细胞与分子免疫学杂志最新文献

Objective To express and purify Alicyclobacillus acidiphilus Cas12b (AaCas12b) protein, and establish a rapid clustered regularly interspaced short palindromic repeats (CRISPR)-based assay for Mycobacterium tuberculosis (Mtb) detection, thereby providing a novel tool for clinical diagnosis of tuberculosis (TB). Methods Following construction and transformation of the recombinant expression vector into E. coli BL21 (DE3), soluble expression of the recombinant AaCas12b protein was induced with low-concentration isopropyl β-D-thiogalactopyranoside (IPTG) at low temperature, followed by purification using nickel-affinity chromatography. Subsequently, a specific recombinase polymerase amplification (RPA) assay targeting the IS6110 insertion sequence of Mtb was developed, and the corresponding single guide RNA (sgRNA) was prepared via in vitro transcription. Then a novel CRISPR-AaCas12b detection assay for Mtb was established, and its diagnostic performance was systematically evaluated against conventional clinical methods including acid-fast staining, T-SPOT.TB assay, Mycobacterium culture, and GeneXpert assay. Results A prokaryotic expression and purification system for recombinant AaCas12b was successfully established, yielding AaCas12b protein with activity at a concentration of 13.42 mg/mL and a recovery rate of 61.45%. A specific RPA-CRISPR-AaCas12b detection system was established targeting the IS6110 sequence, with a limit of detection (LOD) of 1.5 CFU/mL for Mtb. In clinical sample validation, the system achieved a sensitivity of 95.83% (95% CI: 79.97%~99.27%), a specificity of 92.31% (95% CI: 75.88%~97.88%), and an accuracy of 94.00% (95% CI: 83.78%~97.93%). Conclusion The established AaCas12b protein preparation system is efficient and stable, enabling the production of low-cost, high-activity tool enzymes. The RPA-CRISPR-AaCas12b TB diagnostic assay targeting the IS6110 sequence exhibits excellent specificity and sensitivity, providing a technical reference and experimental basis for the further development of rapid TB diagnostic platforms.

Inflammatory bowel disease (IBD) belongs to a group of idiopathic intestinal inflammatory disorders that affect the ileum, rectum, and colon. There is still no effective treatment methods, and with the increasing number of patients, IBD has brought a heavy burden to the medical system. Chronic inflammation mediated by abnormal activation of the intestinal mucosal immune system plays an important role in the occurrence and development of IBD. As a common intracellular sensor that activates the innate immune system, inflammasomes have a signaling cascade regulated by ubiquitination/deubiquitination. Multiple studies have confirmed that NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is continuously activated in the progression of IBD, therefore targeting this oligomeric protein complex is of great significance for the treatment of IBD. This review systematically introduces the activation mechanism of NLRP3 inflammasome, the regulation of NLRP3 signaling cascade by ubiquitination/deubiquitination modification, and the role of NLRP3 inflammasome in the progression of IBD under the post-translational modification regulation. It also systematically discusses the feasibility of targeting this protein complex to provide new ideas for the treatment of IBD.

Objective To investigate the regulatory role of H2-Eb1 gene in allergic rhinitis (AR) under plateau environments and elucidate the differential effects of acute plateau exposure and long-term plateau adaptation on AR, so as to provide evidence for the prevention and treatment of AR in plateau regions. Methods The establishment of an AR model was conducted through ovalbumin (OVA) intraperitoneal injection and nasal drip methods. Male C57BL/6 mice aged 6-8 weeks were divided into the following groups: plain normal group, plain AR group, plateau normal group, plateau AR group, acute plateau exposure normal group, and acute plateau exposure AR group, with 10 mice in each group. The plain groups were maintained at an altitude of 1500 m, the plateau groups were housed in a hyperbaric chamber where an altitude of 7000 m was simulating, and the acute plateau exposure groups underwent sensitization treatment first and were then transferred to the hyperbaric chamber for 1 week. Rhinitis symptoms, nasal mucosa pathology, H2-Eb1 mRNA and protein expression in mice were observed and evaluated. Results Rhinitis symptoms were exhibited by all AR groups, with the most severe symptoms being observed in the acute plateau exposure AR group and the mildest symptoms being found in the long-term plateau AR group. Furthermore, decreased H2-Eb1 protein expression and improved inflammatory infiltration were observed in the plateau AR group, whereas the opposite pattern was exhibited by the acute plateau exposure AR group. Conclusion Acute plateau exposure is shown to upregulate H2-Eb1 protein expression, with the acute plateau response being thereby aggravated; in contrast, downregulation of the protein expression of this gene is achieved through long-term adaptation to plateau environments, suggesting that H2-Eb1 serves as a crucial regulatory target for modulating acute plateau response.

Following Mycobacterium tuberculosis (Mtb) infection, the pathogen can establish a metabolically quiescent or replication-arrested dormant state within the host, leading to asymptomatic latent tuberculosis infection (LTBI). Under specific conditions, these dormant bacilli may reactivate and replicate, resulting in symptomatic clinical infection and constituting a major reservoir for active tuberculosis (TB). Dormant Mtb exhibits reduced susceptibility to conventional anti-TB drugs, and current TB vaccines fail to prevent the establishment or reactivation of dormant Mtb, rendering latent infection a major challenge in TB control. The formation of Mtb dormancy requires a unique microenvironment characterized by hypoxia, nutrient deprivation, and host immunosuppression. As an intracellular pathogen, Mtb primarily infects macrophages. Recent studies have revealed that Mtb can also infect mesenchymal stem cells (MSCs), where it persists in a dormant state for a long term. MSCs exhibit a microenvironment marked by hypoxia, cellular quiescence, lipid droplet biogenesis, and elevated autophagic activity. Moreover, MSCs exert immunosuppressive effects on the host immune response to Mtb, thereby facilitating the formation of bacterial dormancy and long-term intracellular survival, which further contributes to TB reactivation and relapse. This review summarizes current advances in understanding Mtb infection of MSCs and the underlying mechanisms associated with Mtb dormancy within this cellular niche.

Objective To investigate the heterogeneity characteristics of cancer-associated fibroblasts (CAF) in acral melanoma (AM) and their functional roles in the tumor microenvironment (TME), providing a theoretical basis for developing effective therapeutic strategies. Methods The single-cell RNA sequencing data used in this study were previously generated by our research group and have been deposited in the HRA001804 dataset of the National Genomics Data Center (NGDC). Five AM patient samples were selected from this dataset for analysis, including 4 primary tumor samples (PL1, PL2, PL4, PL5) and 1 lymph node metastasis sample (LG2). Bioinformatics methods including unsupervised clustering, pseudotime trajectory analysis, transcription factor regulatory network analysis, and cell-cell communication analysis were used for data mining. Results Three subtypes of CAF were identified from 41 960 high-quality cells: immunomodulatory CAF (iCAF), myofibroblastic CAF (mCAF), and proliferative CAF (pCAF). Pseudotime trajectory analysis demonstrated that mCAF were located at the initiation of differentiation, while iCAF and pCAF were distributed in distinct terminal branches. FOS-like 1, AP-1 transcription factor subunit (FOSL1) was identified as a key transcription factor regulating the differentiation of CAF into pCAF, and its co-expression modules were enriched in the P53 signal pathway, AP-1 transcription factor network, and MYC-mediated cell proliferation pathways. Cell communication analysis revealed an interaction network centered on iCAF, with critical pathways including CSPG4-(ITGA2+ITGB1), GDF15-TGFBR2, LGALS9-CD45/CD44, and CD70-CD27 signal pathways. Conclusion CAF in AM exhibits high heterogeneity, with FOSL1 playing a key regulatory role in CAF differentiation. CAF participates in tumor microenvironment regulation through complex intercellular communication networks. These findings provide important insights into understanding the biological characteristics of acral melanoma and developing targeted therapeutic strategies.

The neutrophil-to-lymphocyte ratio (NLR), as a simple and reliable biomarker of systemic inflammation, has garnered significant attention in the clinical management of various inflammatory diseases and malignancies. By quantifying the relative counts of neutrophils and lymphocytes, this parameter reflects the body's immune-inflammatory status. This article provides a comprehensive review of the current research on the role of NLR in chronic obstructive pulmonary disease (COPD) concomitant with advanced lung cancer (LC) and aims to offer insights into novel diagnostic approaches, prognostic evaluation strategies, and potential therapeutic implications for clinical practice.

Objective To investigate the effect of histone deacetylase 11 (HDAC11) on oxidized low density lipoprotein (ox-LDL)-induced mouse macrophage foam cell formation. Methods Macrophages derived from the RAW264.7 cell line were divided into three groups: Control group, ox-LDL group, and ox-LDL combined with Rapamycin group. Oil red O staining was used to observe the formation of foam cells, the content of total cholesterol (TC) and triglyceride (TG) in macrophages were measured using microplate reader. The expression of HDAC11, microtubule-associated protein 1 light chain 3-β (LC3B) II and p62 were detected by Western blot assay. Si-HDAC11 was transfected into macrophages, and qRT-PCR and Western blot assay were employed to measure the interference efficiency of HDAC11. On the basis of ox-LDL treatment, the expression of HDAC11 in RAW264.7 cells was knock down. Oil red O staining was used to observe the lipid droplet content, the content of TC and TG were detected by using microplate reader, and LC3BII and p62 protein expressions were detected by Western blot assay. Results Compared to the control group, the protein expression of LC3BII was reduced, while p62 and HDAC11 protein were increased, which is accompanied by a significant increase of the lipid droplet content, TC and TG. Simultaneously, treated RAW264.7 cells with Rapamycin, an autophagy agonist, LC3BII and p62 protein expressions, as well as lipid droplets, TC, and TG content changes were significantly alleviated in RAW264.7 cells. LC3BII expression was significantly increased after the transfection of si-HDAC11 into macrophages, while p62 protein expression was decreased, and intracellular lipid droplets, TC and TG contents were also reduced correspondingly. Conclusion HDAC11 promotes ox-LDL-induced mouse macrophage foam cell formation via inhibiting autophagy.

Red blood cell alloimmunity is an immune response triggered by individuals who contact with red blood cells different from their own blood type antigens during blood transfusion or pregnancy, which can cause cross-matching incompatibility, hemolytic transfusion reaction and hemolytic disease of newborns. Human leukocyte antigen (HLA) gene polymorphism plays an important role in the alloimmune response induced by the blood type antigen on erythrocytes. This review introduces the research status and progress of the genetic background of HLA gene polymorphism and multiple erythrocytic blood type system antigens such as Rh, Duffy, Kell, Kidd and MNS in different regions and races. Among them, the alloimmunity of Rh blood type system antigen has attracted much attention.

Sphingosine-1-Phosphate (S1P) is a bioactive phospholipid molecule that regulates multiple signaling pathways through its interaction with sphingosine-1-phosphate receptors (S1PRs). In cancer tissues, S1P is not only associated with the survival of cancer cells, but also related to the angiogenesis, inflammatory responses, and immune evasion in the tumor microenvironment (TME). Various inhibitors of the S1P kinase have shown significant anti-tumor effects in pre-clinical studies by directly inhibiting tumor growth and metastasis, and enhancing the therapeutic potential of both chemotherapy and immunotherapy. Nevertheless, the complex and pleiotropic nature of the S1P signaling pathway presents considerable challenges for the development of targeted therapeutic strategies. This paper reviews the biological functions and research progress of S1P and its related pathways in cancer, offering novel insights and potential directions for cancer therapy.

Sepsis is one of the major diseases threatening human health. Under inflammatory stress, the rapid activation of glucocorticoids (GCs) and catecholamines (CAs) in the adrenal gland is crucial for maintaining the body's homeostasis. During severe inflammation, impairment of the hypothalamic-pituitary-adrenal (HPA) axis leads to suppressed secretion of corticotropin-releasing hormone (CRH) and adrenocorticotropic hormone (ACHT). However, studies reveal that the systemic glucocorticoid levels can remain normal or even elevated, suggesting a transition in the stress response mechanism from the central, pituitary-dependent regulatory system to local, pituitary-independent regulatory mechanisms. Among the various potential mechanisms, activation of the local adrenal microenvironment plays a significant role. In this microenvironment, interactions between adrenal cortical cells, medullary cells, vascular endothelial cells, and various immune cells contribute to the sustained production of GCs, which is pivotal in maintaining the internal homeostasis. This review focuses on the role of the local adrenal microenvironment in regulating the secretion of GCs and CAs, key hormones for maintaining homeostasis under inflammatory stress, with the aim of providing new insights into the etiology and pathogenesis of adrenal diseases.