RESUMO
Objective: To explore the relationship between insomnia and osteoporosis. Methods: Mendelian randomization (MR) analysis were used in this study. The single nucleotide polymorphisms (SNPs) related to insomnia from genome-wide association analysis research data were selected as the instrumental variables by using inverse variance weighted (IVW), MR-Egger regression, weighted median method, maximum likelihood, penalized weighted median estimator, and Mendelian randomization robust adjusted profile score (MR-RAPS) to determine the causal relationship between insomnia and osteoporosis. Odds ratio (OR) and 95% confidence interval (CI) values were used to evaluate the association between insomnia and osteoporosis. Cochran's Q-test was used to detect heterogeneity of SNPs, MR-Egger regression was used to test for level pleiotropy, and the leave-one-out method was used to test sensitivity, MR pleiotropy residual sum and outlier (MR-PRESSO) method and radial MR were used to detect erroneous outliers. Results: The screening criteria were set based on the three major assumptions of MR; finally, 31 SNPs were included in the MR analysis. The results of MR causal effect analysis using the IVW method showed that insomnia increased the risk of osteoporosis by about 0.7% (OR=1.007, 95%CI 1.001-1.014, P=0.044); heterogeneity testing showed heterogeneity between SNPs (Q=57.91, P<0.001); and the MR- Egger intercept test did not indicate horizontal pleiotropy in this study (intercept value=3.807×10-5, P=0.888). Leave-one-out method showed that no single SNP had a significant impact on the overall results. No abnormal SNP was detected according to the MR-PRESSO results (P=0.059), and radial MR did not detect any outliers. Conclusion: Mendelian randomization analysis showed that insomnia can increase the risk of osteoporosis.
Assuntos
Estudo de Associação Genômica Ampla , Análise da Randomização Mendeliana , Osteoporose , Polimorfismo de Nucleotídeo Único , Distúrbios do Início e da Manutenção do Sono , Humanos , Osteoporose/genética , Osteoporose/epidemiologia , Distúrbios do Início e da Manutenção do Sono/genética , Distúrbios do Início e da Manutenção do Sono/epidemiologia , Fatores de RiscoRESUMO
Objective: To investigate the correlation between serum C-peptide and in adult population, and establish the corresponding insulin values of serum C-peptide levels. Methods: Cross-sectional study. The clinical data of the adults who underwent physical examination in the Second Medical Center of PLA General Hospital from January 2017 to December 2021 were retrospectively included. The participants were divided into type 2 diabetes group, prediabetes group and normal plasma glucose group according to the diagnostic criteria for diabetes. The correlation between serum C-peptide and insulin was explored by Pearson correlation analysis, linear regression analysis, and nonlinear regression analysis, and the corresponding insulin values of serum C-peptide were established. Results: A total of 48 008 adults were enrolled, including 31 633 males (65.9%) and 16 375 females (34.1%), aged (50.1±9.9) years (18-89 years). There were 8 160 subjects (17.0%) with type 2 diabetes, 13 263 subjects (27.6%) with prediabetes, and 26 585 subjects (55.4%) with normal plasma glucose. The serum fasting C-peptide (FCP, M(Q1, Q3)] of the three groups were 2.76(2.18, 3.47), 2.54(1.99, 3.21) and 2.18(1.71, 2.79)µg/L, respectively. The fasting insulin [FINS, M(Q1,Q3)] of the three groups were 10.98(7.57, 16.09), 10.06(6.95, 14.47) and 8.43(5.86,12.12)mU/L, respectively. FCP was positively correlated with FINS (r=0.82), and 2 h postprandial C-peptide (2 h CP) was positively correlated with 2 h postprandial insulin (2 h INS) (r=0.84) (both P<0.001). FCP was linearly associated with FINS (R2=0.68), and 2 h CP was linearly associated with 2 h INS (R2=0.71) (both P<0.001). There was a power function correlation between FCP and FINS (R2=0.74), and 2 h CP and 2 h INS (R2=0.78) (both P<0.001). The results of the statistical analysis were similar in various glucose metabolism subgroups. Since the fitting degree of the power function model was higher than that of the linear model, the power function model was the best model. The power function equation was FINS=2.96×FCP1.32, and 2 h INS=1.64×(2 h CP)1.60, respectively. Multivariate linear regression analysis demonstrated that FCP was a related factor of FINS (R2=0.70, P<0.001) and 2 h CP was a related factor of 2 h INS (R2=0.73, P<0.001), after adjusting for related confounders. Conclusions: There was a power function correlation between FCP and FINS, 2 h CP and 2 h INS in adult population. The insulin values corresponding to C-peptide levels were established in the study.
Assuntos
Diabetes Mellitus Tipo 2 , Resistência à Insulina , Estado Pré-Diabético , Masculino , Feminino , Adulto , Humanos , Insulina/metabolismo , Peptídeo C , Glicemia/análise , Estudos Retrospectivos , Estudos TransversaisRESUMO
Objective: To investigate the relationship between hemoglobin and serum uric acid in adults with various glucose metabolism status. Methods: The demographic data and biochemical indicators of the adult population who had received physical examination in the Second Medical Center of the PLA General Hospital from January 2018 to December 2021 were collected. The subjects were divided into two groups according to the level of serum uric acid: the normal uric acid group and the hyperuricemia group. The relationship between hemoglobin (stratified into four levels of Q1 to Q4 by the quartile) and serum uric acid was quantified by using Pearson correlation and logistic regression analysis. The effects of age and glucose metabolism status on the relationship between hemoglobin and serum uric acid were analyzed. Results: A total of 33 183 adults were enrolled with age (50.6±10.0) years. The level of hemoglobin in the normal uric acid group (142.61±14.24) g/L was significantly lower than that in the hyperuricemia group [(151.79±11.24) g/L, P<0.001]. Univariate Pearson correlation analysis showed that hemoglobin was positively associated with serum uric acid (r=0.444, P<0.001). After adjusting for related confounding factors, multivariate logistic regression analysis showed that hemoglobin was associated with serum uric acid, and the OR values (95%CI) of hemoglobin Q2 to Q4 group were 1.29 (1.13-1.48), 1.42 (1.24-1.62) and 1.51 (1.32-1.72), respectively (Ptrend<0.001) when compared with hemoglobin Q1 group. Subgroup analysis and hierarchical interaction analysis suggested that with the increase of hemoglobin, the serum uric acid in the age<60 years subgroup, normal glucose subgroup and prediabetes subgroup increased gradually (Ptrend<0.05 and Pinteraction<0.001). Conclusion: The association between hemoglobin and serum uric acid in adults is affected by age and glucose metabolism status.
Assuntos
Hiperuricemia , Estado Pré-Diabético , Humanos , Adulto , Pessoa de Meia-Idade , Ácido Úrico , Hiperuricemia/epidemiologia , Hemoglobinas , Glucose , Fatores de RiscoRESUMO
OBJECTIVE: The aim of this study was to examine the effects of stent-assisted coiling embolization of middle cerebral artery (MCA) trifurcation wide-necked aneurysms in clinical practice. PATIENTS AND METHODS: 57 patients with MCA trifurcation wide-necked aneurysms underwent stent-assisted coiling embolization using a solitaire AB stent. All 57 patients completed the surgery successfully. Embolization efficacy was graded according to the modified Raymond scale. RESULTS: There were 52 cases of complete embolization, 4 cases of residual aneurysm neck, and 1 case of residual aneurysm body. 50 patients participated in a 6-36-month follow-up. There has not been observed any aneurysm rupture and hemorrhage. 50 patients received digital subtraction angiography (DSA) re-examination; 46 patients presenting complete embolization had no aneurysm relapses; 3 patients had residual aneurysm neck demonstrated; 1 patient had no aneurysm neck and others 2 were in stable condition. Finally, the patient with residual aneurysm body showed no sign during follow-up reexamination. CONCLUSIONS: Stent-assisted coiling embolization of intracranial wide-necked aneurysms using the solitaire AB stent was safe and effective.
Assuntos
Angiografia Digital/métodos , Embolização Terapêutica/métodos , Stents , Adulto , Idoso , Angiografia Cerebral/métodos , Feminino , Humanos , Aneurisma Intracraniano/cirurgia , Masculino , Pessoa de Meia-Idade , Artéria Cerebral Média , Resultado do TratamentoRESUMO
OBJECTIVE: This study aimed to investigate the expression and clinical significances of long noncoding RNA-ATB (lncRNA-ATB) in papillary thyroid cancer (PTC), and to explore the roles of lncRNA-ATB in PTC cell proliferation and migration. PATIENTS AND METHODS: The expression of lncRNA-ATB in 64 PTC tissues and paired adjacent noncancerous thyroid tissues was measured. The association between lncRNA-ATB expression and clinicopathological characteristics was analyzed by Pearson X2. The diagnostic value of lncRNA-ATB was evaluated by receiver operating characteristic curve (ROC) analyses. The effects of lncRNA-ATB on PTC cell proliferation were evaluated by Cell Counting Kit-8 assays and Ethynyl deoxyuridine incorporation assays. The effects of lncRNA-ATB on PTC cell migration were evaluated by transwell assays. RESULTS: LncRNA-ATB is upregulated in PTC tissues compared with noncancerous tissues. LncRNA-ATB is also increased in PTC cell lines compared with normal thyroid follicular epithelial cell line. High-expression of lncRNA-ATB is associated with large tumor size and lymph node metastasis. ROC analyses revealed that lncRNA-ATB could sensitively discriminate PTCs from noncancerous tissues, as well as discriminating PTCs with lymph node metastasis from those without lymph node metastasis. Functional experiments showed that depletion of lncRNA-ATB significantly inhibits PTC cell proliferation and migration. CONCLUSIONS: LncRNA-ATB is upregulated and functions as an oncogene in PTC. Furthermore, lncRNA-ATB may serve as a diagnostic biomarker and therapeutic target for PTC.
Assuntos
Carcinoma Papilar/patologia , RNA Longo não Codificante/fisiologia , Neoplasias da Glândula Tireoide/patologia , Adulto , Idoso , Carcinoma Papilar/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Feminino , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , RNA Longo não Codificante/análise , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide/genéticaRESUMO
Objective: To discuss functional connectivity changes in the emotional network of patients with post-concussion syndrome (PCS) and their clinical significance by resting-state functional magnetic resonance imaging (rs-fMRI). Methods: Twenty-seven patients with PCS were recruited from the Department of Neurosurgery of Anhui provincial hospital affiliated to Anhui medical university from October 2015 to April 2016, and 27 healthy subjects were recruited as the controls. The Hamilton Anxiety Scale (HAMA) and The Hamilton Depression Scale (HAMD) were used to evaluate the emotional state of two groups of subjects. All fMRI data were preprocessed after RS-fMRI scanning, the left and right amygdala were selected as region of interest (ROI) to make functional connectivity (FC) calculation with the whole brain and then the results were did statistical analysis in order to obtain the altered brain areas of amygdala and whole brain functional connectivity in the PCS patient, to understand the functional changes of emotional network. Results: HAMA and HAMD scores of PCS group and the health controls had significant statistical difference (HAMA: the PCS group 9.8±1.5, the health controls 4.5±1.2, P=0.044; HAMD: the PCS group 12±1.2, the health controls was 4.2±1.5, P=0.024). Compared with the health controls, the left amygdala in PCS patients showed decreased FC with left insula, left putamen, left anterior cingulate gyrus, left inferior orbital frontal gyrus, left medial superior frontal gyrus, bilateral superior temporal gyrus, left superior temporal pole, bilateral supramarginal gyrus et al, on the contrary with the increased FC with right superior orbital frontal gyrus, right middle frontal lobe, right orbital frontal lobe, right middle frontal gyrus. The right amygdala in PCS patients showed decreased FC with bilateral putamen, right inferior orbital frontal gyrus, left insula, bilateral precuneus, bilateral superior temporal pole, right superior temporal gyrus, right supramarginal gyrus et al. Similarly, it had the increased FC with the left middle occipital lobe and the left superior occipital lobe. Conclusion: Abnormal emotional network function of PCS patients in resting state, which may be one of the reasons that lead to emotional and cognitive dysfunction in PCS patients.
Assuntos
Mapeamento Encefálico , Emoções , Imageamento por Ressonância Magnética , Síndrome Pós-Concussão/fisiopatologia , Encéfalo , Estudos de Casos e Controles , Humanos , Síndrome Pós-Concussão/psicologiaRESUMO
OBJECTIVE: To aim at the efficacies of surgical clipping and endovascular embolization for oculomotor nerve palsy (ONP) as treatments for posterior communicating artery aneurysm (PcoAA), and the comparison and various influencing factors of the treatments. PATIENTS AND METHODS: An analysis of the clinical data of 52 enrolled PcoAA patients with ONP who had treatment in the Department of Neurosurgery in Anhui Provincial Hospital from January 2011 to June 2015 was conducted. There were 23 patients among a total underwent surgical clippings and others 29 patients received endovascular embolization treatment. Then, the age, gender, aneurysm size and rupture status, onset duration, preoperative ONP severity and postoperative recovery degree of ONP of patients in the two groups were compared. RESULTS: The final ONP outcomes of the 52 PcoAA patients consisted of 27 full recovery patients (51.9%), 21 partial recovery patients (40.4%), and 4 no recovery patients (7.7%). (1) Within the 23 patients in the surgical clipping group, subarachnoid hemorrhage (SAH) occurred in 16 patients, and no SAH occurrence in the other 7 patients; the final ONP evaluation showed 18 patients fully recovered (78.3%) and 5 patients partially recovered (21.7%). Within the 29 patients in the endovascular embolization group, SAH occurred in 18 patients, and no SAH occurrence in the other 11 patients; the final ONP evaluation showed 9 patients fully recovered (31%), 16 patients partially recovered in 16 patients (55.2%) and 4 no recovery patients (13.8%). (2) The postoperative ONP recovery was analyzed with multivariate logistic regression, and the treatment method was an independent factor for ONP recovery (OR = 0.041, 95% CI: 0.007-0.261, p < 0.01). CONCLUSIONS: When compared with the endovascular embolization, the surgical clipping showed a better efficacy in the recovery from PcoAA related ONP.
Assuntos
Embolização Terapêutica , Aneurisma Intracraniano/cirurgia , Doenças do Nervo Oculomotor/cirurgia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Paralisia/fisiopatologia , Recuperação de Função Fisiológica , Estudos Retrospectivos , Hemorragia Subaracnóidea , Resultado do TratamentoRESUMO
Hemangioblastoma of the central nervous system occurs as sporadic tumors or as a part of von Hippel-Lindau (VHL) disease, an autosomal dominant hereditary tumor syndrome caused by a germline mutation in the VHL tumor suppressor gene. We screened a Chinese family with VHL for mutations in the VHL gene and evaluated a genetic test for diagnosing VHL disease and clinical screening of family members. DNA extracted from the peripheral blood of all live members and from tissue of deceased family members with VHL disease was amplified by polymerase chain reaction to 3 VHL gene exons. Mutations in the amplification products were compared against the Human Gene Mutation Database. The involvement of multiple organs among the kindred with VHL disease was confirmed by medical history and radiography. Of the 12 members of the 4-generation family, 5 were diagnosed with VHL disease. Patient age at the initial diagnosis was 26-36 years (mean = 31 years). The mean time was 15 (11-19 months) from symptom appearance to the first patient visit to the hospital. Sequence analysis revealed that the frameshift mutation 327del C (p.Gly39Alafs*26) in exon 1 affected all family members, but not the healthy individuals or 16 unrelated controls. Members without gene mutation showed no clinical manifestation of VHL disease. We detected a conserved novel frameshift mutation in the VHL gene of the family members that contributes to VHL. DNA analysis of VHL is advantageous for VHL diagnosis. We developed a quick and reliable method for VHL diagnosis.
Assuntos
Mutação da Fase de Leitura , Hemangioblastoma/genética , Proteína Supressora de Tumor Von Hippel-Lindau/genética , Doença de von Hippel-Lindau/genética , Adulto , Análise Mutacional de DNA , Feminino , Testes Genéticos , Hemangioblastoma/diagnóstico , Hemangioblastoma/etiologia , Humanos , Masculino , Linhagem , Doença de von Hippel-Lindau/complicações , Doença de von Hippel-Lindau/diagnósticoRESUMO
The differential screening method was used to isolate the soy photoperiodic response-related genes and to further elucidate the molecular mechanisms of the soybean photoperiodic response. The light-sensitive species Zhong Dou 24 was used to receive long-time sunshine, short-time sunshine, and natural sunshine treatment. The cDNA-amplified fragment length polymorphism technique was used to screen the differentially expressed cDNA fragments. The rapid amplification of cDNA end technique was used to isolate the gene. Semi-quantitative reverse transcription polymerase chain reaction analysis was used to analyze the gene expression patterns in different light cycles. The gene had a total length of 983 bp, contained a complete open reading frame that encoded 248 amino acids, and shared homology with the mitochondrial phosphate transporter protein. The expression pattern analysis results showed that this gene was expressed in the early stages of soybean growth and development. The short-time sunshine inhibited its expression, whereas the long-time sunshine enhanced its expression. The differential screening method was used to isolate the soybean mitochondrial phosphate transporter gene. The gene may be used as a negative regulatory factor that is involved in the photoperiodic response of soybean.
Assuntos
Glycine max/genética , Proteínas de Transporte de Fosfato/genética , Proteínas de Plantas/genética , Clonagem Molecular , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Proteínas de Transporte de Fosfato/isolamento & purificação , Proteínas de Transporte de Fosfato/metabolismo , Fotoperíodo , Filogenia , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Análise de Sequência de DNA , Homologia de Sequência do Ácido NucleicoRESUMO
To investigate the relationship between the CYP1B1 L432V polymorphism, ERalpha and ERbeta positivities and the incidence of endometrial cancer. The relationship between CYP1B1 L432V polymorphism, ERalpha and ERbeta positivities and endometrial cancer was investigated using the allele-specific polymerase chain reaction method to analyze gene polymorphism in exon 3 codon 432 (C-G) of CYP1B1. Our results are as follows: in endometrial cancer cases the prevalence rates of CYP1B1 L432V genotypes C/C, C/G, and G/G were 47.2%, 36.1%, and 16.7%, respectively, and 68.8%, 23.8% and 7.5% in the control group, respectively. The frequencies of CYP1B1 C and G alleles were 65.3% and 34.7% in endometrial cancer patients and 80.6% and 19.4% in the control group. A significant difference was found in the genotype distributions or allele frequencies of CYP1B1 L432V polymorphism between the two groups (p < 0.05). Compared with wild-type C/C, the susceptibility of endometrial cancer with homozygotic mutation G/G and heterozygotic mutation C/G increased by 3.235 (95%CI 1.111-9.425) and 2.214 (95% CI 1.067-4.593). Moveover, the positive expression of ERalpha in genotypes G/G and C/G was higher than in the wild genotype C/C (p < 0.05). In conclusion, allelic polymorphism of CYP1B1 L432V increases the risk of endometrial cancer and has a positive correlation with ERalpha expression.
Assuntos
Adenocarcinoma de Células Claras/genética , Hidrocarboneto de Aril Hidroxilases/genética , Carcinoma Endometrioide/genética , Neoplasias do Endométrio/genética , Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/genética , Adenocarcinoma de Células Claras/metabolismo , Adulto , Idoso , Alelos , Hidrocarboneto de Aril Hidroxilases/metabolismo , Carcinoma Endometrioide/metabolismo , Citocromo P-450 CYP1B1 , Neoplasias do Endométrio/metabolismo , Receptor alfa de Estrogênio/metabolismo , Receptor beta de Estrogênio/metabolismo , Feminino , Frequência do Gene , Genótipo , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Razão de Chances , Polimorfismo Genético , RiscoRESUMO
Eighty-three subjects (47 women and 36 men) were submitted to Plutchik-van Praag's (PVP) depression inventory, Zuckerman's sensation seeking scales and Zuckerman-Kuhlman's personality inventory, and underwent auditory evoked potential studies using clicks at 4 different intensities of 70, 80, 90 and 100 dB. The clicks were delivered at an interstimulus interval varying randomly around 0.5s, which can elicit an obligatory subcomponent of N1. The P2 latency was significantly prolonged at the highest intensity. The intensity dependence of peak-to-peak N1-P2 and of baseline-to-peak N1 and P2 components was pronounced and the majority of subjects were augmenters. The N1 latency elicited at 70 dB was positively correlated with the thrill and adventure seeking, which then correlated the activity. The correlation suggests that a lower level of arousal, as indicated by prolonged N1 latency, would lead one to seek higher stimulation, such as the augmented response, the increased desire of physical thrill and adventure and elevated activity. This study, therefore, supports Zuckerman's theory that a sensation seeking personality is related to cortical arousal level.
Assuntos
Córtex Cerebral/fisiologia , Potenciais Evocados Auditivos/fisiologia , Testes Neuropsicológicos , Testes de Personalidade , Personalidade/fisiologia , Estimulação Acústica , Adulto , Eletroencefalografia , Eletroculografia , Feminino , Humanos , Masculino , Inventário de Personalidade , Escalas de Graduação Psiquiátrica , Tempo de Reação/fisiologia , Inquéritos e QuestionáriosRESUMO
Migraine sufferers have abnormal cerebral information processing and personality disorders, post-traumatic headache sufferers also have some personality changes. We therefore, studied intensity dependence of auditory evoked potentials, Plutchik-van Praag's depression inventory, Zuckerman's sensation seeking scales and Zuckerman-Kuhlman's personality questionnaire in patients suffering from migraine without aura (n = 26) and chronic post-traumatic headaches (n = 26) as well as in healthy volunteers (n = 30). The migraine group showed significantly increased neuroticism-anxiety than controls, increased intensity dependence of N1-P2, and decreased thrill and adventure score compared with the controls and post-traumatic headaches. The post-traumatic headache had significantly increased depression compared with the controls, and increased disinhibition compared with the controls and migraines. This study demonstrates that the two headache types have different neurophysiological and personality traits. The pronounced intensity dependence of N1-P2 suggests a cortical potentiation response, together with a decreased thrill and adventure seeking, favor a lower serotonergic innervation in migraine. While the elevated disinhibition and depression, as consequences, may be linked with the wide cortical neuronal/axonal degeneration in post-traumatic headache.
Assuntos
Traumatismos Craniocerebrais/complicações , Potenciais Evocados Auditivos , Cefaleia/fisiopatologia , Cefaleia/psicologia , Transtornos de Enxaqueca/fisiopatologia , Transtornos de Enxaqueca/psicologia , Adulto , Eletroencefalografia , Feminino , Cefaleia/etiologia , Humanos , Masculino , Testes Neuropsicológicos , Testes de Personalidade , Inquéritos e QuestionáriosAssuntos
Anticorpos Monoclonais/farmacologia , Antígenos de Superfície/fisiologia , Sobrevivência de Enxerto/fisiologia , Intestino Delgado/transplante , Veia Porta/imunologia , Transplante Homólogo/imunologia , Animais , Antígenos CD , Antígenos de Superfície/imunologia , Citocinas/análise , Citocinas/biossíntese , Camundongos , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos LewRESUMO
BACKGROUND: We studied the mechanism behind increased renal allotransplant survival when C3H mice received donor-specific portal vein or oral immunization with C57BL/6 cells. Both regimens lead to donor-specific increased graft survival, in association with decreased production of cytotoxic T lymphocytes and altered cytokine production from host lymphocytes (decreased interleukin [IL]-2 production; increased IL-4, IL-10, and transforming growth factor-beta). METHODS: We examined a role for persistent donor-derived antigen, in association with host dendritic cells, as well as a role for intercellular adhesion molecule-1 (ICAM-1), in the maintenance of unresponsiveness in host C3H spleen cells to donor antigen. We investigated whether there was a cooperative interaction between donor dendritic cells (DC) and host hepatic mononuclear cells in the induction of immunoregulation in C3H cells. RESULTS: In mice with surviving renal grafts, donor antigen, in association with host DC, induced the recall of cytotoxicity from C57BL/6 immune C3H spleen cells and IL-4 but not IL-2 production, despite the decreased cytotoxicity seen in the renal transplant recipients themselves. Fresh donor DC induced IL-2 but not IL-4 production. Blocking expression of ICAM-1 on donor grafts, either with anti-ICAM-1 monoclonal antibodies after renal grafting or using grafts from ICAM-1 "knockout" mice, led to further increased survival. Cultured C3H responder spleen cells, incubated with C57BL/6 DC and C3H hepatic cells, transferred hyporesponsiveness to C57BL/6 cells in vitro and in vivo (as assayed by survival of C57BL/6 renal allografts). CONCLUSIONS: Our data suggest a role for ICAM-1, persistent donor antigen (on host DC), and accessory hepatic monocytes in the induction and maintenance of tolerance after portal vein immunization.
Assuntos
Apresentação de Antígeno/imunologia , Sobrevivência de Enxerto/imunologia , Imunização Passiva , Molécula 1 de Adesão Intercelular/fisiologia , Isoantígenos/imunologia , Transplante de Rim/imunologia , Administração Oral , Animais , Células Dendríticas/imunologia , Feminino , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/prevenção & controle , Tolerância Imunológica/imunologia , Interleucina-10/sangue , Interleucina-2/sangue , Interleucina-4/sangue , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Camundongos Knockout , Veia Porta , Linfócitos T Citotóxicos/imunologia , Fator de Crescimento Transformador beta/sangueRESUMO
Female Lewis (LEW) rats received orthotopic small intestinal transplantation (SIT), or tail skin grafts from female (Lewis x Brown Norway)F1 (LBNF1) rats, along with peritransplant portal venous (pv) infusion of LBNF1 bone marrow-derived dendritic cells derived from male donors. All animals received im injection with cyclosporin A (5 mg/kg) for 3 consecutive days following transplantation. In some cases rats received intravenous injections, at 2-day intervals, with 1 mg of monoclonal antibodies to ICAM-1 or the integrins alpha 4, alpha L, or beta 2, or combinations of these reagents. Cells were harvested from the recipient rats at different times posttransplantation, and single cell suspensions were analyzed by FACS for expression of CD3+, CD4+, CD8+, alpha beta TcR+, and gamma delta TcR+ cells. Other tissue samples were used for histopathological assessment of rejection. We also investigated donor-specific and third-party (Wistar-Furth, Wi) restimulation of host lymphocytes from MLN, PLN, and PP for production of different cytokines in vitro. Of the various antibodies tested, only anti-alpha 4, but not anti-alpha L, -beta 2, nor -ICAM-1 led to further increased graft survival of LBNF1 SIT beyond that seen with pv-infused cells alone (30 days vs 19 days), while the combination of anti-alpha L (or beta 2) and ICAM-1 produced further significantly increased survival of skin grafts (30 days vs 21 days). For both SIT and skin-grafted animals increased graft survival was associated with decreased production of IL-2 and IFN-gamma and increased production of IL-4 and IL-10 from tissues local to the graft (PP and draining LN, respectively), with less significant alterations in tissues distant to the graft (PLN for SIT, and MLN for skin grafts). While, as reported previously, pv-immunized SIT rats showed increased gamma delta TCR+ cells within the SIT in association with increased graft survival, treatment with anti-alpha 4 diminished this increase in gamma delta TCR+ cells, while simultaneously increasing SIT survival. Nevertheless, the bias toward increased IL-10 production, and decreased IFN-gamma production, from cells of animals showing increased survival was maintained. These data suggest that local graft infiltration with gamma delta TCR+ cells following pv immunization is not necessary for prolongation of survival in this model system, although functional changes in the local cytokines milieu may be important.
Assuntos
Antígenos CD/fisiologia , Antígenos CD18/fisiologia , Rejeição de Enxerto , Molécula 1 de Adesão Intercelular/fisiologia , Intestino Delgado/transplante , Antígeno-1 Associado à Função Linfocitária/fisiologia , Transplante de Pele/imunologia , Animais , Anticorpos Monoclonais/imunologia , Citocinas/biossíntese , Feminino , Integrina alfa4 , Masculino , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos Lew , Ratos Wistar , Receptores de Antígenos de Linfócitos T gama-delta/fisiologia , Transplante Homólogo , Molécula 1 de Adesão de Célula Vascular/fisiologiaRESUMO
BACKGROUND: Portal venous (p.v.) peritransplant immunization with dendritic cells from bone marrow cultures, along with cyclosporine (10 mg/kg), produces antigen-specific increased renal allograft survival compared with recipients receiving intravenous (i.v.) immunization. Increased survival is associated with altered cytokine production from recipient T cells restimulated with donor antigen. We used a suppressive subtractive hybridization approach to explore a role in the regulation of transplant rejection for other genes differentially expressed after p.v. immunization. METHODS: Subtractive hybridization was performed using tissue from p.v. and i.v. immunized mice and a novel polymerase chain reaction-based approach. A gene-bank search was used to identify the source of the differentially expressed cDNAs. One product, the mouse homologue of rat OX-2, was further analyzed using Western gels and FACS analysis of dendritic cells (NLDC145+) isolated from p.v.-immunized mice. RESULTS: Eighty cDNA clones were obtained by suppressive subtractive hybridization. Differential expression was confirmed in Northern RNA blots. One clone showed sequence homology to a gene encoding a molecule on rat dendritic cells (MRC OX-2), with homology to genes encoding the costimulatory molecules CD80 (B7-1) and CD86 (B7-2). In p.v.-immunized mice, a monoclonal antibody to the rat OX-2 molecule identified, by Western blot analysis, increased expression of a molecule with molecular weight (43 kDa) analogous to rat MRC-OX-2; labels (by FACS analysis) indentified increased numbers of a population of cells staining with NLDC145; and blocks indentified increased graft survival. CONCLUSION: Our data suggest that OX-2 is functionally important in the increased graft survival seen in p.v.-immunized mice receiving renal allografts.
Assuntos
Antígenos de Superfície/biossíntese , Antígenos de Superfície/genética , Células Dendríticas/imunologia , Células Dendríticas/transplante , Sobrevivência de Enxerto/imunologia , Transplante de Rim/imunologia , Veia Porta/imunologia , Animais , Anticorpos Monoclonais , Antígenos CD , Antígenos de Superfície/análise , Sequência de Bases , Células da Medula Óssea/citologia , Células da Medula Óssea/imunologia , Clonagem Molecular , Citocinas/biossíntese , Primers do DNA , DNA Complementar , Humanos , Terapia de Imunossupressão/métodos , Hibridização In Situ , Isoantígenos/imunologia , Glicoproteínas de Membrana/biossíntese , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Ratos , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Linfócitos T/imunologia , Fatores de Tempo , Transplante HomólogoRESUMO
gamma delta TCR+ hybridoma cells prepared from mesenteric lymph node cells of animals receiving donor-specific immunization via the portal vein can adoptively transfer this increased graft survival to naive animals. Analysis of TCR gamma-chain junctional sequence diversity suggested that some 40 to 50% of the hybridomas expressed gamma-chain junctional sequence diversity and were stimulated to produce cytokines both by heat shock proteins and by minor histocompatibility Ag-specific irradiated peritoneal cells. The remaining gamma delta TCR+ hybridoma cells expressed TCR with a common gamma-chain junctional sequence and were stimulated to cytokine production by MHC-matched, but minor histocompatibility Ag-mismatched (as well as matched), peritoneal cells, but not by heat shock proteins. We have compared the effectiveness of representative hybridomas expressing unique gamma-chain junctional sequences or common gamma-chain junctional sequences for prolongation of donor-specific or third-party (MHC-matched or MHC-mismatched) skin grafts. Our data show a good correlation between the specificity for stimulation for cytokine production in vitro and efficacy in graft prolongation assays in vivo. Hybridoma cells expressing unique gamma-chain junctional sequences that showed Ag-specific stimulation of cytokine production in vitro and skin graft survival in vivo augmented survival of third-party skin grafts if simultaneously transplanted with both Ag-specific and third-party skin grafts. Graft prolongation in vivo using cells from either population of gamma delta TCR+ hybridomas was decreased by infusion of anti-IL-10 mAb and abolished when both anti-IL-10 and anti-TGF-beta Abs were used together.
Assuntos
Transferência Adotiva , Epitopos de Linfócito T/imunologia , Facilitação Imunológica de Enxerto , Hibridomas/metabolismo , Transfusão de Linfócitos , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Transplante de Pele , Doadores de Tecidos , Transferência Adotiva/métodos , Animais , Sequência de Bases , Citocinas/biossíntese , Epitopos de Linfócito T/biossíntese , Facilitação Imunológica de Enxerto/métodos , Hibridomas/transplante , Esquemas de Imunização , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Veia Porta , Receptores de Antígenos de Linfócitos T gama-delta/biossíntese , Transplante de Pele/métodosAssuntos
Rejeição de Enxerto/prevenção & controle , Intestino Delgado/transplante , Transfusão de Linfócitos , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Linfócitos T/imunologia , Transplante Homólogo/imunologia , Transferência Adotiva , Animais , Separação Celular , Rejeição de Enxerto/imunologia , Terapia de Imunossupressão/métodos , Ratos , Ratos Endogâmicos LewRESUMO
Portal venous (pv) transfusion before transplant with large numbers (100 x 10(6)) of irradiated multiple minor histoincompatible spleen cells (B10.Br) augments allogeneic skin graft survival in C3H mice. We have shown in earlier studies that this is correlated with preferential activation for production of type 2 cytokines (interleukin [IL]-4 and IL-10) and decreased production of type 1 cytokines (IL-2 and interferon [IFN] gamma). We have also shown that recombinant (r)IL-12, in association with anti-IL-10 monoclonal antibody, can reverse in vivo the graft prolongation afforded by pv immunization and the altered cytokine production that follows. Adoptive transfer of inhibition of graft rejection is possible at early times after pv immunization, using plastic adherent cells obtained from the liver of treated mice. We show below that within 4 days of pv immunization, dendritic cells (NLDC-145+) isolated from the thymus, mesenteric lymph node (MLN), and.