RESUMEN
SAMP1/YitFcs mice serve as a model of Crohn's disease, and we have used them to assess gastritis. Gastritis was compared in SAMP1/YitFcs, AKR, and C57BL/6 mice by histology, immunohistochemistry, and flow cytometry. Gastric acid secretion was measured in ligated stomachs, while anti-parietal cell antibodies were assayed by immunofluorescence and enzyme-linked immunosorbent spot assay. SAMP1/YitFcs mice display a corpus-dominant, chronic gastritis with multifocal aggregates of mononuclear cells consisting of T and B lymphocytes. Relatively few aggregates were observed elsewhere in the stomach. The infiltrates in the oxyntic mucosa were associated with the loss of parietal cell mass. AKR mice, the founder strain of the SAMP1/YitFcs, also have gastritis, although they do not develop ileitis. Genetic studies using SAMP1/YitFcs-C57BL/6 congenic mice showed that the genetic regions regulating ileitis had comparable effects on gastritis. The majority of the cells in the aggregates expressed the T cell marker CD3 or the B cell marker B220. Adoptive transfer of SAMP1/YitFcs CD4(+) T helper cells, with or without B cells, into immunodeficient recipients induced a pangastritis and duodenitis. SAMP1/YitFcs and AKR mice manifest hypochlorhydria and anti-parietal cell antibodies. These data suggest that common genetic factors controlling gastroenteric disease in SAMP1/YitFcs mice regulate distinct pathogenic mechanisms causing inflammation in separate sites within the digestive tract.
Asunto(s)
Aclorhidria/inmunología , Enfermedades Autoinmunes/inmunología , Gastritis/inmunología , Ileítis/inmunología , Aclorhidria/genética , Aclorhidria/patología , Traslado Adoptivo , Animales , Autoanticuerpos/análisis , Autoanticuerpos/inmunología , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/patología , Linfocitos B/inmunología , Linfocitos B/patología , Complejo CD3/análisis , Complejo CD3/inmunología , Femenino , Ácido Gástrico/metabolismo , Gastritis/genética , Gastritis/patología , Ileítis/genética , Ileítis/patología , Antígenos Comunes de Leucocito/análisis , Antígenos Comunes de Leucocito/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Linfocitos T/inmunología , Linfocitos T/patología , Linfocitos T Colaboradores-Inductores/inmunologíaRESUMEN
We describe a unique spontaneous mouse model of autoimmunity, which occurs on a non-autoimmune-prone SWR genetic background. In this model, SWR mice carry an IghV partial transgene (pTg) encoding only the heavy chain variable domain of an antibody directed against chromatin. Autoimmune disease in pTg mice was manifested by some of the features of systemic lupus erythematosus (SLE), including the presence of serum anti-nuclear antibodies, splenomegaly, skin lesions and a moderate degree of kidney pathology, in various combinations among individuals. Autoimmunity was observed in three independent transgenic lines, but not in three control lines carrying a nearly identical pTg, in which a VHCDR3 codon for Arg was replaced by one for Ser to ablate chromatin reactivity. Various features of disease were often but not always accompanied by anti-chromatin antibodies. Unexpectedly, the anti-chromatin antibodies detected in seropositive animals were not encoded by the pTg. These observations strongly implicate a role for the transgene product in disease initiation but not necessarily for end-state pathology, and they raise the possibility that autoreactive B cells may play a previously unappreciated role in initiating the development of systemic autoimmunity.
Asunto(s)
Autoinmunidad/inmunología , Modelos Animales de Enfermedad , Lupus Eritematoso Sistémico/inmunología , Animales , Anticuerpos Antinucleares/inmunología , Arginina/genética , Autoinmunidad/genética , Linfocitos B/inmunología , Cromatina/inmunología , Riñón/inmunología , Riñón/patología , Ratones , Ratones Transgénicos , Esplenomegalia/inmunología , TransgenesRESUMEN
Discovery of the CD4+CD25+ T cells has stemmed from investigation of the AOD in the d3tx mice. Besides CD4+CD25+ T cell depletion, d3tx disease induction requires effector T cell activation prompted by lymphopenia. This is supported by other neonatal AOD models in which T cell-mediated injury has been found to be triggered by immune complex or Ag immunization. In addition, there is growing evidence that support a state of neonatal propensity to autoimmunity, which depends on concomitant endogenous antigenic stimulation, concomitant nematode infection, resistance to CD4+CD25+ T cell regulation, and participation of the neonatal innate system. The suppression of d3tx disease by polyclonal CD4+CD25+ T cells appears to be dependent on endogenous Ag and the persistence of regulatory T cells. Thus, suppression of AOD occurs in the ovarian LN, and AOD emerges upon ablation of the input regulatory T cells; and in AIP, the hormone-induced expression of prostate Ag in the CD4+CD25+ T cell donors rapidly enhances the capacity to suppress disease over Ag negative donors. Finally, genetic analysis of AOD and its component phenotypes has uncovered seven Aod loci. As the general themes that emerged, significant epistatic interactions among the loci play a role in controlling disease susceptibility, the majority of the Aod loci are linked to susceptibility loci of other autoimmune diseases, and the genetic intervals encompass candidate genes that are differentially expressed between CD4+CD25+ T cells and other T cells. The candidate genes include Pdcd1, TNFR superfamily genes, H2, Il2, Tgfb, Nalp5 or Mater, an oocyte autoAg that reacts with autoantibody in sera of d3tx mice.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Enfermedades del Ovario/inmunología , Subgrupos de Linfocitos T/inmunología , Linfocitos T/inmunología , Animales , Antígenos/inmunología , Enfermedades Autoinmunes/patología , Linfocitos T CD4-Positivos/inmunología , Femenino , Predisposición Genética a la Enfermedad , Tolerancia Inmunológica , Ratones , Enfermedades del Ovario/patología , Receptores de Interleucina-2/inmunología , Especificidad del Receptor de Antígeno de Linfocitos T , TimectomíaRESUMEN
Recent studies on autoimmune ovarian disease (AOD) induced by thymectomy on d3 (d3tx), and AOD induced by immunization with the ovary-specific zona pellucida 3 peptide (pZP3), have yielded the following results. First, female tolerance to pZP3 depends on the persistence of endogenous antigen (Ag). Second, following regulatory T-cell depletion, endogenous Ag in prepubertal d3tx mice triggers AOD and drives disease progression. Third, endogenous ZP3 from ovaries without AOD stimulates a diversified IgG autoantibody (autoAb) response that rapidly follows pZP3 T epitope immunization. Fourth, induction of AOD and autoimmune memory in neonatal female mice by pZP3 in incomplete Freund's adjuvant depends on endogenous Ag stimulation within the neonatal week. Fifth, in a rodent pinworm-positive environment, neonatal but not adult female mice injected with pZP3 in water develop Th2-mediated AOD and Th2 memory. Sixth, neonatal T cells transfer AOD to syngeneic athymic recipients, whereas adult T cells are non-pathogenic and in fact suppress AOD conferred by neonatal T cells. Therefore: 1) the continuous presence of physiologically-expressed autoAg is critical for both tolerance maintenance and autoimmune disease pathogenesis; the outcome is determined by the integrity of regulatory T cells; and 2) the neonatal mice, deficient in the regulatory T-cell function, are more responsive than adults to Ag and environmental stimuli that promote autoimmune disease and memory.
Asunto(s)
Animales Recién Nacidos/inmunología , Autoantígenos/inmunología , Enfermedades Autoinmunes/inmunología , Enfermedades del Ovario/inmunología , Linfocitos T/inmunología , Animales , Animales Recién Nacidos/metabolismo , Autoantígenos/metabolismo , Enfermedades Autoinmunes/metabolismo , Ambiente , Femenino , Masculino , Enfermedades del Ovario/metabolismo , Ovario/inmunología , Ovario/metabolismo , Timectomía , Timo/citología , Timo/inmunologíaRESUMEN
Among NZB/W-derived New Zealand mixed (NZM) strains, only NZM/Aeg2410 (NZM2410) has been well characterized. In contrast to NZM2410, NZM2328 mice develop autoantibodies and acute and severe chronic glomerulonephritis (GN) with female predominance similarly to NZB/WF1 and humans with systemic lupus erythematosus (SLE). Chronic GN with glomerular sclerosis and tubular atrophy but not acute GN was correlated with severe proteinuria. In a backcross analysis of (NZM2328 X C57L/J) F1 X NZM2328, four SLE susceptibility genomic intervals were identified. One of them (Cgnz1) is on the telomeric end of chromosome 1 and close to Sle1. It was significantly linked to chronic GN. A locus (Agnz1) distinct from Cgnz1 on this interval was suggestively linked to acute GN. Two genetic intervals on chromosome 17 were also suggestively linked to acute GN, one of which is the H-2-Tnf complex, while the other (Agnz2) is on the distal end of the chromosome. A single locus (Adaz1) identified in the midregion of chromosome 4 in NZM2328 mice was suggestively linked to plasma levels of IgG anti-dsDNA autoantibodies. These results differ significantly from those in the backcross analysis of (NZM2410 X C57BL/6)F1 X NZM2410 by other investigators. They support the concept that different sets of genes are involved in acute and chronic GN. The genomic differences between the NZM strains and between C57L/J and C57BL/6 account for the differences between our analysis and that on NZM 2410. These results provide evidence for the importance of background genes on the expression of SLE, with implications for genetic studies of human SLE.
Asunto(s)
Mapeo Cromosómico , Modelos Animales de Enfermedad , Predisposición Genética a la Enfermedad , Lupus Eritematoso Sistémico/genética , Animales , Anticuerpos Antinucleares/sangre , ADN/inmunología , Ligamiento Genético , Genotipo , Glomerulonefritis/genética , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos NZBRESUMEN
Genetic and environmental factors both influence autoimmune disease occurrence, but the identity and mechanism of action of environmental factors are poorly understood. Here we show that pinworm-infected neonatal but not adult mice, injected with an ovarian self peptide of the zona pellucida protein 3 (pZP3) in water and without adjuvant, develop Th2 responses and severe eosinophilic autoimmune ovarian disease. A strong Th2 memory response is recalled when, as adults, the mice are challenged with a regimen that elicits a strong Th1 response in naive adults. The strong Th2 autoimmune response included high levels of IL-4 and IL-5 production by pZP3-specific T cells, and an IgG1-biased autoantibody response. The Th2 response ended promptly upon pinworm eradication, and partially resurfaced upon re-infection. We conclude that the rodent pinworm is an environmental agent that modifies the neonatal response to a self peptide, resulting in termination of the tolerance state and induction of a strong Th2-associated autoimmune disease and T cell memory.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/parasitología , Memoria Inmunológica , Receptores de Superficie Celular , Autotolerancia , Animales , Animales Recién Nacidos , Antinematodos/farmacología , Autoantígenos/inmunología , Enfermedades Autoinmunes/patología , Células Cultivadas , Citocinas/biosíntesis , Proteínas del Huevo/inmunología , Enterobiasis/complicaciones , Enterobiasis/tratamiento farmacológico , Eosinofilia/parasitología , Eosinofilia/patología , Femenino , Fenbendazol/farmacología , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos C57BL , Ooforitis/inmunología , Ooforitis/parasitología , Ooforitis/patología , Células Th2/inmunología , Células Th2/metabolismo , Uveítis/inmunología , Glicoproteínas de la Zona PelúcidaRESUMEN
Female (C57BL/6xA/J)F(1) mice undergoing thymectomy on day 3 after birth (d3tx) developed autoimmune ovarian disease (AOD) and autoimmune disease of the lacrimal gland. As both were prevented by normal adult CD25(+) T cells, regulatory T cell depletion is responsible for d3tx diseases. AOD began as oophoritis at 3 wk. By 4 wk, AOD progressed to ovarian atrophy with autoantibody response against multiple oocyte Ag of early ontogeny. The requirement for immunogenic endogenous ovarian Ag was investigated in d3tx female mice, d3tx male mice, and d3tx neonatally ovariectomized (OX) females. At 8 wk, all mice had comparable lacrimalitis but only those with endogenous ovaries developed AOD in ovarian grafts. The duration of Ag exposure required to initiate AOD was evaluated in d3tx mice OX at 2, 3, or 4 wk and engrafted with an ovary at 4, 5, or 6 wk, respectively. The mice OX at 2 wk did not have oophoritis whereas approximately 80% of mice OX at 3 or 4 wk had maximal AOD, thus Ag stimulus for 2.5 wk following d3tx is sufficient. AOD progression requires additional endogenous Ag stimulation from the ovarian graft. In mice OX at 3 wk, ovaries engrafted at 5 wk had more severe oophoritis than ovaries engrafted at 6 or 12 wk; moreover, only mice engrafted at 5 wk developed ovarian atrophy and oocyte autoantibodies. Similar results were obtained in mice OX at 4 wk. Thus endogenous tissue Ag are critical in autoimmune disease induction and progression that occur spontaneously upon regulatory T cell depletion.
Asunto(s)
Autoantígenos/fisiología , Enfermedades Autoinmunes/inmunología , Oocitos/inmunología , Ooforitis/inmunología , Complicaciones Posoperatorias/inmunología , Timectomía , Animales , Animales Recién Nacidos/inmunología , Atrofia , Autoanticuerpos/biosíntesis , Autoantígenos/administración & dosificación , Enfermedades Autoinmunes/etiología , Enfermedades Autoinmunes/patología , Femenino , Esquemas de Inmunización , Riñón , Masculino , Ratones , Ratones Endogámicos A , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones SCID , Oocitos/trasplante , Ooforitis/etiología , Ooforitis/patología , Ovario/patología , Complicaciones Posoperatorias/etiología , Complicaciones Posoperatorias/patología , Trasplante HeterotópicoRESUMEN
Research on murine autoimmune ovarian disease (AOD) models suggests that the following sequence of events operate in prevention and induction of AOD. Potentially pathogenic T cells for oocyte antigens that exist in normal mice are kept in check by regulatory CD25(+) T cells. Oocyte-specific pathogenic T cells are activated when the regulation is lost, as after day 3 thymectomy, or when T cells are stimulated through molecular mimicry. Activated, proinflammatory T cells induce interstitial ovarian inflammation without disruption in ovarian function. Activated T cells also help B cells that respond to endogenous oocyte antigens, to produce oocyte autoantibodies of diversified specificities. Autoantibodies, nonpathogenic in themselves, retarget T cell-mediated inflammation to ovarian follicles resulting in ovarian atrophy and ovarian failure. Future studies should determine the applicability of these findings to human ovarian autoimmunity.
Asunto(s)
Enfermedades Autoinmunes/prevención & control , Enfermedades del Ovario/inmunología , Enfermedades del Ovario/prevención & control , Receptores de Superficie Celular , Animales , Autoanticuerpos/inmunología , Autoantígenos/inmunología , Linfocitos T CD4-Positivos/inmunología , Proteínas del Huevo/inmunología , Femenino , Humanos , Tolerancia Inmunológica , Glicoproteínas de Membrana/inmunología , Linfocitos T/inmunología , Glicoproteínas de la Zona PelúcidaRESUMEN
In budding yeast, many mutants defective in meiotic recombination and chromosome synapsis undergo checkpoint-mediated arrest at the pachytene stage of meiotic prophase. We recovered the NDT80 gene in a screen for genes whose overexpression bypasses the pachytene checkpoint. Ndt80 is a meiosis-specific transcription factor that promotes expression of genes required for exit from pachytene and entry into meiosis I. Herein, we show that the Ndt80 protein accumulates and is extensively phosphorylated during meiosis in wild type but not in cells arrested at the pachytene checkpoint. Our results indicate that inhibition of Ndt80 activity is one mechanism used to achieve pachytene arrest.
Asunto(s)
Ciclo Celular , Proteínas de Unión al ADN , Proteínas Fúngicas/metabolismo , Meiosis , Proteínas de Saccharomyces cerevisiae , Factores de Transcripción/metabolismo , Mapeo Cromosómico , Cromosomas Fúngicos , Fosforilación , Plásmidos , Saccharomyces cerevisiae/citología , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismoRESUMEN
Estrogen receptors (ERs) are expressed in the thymus of both males and females, but their role in thymic development and function is unclear. To determine whether ERalpha plays a role in thymic function of either males or females, we compared thymuses of male and female wild-type (WT) and ERalpha knockout (alphaERKO) mice from birth to adulthood. Although thymic size was similar in both male and female WT and alphaERKO mice at birth (d 0), by postnatal d 5 and at all subsequent ages, both male and female alphaERKO mice had significant (30-55%) reductions in thymic weight. Morphometric analysis revealed a reduction in thymic medullary areas in adult alphaERKO mice compared with age-matched WT controls that paralleled thymic involution. There were changes in relative percentages of CD4+ and CD4+CD8+ T-cells, and large decreases (70-80%) in overall absolute numbers of CD4+ and CD4+CD8+ T-cells. Serum corticosterone and testosterone levels were not different in either neonatal or adult male WT or alphaERKO mice, and serum levels of 17beta-estradiol (E2) were similar in neonatal WT and alphaERKO males, indicating that increases in these thymolytic hormones are not responsible for the decreased thymic weight in alphaERKO males. Additionally, delayed-type hypersensitivity was significantly increased in male alphaERKO mice compared with WT mice. In summary, ERalpha deficiency does not inhibit initial differentiation or fetal thymic development, but the absence of ERalpha results in marked decreases in thymic size in both sexes during the postnatal period. These results are the first direct demonstration that the E2/ERalpha signaling system is necessary for maintenance of normal postnatal function of the female thymus gland. The similar results obtained in males demonstrate a role for the E2/ERalpha signaling system in the male thymus and emphasize that estrogens play a more critical role in the male than previously realized.
Asunto(s)
Estrógenos/fisiología , Receptores de Estrógenos/fisiología , Transducción de Señal , Timo/crecimiento & desarrollo , Animales , Animales Recién Nacidos/sangre , Linfocitos T CD4-Positivos , Linfocitos T CD8-positivos , Corticosterona/sangre , Estradiol/sangre , Receptor alfa de Estrógeno , Femenino , Hipersensibilidad Tardía , Recuento de Linfocitos , Masculino , Ratones , Ratones Noqueados , Receptores de Estrógenos/deficiencia , Receptores de Estrógenos/genética , Caracteres Sexuales , Testosterona/sangre , Timo/embriología , Timo/fisiologíaRESUMEN
To understand the pathogenesis of organ-specific autoimmune disease requires an appreciation of how the T cell-mediated inflammation is targeted, and how the organ function is compromised. In this study, autoantibody was documented to influence both of these parameters by modulating the distribution of T cell-mediated inflammation. The murine autoimmune ovarian disease is induced by immunization with the ZP3330-342 peptide of the ovarian zona pellucida 3 glycoprotein, ZP3. Passively transferred or actively induced Ab to ZP3335-342 bound to the zona pellucida in the functional and degenerative ovarian follicles, and the ovaries remained histologically normal. Transfer of ZP3330-342 peptide-specific T cells targeted the degenerative follicles and spared the functional follicles, and the resultant interstitial oophoritis was associated with unimpaired ovarian function. Unexpectedly, the coexistence of ZP3330-342 peptide-specific T cells and zona-bound autoantibody led to a dramatic translocation of the ovarian inflammation to the growing and mature ovarian follicles, with destruction of the ovarian functional unit. Ab retargeted both Th1-induced mononuclear inflammation and Th2-induced eosinophilic inflammation, and retargeting was induced by murine and rat polyclonal Abs to multiple distinct native B cell determinants of the zona pellucida. Therefore, by reacting with the native determinants in tissue Ag, Ab alters the distribution of T cell-mediated inflammation, and results in destruction of the functional units of the target organ. We propose that this is a clinically important and previously unappreciated element of Ab action in autoimmune disease.
Asunto(s)
Autoanticuerpos/fisiología , Ooforitis/inmunología , Ooforitis/patología , Receptores de Superficie Celular , Linfocitos T/inmunología , Linfocitos T/patología , Secuencia de Aminoácidos , Animales , Especificidad de Anticuerpos , Atrofia , Autoanticuerpos/administración & dosificación , Enfermedades Autoinmunes/etiología , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/patología , Citocinas/fisiología , Proteínas del Huevo/administración & dosificación , Proteínas del Huevo/inmunología , Epítopos de Linfocito T/inmunología , Femenino , Humanos , Inyecciones Subcutáneas , Transfusión de Linfocitos , Glicoproteínas de Membrana/administración & dosificación , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos A , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Ooforitis/etiología , Especificidad de Órganos/inmunología , Folículo Ovárico/inmunología , Folículo Ovárico/patología , Linfocitos T/trasplante , Zona Pelúcida/inmunología , Zona Pelúcida/patología , Glicoproteínas de la Zona PelúcidaRESUMEN
Endogenous Ag requirement for induction and maintenance of T cell tolerance has been extensively investigated in mice that express a transgenic Ag and/or its cognate transgenic TCR. In contrast, studies on tolerance for physiologically expressed self Ag and normal T cells are limited. Herein, we showed that the murine ovarian-specific ZP3 Ag is detectable from birth. Tolerance to ZP3 is detected in female relative to male mice. In comparison to males, 100-fold more ovarian peptide (pZP3) is required to elicit a comparable pathogenic response in females. Female tolerance to pZP3 was dependent on the presence of endogenous ovarian Ag, because neonatal ovariectomy converted the female response to that of males. Moreover, in female mice that were ovariectomized from the ages of 1-6 wk, the pZP3 responses were enhanced to the male level if ovaries were removed up to 7 days, but not 3 days, before adult challenge with pZP3. Thus, the physiologically expressed ZP3 Ag induces tolerance to pZP3, and the maintenance of tolerance is critically dependent on the continuous presence of the endogenous ovarian Ag. In contrast, exposure to endogenous ovarian Ag confined to the neonatal period is insufficient for the induction and maintenance of tolerance to ZP3.
Asunto(s)
Autoantígenos/fisiología , Proteínas del Huevo/inmunología , Glicoproteínas de Membrana/inmunología , Receptores de Superficie Celular , Autotolerancia/inmunología , Factores de Edad , Secuencia de Aminoácidos , Animales , Animales Recién Nacidos/crecimiento & desarrollo , Animales Recién Nacidos/inmunología , Animales Recién Nacidos/metabolismo , Autoantígenos/biosíntesis , Autoantígenos/metabolismo , Proteínas del Huevo/biosíntesis , Proteínas del Huevo/metabolismo , Proteínas del Huevo/fisiología , Femenino , Masculino , Glicoproteínas de Membrana/biosíntesis , Glicoproteínas de Membrana/metabolismo , Glicoproteínas de Membrana/fisiología , Ratones , Ratones Endogámicos A , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Ooforitis/inmunología , Ooforitis/metabolismo , Caracteres Sexuales , Factores de Tiempo , Zona Pelúcida/inmunología , Zona Pelúcida/metabolismo , Glicoproteínas de la Zona PelúcidaRESUMEN
CD9 is a tetraspan protein that associates with several beta1 integrins, including alpha6beta1. Because alpha6beta1 is present on murine eggs and interacts with the sperm-surface glycoprotein ADAM 2 (fertilin beta), we first asked whether CD9 is present on murine eggs and whether it functions in sperm-egg binding and fusion. CD9 is present on the plasma membrane of oocytes in the ovary as well as on eggs isolated from the oviduct. The anti-CD9 mAb, JF9, potently inhibits sperm-egg binding and fusion in vitro in a dose-dependent manner. JF9 also disrupts binding of fluorescent beads coated with native fertilin or a recombinant fertilin beta disintegrin domain. (Both ligands bind to the egg via alpha6beta1.) Immunohistochemistry showed that CD9 is undetectable in the uterine epithelium, appears basolaterally and as prominent apical patches on the epithelium in the region between the uterus and the oviduct, and then persists apically in the oviduct. The integrin alpha6A subunit is found in similar apical patches in the region between the uterus and oviduct, but is confined to the basal aspect of the epithelium in the uterus and oviduct. Hence, alpha6A and CD9 both are expressed on the apical epithelial surface at the uterine-oviduct junction. These findings correlate with the observation that fertilin beta "knockout" sperm traverse the uterus but do not progress into the oviduct, contributing to the infertility of fertilin beta(-/-) male mice. Our results suggest that high-avidity binding between fertilin beta (ADAM 2) and alpha6beta1 requires cooperation between alpha6beta1 and CD9. Such cooperation may assist sperm passage into the oviduct as well as sperm-egg interactions.
Asunto(s)
Antígenos CD/fisiología , Fertilización , Integrinas/metabolismo , Glicoproteínas de Membrana/metabolismo , Metaloendopeptidasas/metabolismo , Óvulo/metabolismo , Espermatozoides/metabolismo , Proteínas ADAM , Animales , Femenino , Fertilinas , Inmunohistoquímica , Integrina alfa6beta1 , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos ICR , Modelos Biológicos , Folículo Ovárico/metabolismo , Óvulo/citología , Pruebas de Precipitina , Unión Proteica , Tetraspanina 29RESUMEN
The detection of noninfectious ovarian inflammation (oophoritis) and serum ovarian autoantibodies in a patient with premature ovarian failure is indicative of an autoimmune etiology. The mechanisms of autoimmune ovarian injury leading to loss of function are currently unknown. In this study we investigated the impact of oophoritis on ovarian function based on two murine autoimmune ovarian disease (AOD) models. AOD can be induced by thymectomy at Day 3 after birth (d3tx). D3tx mice develop ovarian inflammation and atrophy with loss of oocytes. In these mice, ovarian atrophy and not oophoritis correlated with abnormal estrous cyclicity. The second AOD model is induced by active immunization of adult mice with a murine ZP3 peptide (pZP3) in adjuvant. After active immunization, the zona pellucida antibody titer, not oophoritis, correlated with reduced fertility. To investigate the effect of oophoritis in the absence of antibody response or ovarian atrophy, pZP3-specific T cells were passively transferred into naive syngeneic mice. This recruited cytokine-producing cells into the ovaries so that elevated cytokine production and its effect on ovarian function could be examined. Recipients of pZP3-specific T cells developed severe granulomatous oophoritis, and the diseased ovaries had elevated ovarian mRNA levels of interferon-gamma, interleukin-1beta, and tumor necrosis factor alpha. Despite these changes, fertility rates and gonadotropin-induced follicular development remained essentially normal. Therefore, normal ovarian function is compatible with severe ovarian inflammation mediated by autoreactive T cells.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Ooforitis/inmunología , Ovario/inmunología , Ovario/fisiopatología , Receptores de Superficie Celular , Células TH1/inmunología , Animales , Autoanticuerpos/sangre , Enfermedades Autoinmunes/fisiopatología , Citocinas/biosíntesis , Proteínas del Huevo/inmunología , Estro , Femenino , Infertilidad Femenina/inmunología , Interferón gamma/genética , Interleucina-1/genética , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos A , Ratones Endogámicos C57BL , Ooforitis/fisiopatología , ARN Mensajero/metabolismo , Timectomía , Factor de Necrosis Tumoral alfa/genética , Zona Pelúcida/inmunología , Glicoproteínas de la Zona PelúcidaRESUMEN
In this study, we have mapped and characterized a B cell epitope of sulfated glycoprotein ZP2 (ZP2) as a step toward the development of a multi-epitope zona pellucida (ZP) vaccine. Recombinant polypeptides expressed by random deoxyribonuclease-digested fragments of ZP2 cDNA were screened for binding to IE-3, a monoclonal antibody to murine ZP2. Positive clones contained cDNA inserts encoding polypeptide corresponding to ZP2(103-134). When normal or ovariectomized female mice were immunized with three overlapping peptides that span this region of ZP2 (101-120, 111-130, 121-140), only ZP2(121-140) elicited IgG antibodies that reacted with mouse ovarian ZP, indicative of the presence of native B epitope and helper T cell epitope in ZP2(121-140). To more finely map the ZP2 B cell epitope, a random peptide display library was screened with the IE-3 antibody, and a consensus tetramer sequence VxYK that matched the ZP2(123-126) sequence VRYK was located. Competitive immunofluorescence analysis with single alanine-substituted VxYK peptides ranked the relative contribution of the three critical B cell epitope residues as Y > V > K. A chimeric peptide was constructed that contained the YRYK motif of ZP2 and a bovine RNase T cell epitope. Although (C57BL/6xA/J) F1 (B6AF1) female mice immunized with the chimeric peptide developed ZP antibody response, this peptide elicited antibody only in mice of the histocompatibility complex (MHC) H-2(k or b) haplotype. In contrast, ZP2(121-140) peptide elicited antibody in inbred mice with three additional mouse MHC haplotypes. Moreover, although ZP2(121-140) contained a T cell epitope, no oophoritis was observed after immunization of B6AF1 mice with ZP2(121-140) in complete Freund's adjuvant (CFA). In a preliminary trial, female B6AF1 mice immunized with ZP2(121-140) in CFA had reduced litter sizes as compared with mice injected with CFA alone.
Asunto(s)
Anticoncepción Inmunológica , Proteínas del Huevo/inmunología , Glicoproteínas de Membrana/inmunología , Receptores de Superficie Celular , Vacunas , Secuencia de Aminoácidos , Animales , Proteínas del Huevo/química , Mapeo Epitopo , Epítopos/química , Epítopos/inmunología , Femenino , Inmunoglobulina G/inmunología , Masculino , Glicoproteínas de Membrana/química , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Datos de Secuencia Molecular , Ovariectomía , Proteínas Recombinantes de Fusión/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Zona Pelúcida/inmunología , Glicoproteínas de la Zona PelúcidaRESUMEN
Anti-Ro60 autoantibodies are found in a variety of autoimmune disorders including systemic lupus erythematosus (SLE), Sjögren's syndrome, primary biliary cirrhosis, and active hepatitis. They are the most prevalent autoantibodies in normal individuals and in asymptomatic mothers of infants afflicted with neonatal lupus. In the present study, immune responses to recombinant human Ro60 (rhRo60) and recombinant mouse Ro60 (rmRo60) and selected Ro60 peptides in non-SLE-prone mice were investigated. Multiple T and B cell epitopes were identified in Ro60. Immunizations with either xenogeneic or autologous Ro60 induced autoantibodies to a diverse group of autoantigens. In addition to La and Ro52, proteins in the small nuclear ribonucleoprotein (snRNP) particles such as SmA, SmB, SmD, and 70-kD U1-RNP were unexpectedly identified as targeted antigens. In the studies involving synthetic Ro60 peptides, both human and mouse Ro60316-335 peptides, which differ in three amino acids, were found to contain dominant cross-reactive T cell determinants. Immunizations with these peptides induced autoantibodies to Ro60, La, SmD, and 70-kD U1-RNP without autoantibodies to Ro52, SmA, or SmB. With human Ro60316-335 as the immunogen, additional autoantibodies reactive with the Golgi complex were found. In contrast to the immunodominance of both human and mouse Ro60316-335 peptides, the T cell determinant in human Ro60441-465 was dominant, whereas that in the mouse peptide was cryptic. Immunization with human Ro60441-465 induced primarily anti-peptide Abs. Mouse Ro60441-465 failed to induce an antibody response. These results show that both the nature of the immunogen and the immunogenicity of the related endogenous antigen are important in determining the specificities of the autoantibodies generated. They have significant implications for proposed mechanisms on the generation of complex patterns of autoantibodies to a diverse group of autoantigens in SLE patients.
Asunto(s)
Especificidad de Anticuerpos , Autoanticuerpos/inmunología , Autoantígenos/inmunología , Lupus Eritematoso Sistémico/inmunología , Fragmentos de Péptidos/inmunología , ARN Citoplasmático Pequeño , Ribonucleoproteínas/inmunología , Animales , Autoantígenos/genética , Linfocitos B/inmunología , Reacciones Cruzadas , Epítopos , Femenino , Aparato de Golgi/inmunología , Humanos , Epítopos Inmunodominantes , Ratones , Ratones Endogámicos BALB C , Fragmentos de Péptidos/genética , Proteínas Recombinantes/inmunología , Ribonucleoproteínas/genética , Ribonucleoproteínas Nucleares Pequeñas/inmunología , Especificidad de la Especie , Linfocitos T/inmunología , Vacunación , Antígeno SS-BRESUMEN
The yeast Zip1 protein (Zip1p) is a component of the central region of the synaptonemal complex (SC). Zip1p is predicted to form a dimer consisting of a coiled-coil domain flanked by globular domains. To analyze the organization of Zip1p within the SC, in-frame deletions of ZIP1 were constructed and analyzed. The results demonstrate that the C terminus but not the N terminus of Zip1p is required for its localization to chromosomes. Deletions in the carboxy half of the predicted coiled-coil region cause decreases in the width of the SC. Based on these results, a model for the organization of Zip1p within the SC is proposed. zip1 deletion mutations were also examined for their effects on sporulation, spore viability, crossing over, and crossover interference. The results demonstrate that the extent of synapsis is positively correlated with the levels of spore viability, crossing over, and crossover interference. In contrast, the role of Zip1p in synapsis is separable from its role in meiotic cell cycle progression. zip1 mutants display interval-specific effects on crossing over.
Asunto(s)
Cromosomas Fúngicos/genética , Proteínas Fúngicas/genética , Meiosis/genética , Mutagénesis Sitio-Dirigida , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Anticuerpos Antifúngicos , Cromosomas Fúngicos/química , Cromosomas Fúngicos/inmunología , Cromosomas Fúngicos/metabolismo , Intercambio Genético/genética , Proteínas Fúngicas/inmunología , Proteínas Fúngicas/metabolismo , Proteínas Nucleares , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Sistemas de Lectura/genética , Esporas Fúngicas/genética , Coloración y Etiquetado , Complejo Sinaptonémico/genéticaRESUMEN
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
Autoimmune diseases are often manifested as organ inflammation with loss of function, and detectable autoreactive T cell and autoantibody responses. In the proper genetic context, we have shown that these parameters of autoimmunity can result from a single pivotal event: the induction of a strong and persistent T cell response for a foreign or unrelated self peptide that mimics the target self peptide. This may apply to organ-specific and systemic autoimmunity, independent of whether the tissue inflammation results from T cell immune mechanism or antibodies. T cell peptide mimicry, through sharing of critical residues or by a less defined mechanism, can result in autoimmune disease. Once triggered, the helper T cell response leads rapidly to a concomitant autoantibody response spreading to distant B cell determinants of the self protein antigen. Evidently, with T cell help, endogenous antigens can stimulate B cells to provoke a functional autoantibody response against conformational antigenic determinants. These findings are based on recent studies on a novel autoimmune ovarian disease model induced by a self peptide with well-defined T and B cell epitopes. However, studies reported on systemic lupus erythematosus models have shown that similar events may result in autoantibody response in systemic autoimmunity.
Asunto(s)
Autoanticuerpos/biosíntesis , Enfermedades Autoinmunes/etiología , Ovario/inmunología , Receptores de Superficie Celular , Linfocitos T/fisiología , Animales , Proteínas del Huevo/inmunología , Epítopos de Linfocito T , Femenino , Humanos , Tolerancia Inmunológica , Activación de Linfocitos , Glicoproteínas de Membrana/inmunología , Ooforitis/etiología , Glicoproteínas de la Zona PelúcidaRESUMEN
Neonatal thymectomy (NTX) of BALB/c mice between days 1 and 3 post-birth leads to a high incidence of autoimmune gastritis involving T cells and autoantibodies. Although progress has been made in understanding various immunological events associated with that disease process, much remains to be learned about the regulatory factors which control autoimmune gastritis. In this study we have examined the potential for neuroendocrine hormones of the hypothalamus-pituitary-thyroid (HPT) axis to alter the outcome of experimental autoimmune gastritis in NTX mice. As reported here, thyroxine administered to young adult NTX mice during an active phase of disease development dramatically reduced the incidence of gastritis and the overall severity of disease, and lowered the levels of anti-parietal cell antibodies. In contrast, treatment of young NTX mice with thyroxine or other HPT hormones prior to the onset of disease had no beneficial effect on gastritis, but instead resulted in significantly higher anti-parietal cell antibody levels compared to non-hormone-treated NTX mice or to NTX mice treated as adults. Reverse transcriptase spectratype analyses of 22 Vbeta gene junctional sites revealed pronounced oligoclonality and limited junctional diversity in stomach lymphocytes from untreated NTX mice compared to normal mice or thyroxine-treated NTX mice. These findings identify a set of dynamic immune-endocrine interactions, linked to critical development-dependent events, that are involved in the expression and regulation of peripheral autoimmunity.