RESUMEN
In November 2022, 68% of the population received at least one dose of COVID-19 vaccines. Owing to the ongoing mutations, especially for the variants of concern (VOCs), it is important to monitor the humoral immune responses after different vaccination strategies. In this study, we developed a SARS-CoV-2 variant protein microarray that contained the spike proteins from the VOCs, e.g., alpha, beta, gamma, delta, and omicron, to quantify the binding antibody and surrogate neutralizing antibody. Plasmas were collected after two doses of matching AZD1222 (AZx2), two doses of matching mRNA-1273 (Mx2), or mixing AZD1222 and mRNA-1273 (AZ+M). The results showed a significant decrease of surrogate neutralizing antibodies against the receptor-binding domain in all VOCs in AZx2 and Mx2 but not AZ+M. A similar but minor reduction pattern of surrogate neutralizing antibodies against the extracellular domain was observed. While Mx2 exhibited a higher surrogate neutralizing level against all VOCs compared with AZx2, AZ+M showed an even higher surrogate neutralizing level in gamma and omicron compared with Mx2. It is worth noting that the binding antibody displayed a low correlation to the surrogate neutralizing antibody (R-square 0.130-0.382). This study delivers insights into humoral immunities, SARS-CoV-2 mutations, and mixing and matching vaccine strategies, which may provide a more effective vaccine strategy especially in preventing omicron.
Asunto(s)
Vacunas contra la COVID-19 , COVID-19 , Humanos , SARS-CoV-2 , ChAdOx1 nCoV-19 , Inmunidad Humoral , Vacuna nCoV-2019 mRNA-1273 , Análisis por Matrices de Proteínas , COVID-19/prevención & control , Anticuerpos NeutralizantesRESUMEN
OBJECTIVE: The potential mechanisms linking social participation and depressive symptoms in Chinese individuals with multimorbidity are not yet fully understood. This study aims to explore how cognitive function and activities of daily living (ADLs) mediate the relationship between social participation and depressive symptoms in individuals with multimorbidity. METHODS: We selected 3782 participants with multimorbidity from the 2018 China Health and Retirement Longitudinal Study. Data related to social participation, cognitive function, ADLs, and depressive symptoms were extracted. Regression and Bootstrap analyses were used to explore the sequential mediating effects of social participation, cognitive function, ADLs, and depressive symptoms. RESULTS: (1) There was a significant correlation between social participation, cognitive function, activities of daily living, and depressive symptoms (p < 0.01). (2) Social participation directly affected depressive symptoms (ß = -0.205, p < 0.05). (3) Cognitive function (ß = -0.070, p < 0.01) and activities of daily living (ß = -0.058, p < 0.01) played separate mediating roles in the effect of social participation on depressive symptoms. (4) Cognitive function and activities of daily living had a chain-mediated role in the relationship between social participation and depressive symptoms in patients with multimorbidity (ß = -0.020, p < 0.01). CONCLUSION: A chained mediating effect was found between cognitive function, ADLs, social participation, and depressive symptoms in patients with multimorbidity. Social participation was found to improve the cognitive function of patients with multimorbidity, which in turn enhanced their daily life activities and ultimately alleviated their depressive symptoms.
Asunto(s)
Actividades Cotidianas , Cognición , Depresión , Multimorbilidad , Participación Social , Humanos , Actividades Cotidianas/psicología , Participación Social/psicología , Masculino , Femenino , Depresión/epidemiología , Depresión/psicología , Anciano , China/epidemiología , Estudios Longitudinales , Persona de Mediana Edad , Anciano de 80 o más AñosRESUMEN
BACKGROUND: Chemotherapy resistance is the major cause of recurrence in patients with colorectal cancer (CRC). A previous study found that Fusobacterium (F.) nucleatum promoted CRC chemoresistance. Additionally, metformin rescued F. nucleatum-induced tumorigenicity of CRC. Here, we aimed to investigate whether metformin could revert F. nucleatum-induced chemoresistance and explore the mechanism. METHODS: The role of metformin in F. nucleatum-infected CRC cells was confirmed using cell counting kit 8 assays and CRC xenograft mice. Stemness was identified by tumorsphere formation. Bioinformatic analyses were used to explore the regulatory molecules involved in metformin and F. nucleatum-mediated regulation of the sonic hedgehog pathway. RESULTS: We found that metformin abrogated F. nucleatum-promoted CRC resistance to chemotherapy. Furthermore, metformin attenuated F. nucleatum-stimulated stemness by inhibiting sonic hedgehog signaling. Mechanistically, metformin diminished sonic hedgehog signaling proteins by targeting the MYC/miR-361-5p cascade to reverse F. nucleatum-induced stemness, thereby rescuing F. nucleatum-triggered chemoresistance in CRC. CONCLUSIONS: Metformin acts on F. nucleatum-infected CRC via the MYC/miR-361-5p/sonic hedgehog pathway cascade, subsequently reversing stemness and abolishing F. nucleatum-triggered chemoresistance. Our results identified metformin intervention as a potential clinical treatment for patients with chemoresistant CRC with high amounts of F. nucleatum.
Asunto(s)
Neoplasias Colorrectales , MicroARNs , Humanos , Animales , Ratones , MicroARNs/genética , MicroARNs/metabolismo , Proteínas Hedgehog/genética , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/metabolismo , Fusobacterium nucleatum , Resistencia a Antineoplásicos/genéticaRESUMEN
Dengue is a viral disease transmitted by Aedes aegypti mosquitoes. According to the World Health Organization, about half of the world's population is at risk of dengue. There are four serotypes of the dengue virus. After infection with one serotype, it will be immune to such a serotype. However, subsequent infection with other serotypes will increase the risk of severe outcomes, e.g., dengue hemorrhagic fever, dengue shock syndrome, and even death. Since severe dengue is challenging to predict and lacks molecular markers, we aim to build a multiplexed Flavivirus protein microarray (Flaviarray) that includes all of the common Flaviviruses to profile the humoral immunity and cross-reactivity in the dengue patients with different outcomes. The Flaviarrays we fabricated contained 17 Flavivirus antigens with high reproducibility (R-square = 0.96) and low detection limits (172-214 pg). We collected serums from healthy subjects (n = 36) and dengue patients within 7 days after symptom onset (mild dengue (n = 21), hospitalized nonsevere dengue (n = 29), and severe dengue (n = 36)). After profiling the serum antibodies using Flaviarrays, we found that patients with severe dengue showed higher IgG levels against multiple Flavivirus antigens. With logistic regression, we found groups of markers with high performance in distinguishing dengue patients from healthy controls as well as hospitalized from mild cases (AUC > 0.9). We further reported some single markers that were suitable to separate dengue patients from healthy controls (AUC > 0.9) and hospitalized from mild outcomes (AUC > 0.8). Together, Flaviarray is a valuable tool to profile antibody specificities, uncover novel markers for decision-making, and shed some light on early preventions and treatments.
Asunto(s)
Virus del Dengue , Dengue , Flavivirus , Dengue Grave , Animales , Humanos , Dengue/diagnóstico , Anticuerpos Antivirales , Análisis por Matrices de Proteínas , Reproducibilidad de los Resultados , Antígenos ViralesRESUMEN
OBJECTIVE: The aim of this pilot study was to evaluate the efficacy and safety of intravenous use esomeprazole, metronidazole, and/or levofloxacin in the treatment of Helicobacter pylori (H. pylori)-associated peptic ulcer complications. METHODS: Inpatients with peptic ulcer complications who were not able to take oral medicine were randomly assigned to three groups: triple therapy (esomeprazole, levofloxacin, metronidazole) and dual therapy (esomeprazole, levofloxacin/metronidazole) for 7 days. After intravenous treatment, all patients received open-label oral esomeprazole 20 mg bid for another 1 month. All subjects were followed up for gastroscopy at the seventh day of intravenous treatment to confirm the ulcer healing and 13 C-urea breath test to confirm successful H. pylori eradication 4-6 weeks after completion of oral esomeprazole therapy. RESULTS: The H. pylori eradication rate of both LEV-dual therapy (33.3%, 95% CI: 9.7%-70.0%) and MTZ-dual therapy (50%, 95% CI: 21.5%-78.5%) was significantly lower than that of triple therapy (95%, 95% CI: 71.1%-97.4%) (p = .003, .016). There were no significant differences in the adverse effects among all treatment groups, and the adverse effects were rare. CONCLUSIONS: The intravenous triple regimen, consisting of proton-pump inhibitor, metronidazole, and levofloxacin, could be considered in patients of H. pylori-associated peptic ulcer complications if oral medicine cannot be provided.
Asunto(s)
Infecciones por Helicobacter , Helicobacter pylori , Úlcera Péptica , Humanos , Amoxicilina/uso terapéutico , Antibacterianos/uso terapéutico , Quimioterapia Combinada , Esomeprazol/uso terapéutico , Infecciones por Helicobacter/tratamiento farmacológico , Levofloxacino/farmacología , Metronidazol/uso terapéutico , Úlcera Péptica/tratamiento farmacológico , Proyectos PilotoRESUMEN
Kawasaki disease (KD) is a form of acute systemic vasculitis syndrome that predominantly occurs in children under the age of 5 years. Its etiology has been postulated due to not only genetic factors but also the presence of foreign antigens or infectious agents. To evaluate possible associations between Kawasaki disease (KD) and COVID-19, we investigated humoral responses of KD patients against S-protein variants with SARS-CoV-2 variant protein microarrays. In this study, plasma from a cohort of KD (N = 90) and non-KD control (non-KD) (N = 69) subjects in categories of unvaccinated-uninfected (pre-pandemic), SARS-CoV-2 infected (10-100 days after infection), and 1-dose, 2-dose, and 3-dose BNT162b2 vaccinated (10-100 days after vaccination) was collected. The principal outcomes were non-KD-KD differences for each category in terms of anti-human/anti-His for binding antibodies and neutralizing percentage for surrogate neutralizing antibodies. Binding antibodies against spikes were lower in the KD subjects with 1-dose of BNT162b2, and mean differences were significant for the P.1 S-protein (non-KD-KD, 3401; 95% CI, 289.0 to 6512; P = 0.0252), B.1.617.2 S-protein (non-KD-KD, 4652; 95% CI, 215.8 to 9087; P = 0.0351) and B.1.617.3 S-protein (non-KD-KD, 4874; 95% CI, 31.41 to 9716; P = 0.0477). Neutralizing antibodies against spikes were higher in the KD subjects with 1-dose of BNT162b2, and mean percentage differences were significant for the 1-dose BNT162b2 B.1.617.3 S-protein (non-KD-KD, -22.89%; 95% CI, -45.08 to -0.6965; P = 0.0399), B.1.1.529 S-protein (non-KD-KD, -25.96%; 95% CI, -50.53 to -1.376; P = 0.0333), BA.2.12.1 S-protein (non-KD-KD, -27.83%; 95% CI, -52.55 to -3.115; P = 0.0195), BA.4 S-protein (non-KD-KD, -28.47%; 95% CI, -53.59 to -3.342; P = 0.0184), and BA.5 S-protein (non-KD-KD, -30.42%; 95% CI, -54.98 to -5.869; P = 0.0077). In conclusion, we have found that KD patients have a comparable immunization response to healthy individuals to SARS-CoV-2 infection and COVID-19 immunization.
Asunto(s)
COVID-19 , Síndrome Mucocutáneo Linfonodular , Niño , Humanos , Preescolar , SARS-CoV-2/genética , Síndrome Mucocutáneo Linfonodular/diagnóstico , Síndrome Mucocutáneo Linfonodular/genética , Vacuna BNT162 , Análisis por Matrices de Proteínas , Vacunación , Inmunización , Anticuerpos Neutralizantes , Anticuerpos AntiviralesRESUMEN
The disease progression of COVID-19 varies from mild to severe, even death. However, the link between COVID-19 severities and humoral immune specificities is not clear. Here, we developed a multiplexed spike variant protein microarray (SVPM) and utilized it for quantifying neutralizing activity, drug screening, and profiling humoral immunity. First, we demonstrated the competition between antispike antibody and ACE2 on SVPM for measuring the neutralizing activity against multiple spike variants. Next, we collected the serums from healthy subjects and COVID-19 patients with different severities and profile the neutralizing activity as well as antibody isotypes. We identified the inhibition of ACE2 binding was stronger against multiple variants in severe compared to mild/moderate or critical patients. Moreover, the serum IgG against nonstructural protein 3 was elevated in severe but not in mild/moderate and critical cases. Finally, we evaluated two ACE2 inhibitors, Ramipril and Perindopril, and found the dose-dependent inhibition of ACE2 binding to all the spike variants except for B.1.617.3. Together, the SVPM and the assay procedures provide a tool for profiling neutralizing antibodies, antibody isotypes, and reagent specificities.
Asunto(s)
COVID-19 , Análisis por Matrices de Proteínas , Enzima Convertidora de Angiotensina 2 , Anticuerpos Neutralizantes , Humanos , Isotipos de InmunoglobulinasRESUMEN
BACKGROUND AND AIMS: The recruitment and activation of inflammatory cells in the liver delineates the transition from hepatic steatosis to steatohepatitis (SH). APPROACH AND RESULTS: We found that in SH, γδT cells are recruited to the liver by C-C chemokine receptor (CCR) 2, CCR5, and nucleotide-binding oligomerization domain-containing protein 2 signaling and are skewed toward an interleukin (IL)-17A+ phenotype in an inducible costimulator (ICOS)/ICOS ligand-dependent manner. γδT cells exhibit a distinct Vγ4+ , PD1+ , Ly6C+ CD44+ phenotype in SH. Moreover, γδT cells up-regulate both CD1d, which is necessary for lipid-based antigens presentation, and the free fatty acid receptor, CD36. γδT cells are stimulated to express IL-17A by palmitic acid and CD1d ligation. Deletion, depletion, and targeted interruption of γδT cell recruitment protects against diet-induced SH and accelerates disease resolution. CONCLUSIONS: We demonstrate that hepatic γδT cells exacerbate SH, independent of IL-17 expression, by mitigating conventional CD4+ T-cell expansion and modulating their inflammatory program by CD1d-dependent vascular endothelial growth factor expression.
Asunto(s)
Inmunidad Adaptativa/fisiología , Hígado Graso/etiología , Inmunidad Innata/fisiología , Linfocitos Intraepiteliales/fisiología , Animales , Femenino , Masculino , RatonesRESUMEN
Six parts of lotus (seeds, leaves, plumule, stamens, receptacles and rhizome nodes) are herbal medicines that are listed in the Chinese Pharmacopoeia. Their indications and functions have been confirmed by a long history of clinical practice. To fully understand the material basis of clinical applications, UPLC-QToF-MS combined with the UNIFI platform and multivariate statistical analysis was used in this study. As a result, a total of 171 compounds were detected and characterized from the six parts, and 23 robust biomarkers were discovered. The method can be used as a standard protocol for the direct identification and prediction of the six parts of lotus. Meanwhile, these discoveries are valuable for improving the quality control method of herbal medicines. Most importantly, this was the first time that alkaloids were detected in the stamen, and terpenoids were detected in the cored seed. The stamen is a noteworthy part because it contains the greatest diversity of flavonoids and terpenoids, but research on the stamen is rather limited.
Asunto(s)
Flavonoides/análisis , Lotus/química , Terpenos/análisis , Cromatografía Líquida de Alta Presión , Espectrometría de MasasRESUMEN
BACKGROUND: Splicing factor SRSF3 is an oncogene and overexpressed in various kinds of cancers, however, the function and mechanism involved in colorectal cancer (CRC) remained unclear. The aim of this study was to explore the relationship between SRSF3 and carcinogenesis and progression of CRC. METHODS: The expression of SRSF3 in CRC tissues was detected by immunohistochemistry. The proliferation and invasion rate was analyzed by CCK-8 assay, colony formation assay, transwell invasion assay and xenograft experiment. The expression of selected genes was detected by western blot or real time PCR. RESULTS: SRSF3 is overexpressed in CRC tissues and its high expression was associated with CRC differentiation, lymph node invasion and AJCC stage. Upregulation of SRSF3 was also associated with shorter overall survival. Knockdown of SRSF3 in CRC cells activated ArhGAP30/Ace-p53 and decreased cell proliferation, migration and survival; while ectopic expression of SRSF3 attenuated ArhGAP30/Ace-p53 and increases cell proliferation, migration and survival. Targeting SRSF3 in xenograft tumors suppressed tumor progression in vivo. CONCLUSIONS: Taken together, our data identify SRSF3 as a regulator for ArhGAP30/Ace-p53 in CRC, and highlight potential prognostic and therapeutic significance of SRSF3 in CRC.
RESUMEN
BACKGROUND: Cluster of differentiation 24 (CD24) has recently been reported as a biomarker for colorectal cancer. However, the clinical and prognostic significance of CD24 in colorectal cancer remains controversial. Therefore, we performed a meta-analysis to clarify this issue. METHODS: A comprehensive literature search was performed using Medline, Embase, Web of Science, and CNKI, and the statistical analysis was conducted using Stata software. RESULTS: A total of thirteen studies including 2,180 cases were included in this meta-analysis. The pooled analysis indicated that CD24 expression was associated with lymph node invasion (RR = 0.71 (negative versus positive), 95% CI = 0.52 - 0.96, p = 0.02, Figure 3), differentiation (RR = 0.81 (well versus poor), 95% CI = 0.67 - 0.99, p = 0.04), and T stage (RR = 0.74 (T1 + T2 versus T3 + T4), 95% CI = 0.65 - 0.85, p = 0.00). The prognosis analysis also suggested CD24 overexpression indicating poorer 5-year OS rate (RR = 0.74, 95% CI = 0.58 - 0.93, p = 0.01) However, CD24 was not associated with other clinicopathological features such as tumor size, tumor grade, distant metastasis, TNM stage and Dukes stage. CONCLUSIONS: Taken together, this meta-analysis suggested that CD24 is an efficient prognostic factor in colorectal cancer.
Asunto(s)
Biomarcadores de Tumor/genética , Antígeno CD24/genética , Neoplasias Colorrectales/genética , Ganglios Linfáticos/metabolismo , Biomarcadores de Tumor/metabolismo , Antígeno CD24/metabolismo , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Humanos , Ganglios Linfáticos/patología , Metástasis Linfática , Clasificación del Tumor , Invasividad Neoplásica , Estadificación de Neoplasias , PronósticoRESUMEN
The function and clinical implication of ArfGAP with SH3 domain, ankyrin repeat, and PH domain 3 (ASAP3) in colorectal cancer (CRC) remains undefined. In the present study, we showed that the expression level of ASAP3 was dramatically increased in CRC and its upregulation was associated with American Joint Committee on Cancer stage (P < 0.001) and poor prognosis (P = 0.0022). The combination of stage and ASAP3 expression improved the prediction of survival in CRC patients. Suppression of ASAP3 inhibited cell proliferation by inducing G1 phase arrest without influencing apoptosis. ASAP3 promoted growth of colon tumors in mice with colitis, and accelerated cell invasion and migration in vitro. Increased ASAP3 was associated with activation of the nuclear factor-κB (NF-κB) canonical pathway in CRC. Upregulation of ASAP3 increased the phosphorylation and nuclear translocation of the p65 NF-κB subunit. Mechanistically, ASAP3 interacts with NF-κB essential modulator (NEMO) and could reduce the polyubiquitinylation of NEMO. Overall, ASAP3 might regulate NF-κB via binding to NEMO. ASAP3 acts as an oncogene in colonic cancer and could be a potential biomarker of colon carcinogenesis.
Asunto(s)
Colitis/complicaciones , Neoplasias Colorrectales/patología , Proteínas Activadoras de GTPasa/genética , Quinasa I-kappa B/metabolismo , FN-kappa B/metabolismo , Regulación hacia Arriba , Anciano , Animales , Apoptosis , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Colitis/genética , Neoplasias Colorrectales/genética , Femenino , Proteínas Activadoras de GTPasa/metabolismo , Regulación Neoplásica de la Expresión Génica , Humanos , Masculino , Ratones Noqueados , Persona de Mediana Edad , Trasplante de Neoplasias , Pronóstico , Transducción de Señal , Análisis de Supervivencia , UbiquitinaciónRESUMEN
BACKGROUND: The prevalence of depression among college students is higher than that of the general population. Although a growing body of research suggests that depression in college students and their potential risk factors, few studies have focused on the correlation between depression and risk factors. This study aims to explore the mediating role of perceived social support and resilience in the relationship between trait coping styles and depression among college students. METHODS: A total of 1262 college students completed questionnaires including the Trait Coping Styles Questionnaire (TCSQ), the Patient Health Questionnaire-9 (PHQ-9), the Perceived Social Support Scale (PSSS), and the Resilience Scale-14 (RS-14). Common method bias tests and spearman were conducted, then regressions and bootstrap tests were used to examine the mediating effects. RESULTS: In college students, there was a negative correlation between perceived control PC and depression, with a significant direct predictive effect on depression (ß = -0.067, P < 0.01); in contrast, negative control NC showed the opposite relationship (ß = 0.057, P < 0.01). PC significantly positively predicted perceived social support (ß = 0.575, P < 0.01) and psychological resilience (ß = 1.363, P < 0.01); conversely, NC exerted a significant negative impact. Perceived social support could positively predict psychological resilience (ß = 0.303, P < 0.01), and both factors had a significant negative predictive effect on depression. Additionally, Perceived social support and resilience played a significant mediating role in the relationship between trait coping styles and depression among college students, with three mediating paths: PC/NC â perceived social support â depression among college students (-0.049/0.033), PC/NCâ resilience â depression among college students (-0.122/-0.021), and PC/NC â perceived social support â resilience â depression among college students (-0.016/0.026). CONCLUSION: The results indicate that trait coping styles among college students not only directly predict lower depression but also indirectly influence them through perceived social support and resilience. This suggests that guiding students to confront and solve problems can alleviate their depression.
Asunto(s)
Adaptación Psicológica , Depresión , Resiliencia Psicológica , Apoyo Social , Estudiantes , Humanos , Estudiantes/psicología , Estudiantes/estadística & datos numéricos , Masculino , Femenino , Adulto Joven , Universidades , Depresión/psicología , Adulto , Encuestas y Cuestionarios , Adolescente , Bienestar PsicológicoRESUMEN
The progression of colorectal carcinoma (CRC) to invasive and metastatic disease may involve localized occurrences of epithelial-mesenchymal transition (EMT). However, mechanisms of the EMT process in CRC progression are not fully understood. We previously showed that knockdown of signal transducer and activator of transcription 3 (STAT3) up-regulated E-cadherin (a key component in EMT progression) in CRC. In this study, we examined the roles of STAT3 in CRC EMT and ZEB1, an EMT inducer, in STAT3-induced down-regulation of E-cadherin. Knockdown of STAT3 significantly increased E-cadherin and decreased N-cadherin and vimentin expressions in highly invasive LoVo CRC cells. Meanwhile, overexpression of STAT3 significantly reduced E-cadherin and enhanced N-cadherin and vimentin expressions in weakly invasive SW1116 CRC cells. Activation of STAT3 significantly increased CRC cell invasiveness and resistance to apoptosis. Knockdown of STAT3 dramatically enhanced chemosensitivity of CRC cells to fluorouracil. STAT3 regulated ZEB1 expression in CRC cells, and the STAT3-induced decrease in E-cadherin and cell invasion depended on activation of ZEB1 in CRC cells. Additionally, pSTAT3(Tyr-705) and ZEB1 expressions were significantly correlated with TNM (tumor, lymph node, and metastasis stages) (p < 0.01). In conclusion, STAT3 may directly mediate EMT progression and regulate ZEB1 expression in CRC. ZEB1 may participate in STAT3-induced cell invasion and E-cadherin down-regulation in CRC cells. The expressions of pSTAT3(Tyr-705) and ZEB1 may be positively associated with CRC metastasis. Our data may provide potential targets to prevent and/or treat CRC invasion and metastasis.
Asunto(s)
Cadherinas/genética , Cadherinas/metabolismo , Neoplasias Colorrectales/patología , Regulación hacia Abajo , Transición Epitelial-Mesenquimal , Proteínas de Homeodominio/metabolismo , Factor de Transcripción STAT3/metabolismo , Factores de Transcripción/metabolismo , Apoptosis , Secuencia de Bases , Línea Celular Tumoral , Neoplasias Colorrectales/genética , Proteínas de Homeodominio/genética , Humanos , Quinasas Janus/metabolismo , Datos de Secuencia Molecular , Invasividad Neoplásica , Metástasis de la Neoplasia , Fosfoproteínas/metabolismo , Transducción de Señal , Factores de Transcripción/genética , Vimentina/genética , Vimentina/metabolismo , Homeobox 1 de Unión a la E-Box con Dedos de ZincRESUMEN
The continuous mutation of SARS-CoV-2 highlights the need for rapid, cost-effective, and high-throughput detection methods. To better analyze the antibody levels against SARS-CoV-2 and its variants in vaccinated or infected subjects, we developed a multiplex detection named Barcode Bead Fluorescence (BBF) assay. These barcode beads were magnetic, characterized by 2-dimensional edges, highly multiplexed, and could be decrypted with visible light. We conjugated 12 magnetic barcode beads with corresponding nine spike proteins (wild-type, alpha, beta, gamma, delta, and current omicrons), two nucleocapsid proteins (wild-type and omicron), and one negative control. First, the conjugated beads underwent serial quality controls via fluorescence labeling, e.g., reproducibility (R square = 0.99) and detection limits (119 pg via anti-spike antibody). Next, we investigated serums from vaccinated subjects and COVID-19 patients for clinical applications. A significant reduction of antibody levels against all variant beads was observed in both vaccinated and COVID-19 studies. Subjects with two doses of mRNA-1273 exhibited the highest level of antibodies against all spike variants compared to two doses of AZD1222 and unvaccinated. We also found that COVID-19 patients showed higher antibody levels against spike beads from wild-type, alpha, beta, and delta. Finally, the nucleocapsid beads served as markers to distinguish infections from vaccinated subjects. Overall, this study developed the BBF assay for analyzing humoral immune responses, which has the advantages of robustness, automation, scalability, and cost-effectiveness.
Asunto(s)
Técnicas Biosensibles , COVID-19 , Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , ChAdOx1 nCoV-19 , Reproducibilidad de los Resultados , Anticuerpos AntiviralesRESUMEN
Objectives: RA is an autoimmune disease characterized by chronic inflammation and joint destruction. Biologics are crucial to achieving treat-to-target goals in patients with RA. The global spread and continuous variation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) necessitate the monitoring of variant-specific humoral responses post-vaccination. The aim of this study was to investigate how different biologic treatments for vaccinated RA patients might affect their neutralizing antibodies against multiple SARS-CoV-2 variants. Methods: We recruited RA patients who had received three doses of conventional SARS-CoV-2 vaccines and were treated with various biologics, e.g. TNF inhibitor (etanercept), IL-6 inhibitor (tocilizumab), CTLA4-Ig (abatacept) or anti-CD20 (rituximab). Serum samples were used to profile the binding and neutralizing antibodies using our own SARS-CoV-2 variant (CoVariant) protein array, developed previously. Results: Compared with healthy controls, only RA therapy with rituximab showed a reduction in neutralizing antibodies capable of targeting spike proteins in SARS-CoV-2 wild-type and most variants. This reduction was not observed in binding antibodies against SARS-CoV-2 wild-type or its variants. Conclusion: After receiving three doses of SARS-CoV-2 vaccination, RA patients who underwent rituximab treatment generated sufficient antibodies but exhibited lower neutralizing activities against wild-type and multiple variants, including current Omicron. Other biological DMARDs, e.g. TNF inhibitor, IL-6 inhibitor and CTLA4-Ig, did not show obvious inhibition.
RESUMEN
Colorectal carcinogenesis coincides with immune cell dysfunction. Metformin has been reported to play a role in stimulating antitumor immunity, suggesting it could be used to overcome immunosuppression in colorectal cancer. Herein, using single-cell RNA sequencing (scRNA-seq), we showed that metformin remodels the immune landscape of colorectal cancer. In particular, metformin treatment expanded the proportion of CD8+ T cells and potentiated their function. Analysis of the metabolic activities of cells in the colorectal cancer tumor microenvironment (TME) at a single-cell resolution demonstrated that metformin reprogrammed tryptophan metabolism, which was reduced in colorectal cancer cells and increased in CD8+ T cells. Untreated colorectal cancer cells outcompeted CD8+ T cells for tryptophan, leading to impaired CD8+ T-cell function. Metformin in turn reduced tryptophan uptake by colorectal cancer cells, thereby restoring tryptophan availability for CD8+ T cells and increasing their cytotoxicity. Metformin inhibited tryptophan uptake in colorectal cancer cells by downregulating MYC, which led to a reduction in the tryptophan transporter SLC7A5. This work highlights metformin as an essential regulator of T-cell antitumor immunity by reprogramming tryptophan metabolism, suggesting it could be a potential immunotherapeutic strategy for treating colorectal cancer. SIGNIFICANCE: Analysis of the impact of metformin on the colorectal cancer immunometabolic landscape at a single-cell resolution shows that metformin alters cancer cell tryptophan metabolism to stimulate CD8+ T-cell antitumor activity.
Asunto(s)
Neoplasias Colorrectales , Metformina , Humanos , Metformina/farmacología , Metformina/uso terapéutico , Triptófano , Linfocitos T CD8-positivos , Terapia de Inmunosupresión , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/metabolismo , Microambiente TumoralRESUMEN
Epidemiological studies have indicated an association between statin use and reduced incidence of colorectal cancer (CRC), and work in preclinical models has demonstrated a potential chemopreventive effect. Statins are also associated with reduced dysbiosis in the gut microbiome, yet the role of the gut microbiome in the protective effect of statins in CRC is unclear. Here we validated the chemopreventive role of statins by retrospectively analysing a cohort of patients who underwent colonoscopies. This was confirmed in preclinical models and patient cohorts, and we found that reduced tumour burden was partly due to statin modulation of the gut microbiota. Specifically, the gut commensal Lactobacillus reuteri was increased as a result of increased microbial tryptophan availability in the gut after atorvastatin treatment. Our in vivo studies further revealed that L. reuteri administration suppressed colorectal tumorigenesis via the tryptophan catabolite, indole-3-lactic acid (ILA). ILA exerted anti-tumorigenic effects by downregulating the IL-17 signalling pathway. This microbial metabolite inhibited T helper 17 cell differentiation by targeting the nuclear receptor, RAR-related orphan receptor γt (RORγt). Together, our study provides insights into an anti-cancer mechanism driven by statin use and suggests that interventions with L. reuteri or ILA could complement chemoprevention strategies for CRC.
Asunto(s)
Neoplasias Colorrectales , Inhibidores de Hidroximetilglutaril-CoA Reductasas , Limosilactobacillus reuteri , Microbiota , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/farmacología , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Triptófano , Estudios Retrospectivos , Neoplasias Colorrectales/prevención & controlRESUMEN
Abnormalities in the JAK2/STAT3 pathway are involved in the pathogenesis of colorectal cancer (CRC), including apoptosis. However, the exact mechanism by which dysregulated JAK2/STAT3 signalling contributes to the apoptosis has not been clarified. To investigate the role of both JAK2 and STAT3 in the mechanism underlying CRC apoptosis, we inhibited JAK2 with AG490 and depleted STAT3 with a small interfering RNA. Our data showed that inhibition of JAK2/STAT3 signalling induced CRC cellular apoptosis via modulating the Bcl-2 gene family, promoting the loss of mitochondrial transmembrane potential (Δψm) and the increase of reactive oxygen species. In addition, our results demonstrated that the translocation of cytochrome c (Cyt c), caspase activation and cleavage of poly (ADP-ribose) polymerase (PARP) were present in apoptotic CRC cells after down-regulation of JAK2/STAT3 signalling. Moreover, inhibition of JAK2/STAT3 signalling suppressed CRC xenograft tumour growth. We found that JAK2/STAT3 target genes were decreased; meanwhile caspase cascade was activated in xenograft tumours. Our findings illustrated the biological significance of JAK2/STAT3 signalling in CRC apoptosis, and provided novel evidence that inhibition of JAK2/STAT3 induced apoptosis via the mitochondrial apoptotic pathway. Therefore, JAK2/STAT3 signalling may be a potential target for therapy of CRC.