad/efectos de los fármacos , Infertilidad Masculina/inmunología , Orquitis/inmunología , Testículo/efectos de los fármacos , Animales , Autoantígenos/inmunología , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/metabolismo , Autoinmunidad/inmunología , Modelos Animales de Enfermedad , Infertilidad Masculina/genética , Infertilidad Masculina/metabolismo , Inflamación/genética , Inflamación/inmunología , Inflamación/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Ratones , Orquitis/genética , Orquitis/metabolismo , Testículo/inmunologíaRESUMEN
Experimental autoimmune orchitis (EAO) is characterized by T cell-dependent lymphocytic inflammation and seminiferous tubule damage, which can result in the death of germ cells. The aim of the present study is to investigate the roles of the Fas/Fas-L and Bax/Bcl-2 systems in the death of germ cells in mice with EAO that is induced by immunization with syngeneic testicular germ cells (TGC). The results using real-time reverse transcription-polymerase chain reaction and immunostaining show that many terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining germ cells were present in seminiferous tubules during the active inflammation stage, and these cells were persistently observed in the seminiferous epithelium until the postactive inflammation stage. Intratesticular mRNA expression levels of both Fas and Bax were increased during the active inflammation stage and were dramatically decreased during the post-active inflammation stage. In contrast, the intratesticular mRNA expression levels of both Fas-L and Bcl-2 did not show significant changes during the active inflammation stage but showed extreme increases during the post-active inflammation stage. Immunohistochemically, some Fas- and Bax-positive germ cells were detected during the active inflammation stage, but these were hardly found during the post-active inflammation stage. In contrast, some Fas-L- and Bcl-2-positive germ cells were found during the active inflammation stage, and many of these were also observed during the post-active inflammation stage. These results indicate that germ cell death during TGC-induced EAO is mediated by the Fas/Fas-L and Bax/Bcl-2 systems during the active inflammation stage but not during the post-active inflammation stage.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Proteína Ligando Fas/metabolismo , Orquitis/inmunología , Proteínas Proto-Oncogénicas/metabolismo , Espermatozoides/trasplante , Testículo/trasplante , Proteína X Asociada a bcl-2/metabolismo , Receptor fas/metabolismo , Animales , Apoptosis , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/metabolismo , Enfermedades Autoinmunes/patología , Autoinmunidad , Modelos Animales de Enfermedad , Proteína Ligando Fas/genética , Regulación de la Expresión Génica , Inmunización , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Masculino , Ratones , Orquitis/genética , Orquitis/metabolismo , Orquitis/patología , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas c-bcl-2 , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Túbulos Seminíferos/inmunología , Túbulos Seminíferos/metabolismo , Túbulos Seminíferos/patología , Transducción de Señal , Espermatozoides/inmunología , Espermatozoides/metabolismo , Espermatozoides/patología , Testículo/inmunología , Testículo/metabolismo , Testículo/patología , Factores de Tiempo , Proteína X Asociada a bcl-2/genética , Receptor fas/genéticaRESUMEN
Experimental autoimmune orchitis (EAO), the principal model of non-infectious testicular inflammatory disease, can be induced in susceptible mouse strains by immunization with autologous testicular homogenate and appropriate adjuvants. As previously established, the genome of DBA/2J mice encodes genes that are capable of conferring dominant resistance to EAO, while the genome of BALB/cByJ mice does not and they are therefore susceptible to EAO. In a genome scan, we previously identified Orch3 as the major quantitative trait locus controlling dominant resistance to EAO and mapped it to chromosome 11. Here, by utilizing a forward genetic approach, we identified kinesin family member 1C (Kif1c) as a positional candidate for Orch3 and, using a transgenic approach, demonstrated that Kif1c is Orch3. Mechanistically, we showed that the resistant Kif1c(D2) allele leads to a reduced antigen-specific T cell proliferative response as a consequence of decreased MHC class II expression by antigen presenting cells, and that the L(578) â P(578) and S(1027) â P(1027) polymorphisms distinguishing the BALB/cByJ and DBA/2J alleles, respectively, can play a role in transcriptional regulation. These findings may provide mechanistic insight into how polymorphism in other kinesins such as KIF21B and KIF5A influence susceptibility and resistance to human autoimmune diseases.
Asunto(s)
Resistencia a la Enfermedad/genética , Genes Dominantes , Cinesinas/genética , Orquitis , Alelos , Animales , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/inmunología , Expresión Génica , Genes MHC Clase II , Predisposición Genética a la Enfermedad , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos DBA , Orquitis/genética , Orquitis/inmunología , Sitios de Carácter Cuantitativo/genética , Testículo/inmunologíaRESUMEN
Structural polymorphisms (L263P, M313V, and S331P) in the third intracellular loop of the murine histamine receptor H(1) (H(1)R) are candidates for Bphs, a shared autoimmune disease locus in experimental allergic encephalomyelitis and experimental allergic orchitis. The P-V-P haplotype is associated with increased disease susceptibility (H(1)R(S)) whereas the L-M-S haplotype is associated with less severe disease (H(1)R(R)). In this study, we show that selective re-expression of the H(1)R(S) allele in T cells fully complements experimental allergic encephalomyelitis susceptibility and the production of disease-associated cytokines while selective re-expression of the H(1)R(R) allele does not. Mechanistically, we show that the two H(1)R alleles exhibit differential cell surface expression and altered intracellular trafficking, with the H(1)R(R) allele being retained within the endoplasmic reticulum. Moreover, we show that all three residues (L-M-S) comprising the H(1)R(R) haplotype are required for altered expression. These data are the first to demonstrate that structural polymorphisms influencing cell surface expression of a G protein-coupled receptor in T cells regulates immune functions and autoimmune disease susceptibility.
Asunto(s)
Enfermedades Autoinmunes/genética , Encefalomielitis Autoinmune Experimental/genética , Orquitis/genética , Receptores Histamínicos H1/genética , Alelos , Animales , Enfermedades Autoinmunes/inmunología , Línea Celular , Membrana Celular/metabolismo , Encefalomielitis Autoinmune Experimental/inmunología , Retículo Endoplásmico/metabolismo , Subunidades alfa de la Proteína de Unión al GTP/metabolismo , Predisposición Genética a la Enfermedad , Haplotipos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Orquitis/inmunología , Polimorfismo Genético , Transporte de Proteínas , Receptores Histamínicos H1/deficiencia , Receptores Histamínicos H1/inmunología , Receptores Histamínicos H1/metabolismo , Linfocitos T/metabolismoRESUMEN
Symptoms of tuberculous orchiepididymitis in a 39-year-old male started with swelling of left scrotum, followed by fistula formation with suppurative discharge. There was no any improvement produced by antibiotics. Surgical extirpation of inflammatory destroyed testicle and epidydimis was performed. Presence of tubercle bacilli was not shown by bacteriological analysis of testicle tissue. Tuberculous etiology was suggested after histopathological examination of testis and epididymis. Exudate from surgical wound was examined on presence of Mycobacterium tuberculosis DNA. Etiology of orchiepididymitis was proved by positive assay and inflammatory process was completely cured by antituberculotics therapy. By this report it was clearly shown that sometimes only molecular methods could confirm etiology of inflammatory process.
Asunto(s)
Epididimitis/diagnóstico , Epididimitis/microbiología , Orquitis/diagnóstico , Orquitis/microbiología , Tuberculosis de los Genitales Masculinos/diagnóstico , Adulto , Epididimitis/genética , Epididimitis/cirugía , Humanos , Masculino , Técnicas de Amplificación de Ácido Nucleico , Orquitis/genética , Orquitis/cirugía , Tuberculosis de los Genitales Masculinos/genética , Tuberculosis de los Genitales Masculinos/cirugíaRESUMEN
An important approach to understanding complex diseases is to reduce them into well-characterized subphenotypes that are under monogenic control. One such example is Bordetella pertussis toxin-induced histamine sensitization in mice, a subphenotype of experimental allergic encephalomyelitis and experimental allergic orchitis. This subphenotype is controlled by a single locus, Bphs, previously mapped to a 33 cM region on mouse Chromosome (Chr) 6. We achieved considerable reduction of this candidate region and constructed a YAC contig across the refined interval. Our results demonstrate that Bphs is located between D6Mit151 and a newly developed marker, EC108RR, a region containing a small cluster of genes belonging to the TNF receptor superfamily. Sequence and quantitative analysis of the candidate gene, tumor necrosis factor receptor 1 (Tnfr1, p55), indicates that it is unlikely to be Bphs. However, the location of Bphs, together with physiologic effects it shares with Tnfr1 activation, suggest that Bphs may prove to be another member of the TNF receptor superfamily.
Asunto(s)
Enfermedades Autoinmunes/genética , Familia de Multigenes , Receptores del Factor de Necrosis Tumoral/genética , Animales , Cromosomas Artificiales de Levadura/genética , Encefalomielitis Autoinmune Experimental/genética , Marcadores Genéticos , Liberación de Histamina/efectos de los fármacos , Liberación de Histamina/genética , Masculino , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos CBA , Orquitis/genética , Orquitis/inmunología , Toxina del Pertussis , Mapeo Físico de Cromosoma , Polimorfismo Genético , ARN Mensajero/análisis , ARN Mensajero/genética , Recombinación Genética , Factores de Virulencia de Bordetella/toxicidadRESUMEN
Experimental allergic orchitis (EAO), the principle animal model of noninfectious testicular inflammatory disease, is a genetically determined phenotype. Classical EAO, induced by inoculation with testicular homogenate and the appropriate adjuvants, is characterized by inflammatory infiltrates in the testis (orchitis), epididymis (epididymitis), and vas deferens (vasitis). In this study, the genetic control of susceptibility and resistance to these three lesions was analyzed in the mouse. The results obtained with independent inbred strains and H2 congenic mice show that the genetic control of all three lesions is complex and involves both H2 and non-H2-linked genes. Whole-genome exclusion mapping was performed on a backcross population segregating for all three phenotypes. Permutation-derived thresholds provided experimentwise, chromosomewise, comparisonwise, and marker-specific chromosomewise thresholds for declaration of significant regions linked to marker loci. Unique loci were identified on chromosome 8 for orchitis, chromosome 16 for epididymitis, and chromosome 1 for vasitis and have been designated as Orch6, Epd1, and Vas1, respectively. These results show that autoimmune orchitis, epididymitis, and vasitis are immunogenetically distinct lesions.
Asunto(s)
Enfermedades Autoinmunes/genética , Epididimitis/genética , Orquitis/genética , Conducto Deferente/patología , Animales , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/patología , Mapeo Cromosómico , ADN/análisis , Susceptibilidad a Enfermedades/inmunología , Susceptibilidad a Enfermedades/patología , Epididimitis/inmunología , Epididimitis/patología , Amplificación de Genes , Ligamiento Genético , Predisposición Genética a la Enfermedad , Genoma , Genotipo , Antígenos H-2/genética , Endogamia , Masculino , Ratones , Ratones Endogámicos , Orquitis/inmunología , Orquitis/patología , Polimorfismo Genético , Conducto Deferente/inmunologíaRESUMEN
Previous studies have shown that differential susceptibility to actively induced experimental allergic encephalomyelitis (EAE) and experimental allergic orchitis (EAO) exists among various BALB/c substrains. Of eight substrains studied for EAE and 13 for EAO, BALB/cJ mice are phenotypically the most resistant to disease induction. Resistance to both diseases is controlled by single recessive mutations unlinked to any of the known alleles distinguishing BALB/cJ mice. In this study, segregation analysis employing a second generation backcross population shows that resistance to both EAE and EAO is due to a mutation in a common immunoregulatory gene. The role of immunoregulatory cells in controlling EAE resistance was examined using adoptive transfer protocols. BALB/cJ mice immunized with spinal cord homogenate plus adjuvants generate immunoregulatory spleen cells (SpC) that, when transferred to naive BALB/cByJ recipients, reduce the incidence and severity of EAE. Treatment of such cells with either cytotoxic monoclonal anti-Thy1.2 or anti-CD4 plus C' before transfer abrogates the ability of BALB/cJ SpC to inhibit disease. In contrast, neither SpC from adjuvant-immunized BALB/cJ nor spinal cord homogenate- plus adjuvant-primed BALB/cByJ donors influences the incidence or severity of disease observed in recipients. In addition, the role of environment in influencing susceptibility to EAE and EAO in BALB/c mice is documented. Taken together, these results support the existence of a common disease susceptibility locus in the pathways leading to two autoantigenically distinct CD4+ T cell-mediated, organ-specific, autoimmune diseases.
Asunto(s)
Enfermedades Autoinmunes/genética , Encefalomielitis Autoinmune Experimental/genética , Predisposición Genética a la Enfermedad , Orquitis/genética , Animales , Enfermedades Autoinmunes/inmunología , Linfocitos T CD4-Positivos/inmunología , Encefalomielitis Autoinmune Experimental/inmunología , Femenino , Masculino , Ratones , Ratones Endogámicos BALB C , Mutación , Orquitis/inmunología , Especificidad de ÓrganosRESUMEN
A wide range of autoimmune and other diseases are known to be associated with the major histocompatibility complex. Many of these diseases are linked to the genes encoding the polymorphic histocompatibility antigens in the class I and class II regions, but some appear to be more strongly associated with genes in the central 1100-kb class III region, making it important to characterize this region fully for the presence of novel genes. An approximately 220-kb segment of DNA in the class III region separating the Hsp70 (HSPA1L) and BAT1 (D6S8IE) genes, which was previously known to contain 14 genes, has been analyzed for the presence of additional genes. Genomic DNA fragments spanning the gaps between the known genes were used as probes to isolate cDNAs corresponding to five new genes within this region. Evidence from Northern blot analysis and exon trapping experiments that suggested the presence of at least two more new genes was also obtained. Partial cDNA and complete exonic genomic sequencing of one of the new genes has identified it as the casein kinase II beta subunit (CSNK2B). Two of the other novel genes lie within a region syntenic to that implicated in susceptibility to experimental allergic orchitis in the mouse, an autoimmune disease of the testis, and represent additional candidates for the Orch-1 locus associated with this disease. In addition, characterization of the 13-kb intergenic gap separating the RD (D6545) and G11 (D6S60E) genes has revealed the presence of a gene encoding a 1246-amino-acid polypeptide that shows significant sequence similarity to the yeast anti-viral Ski2p gene product.