Journal of Leukocyte Biology最新文献_第7页

Development of 2LTRZFP-expressing induced pluripotent stem cells as a potential anti-HIV-1 gene therapy against viral integration. 开发表达 2LTRZFP 的诱导多能干细胞，作为抗 HIV-1 病毒整合的潜在基因疗法。

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiaf018

Kritayaporn Saiprayong, Koollawat Chupradit, Pasut Sasithong, Siriwal Suwanpitak, Saitong Muneekaew, Nontaphat Thongsin, Jakkrapatra Srisantitham, Methichit Wattanapanitch

{"title":"Development of 2LTRZFP-expressing induced pluripotent stem cells as a potential anti-HIV-1 gene therapy against viral integration.","authors":"Kritayaporn Saiprayong, Koollawat Chupradit, Pasut Sasithong, Siriwal Suwanpitak, Saitong Muneekaew, Nontaphat Thongsin, Jakkrapatra Srisantitham, Methichit Wattanapanitch","doi":"10.1093/jleuko/qiaf018","DOIUrl":"10.1093/jleuko/qiaf018","url":null,"abstract":"Highly active antiretroviral drug is the standard treatment for HIV-1 infection to suppress the viral load. However, this treatment does not completely eradicate the virus; it simply decreases the viral load to undetectable levels. The development of a novel therapy to cure the disease is essential. Previously, we developed an engineered zinc finger protein (ZFP) that specifically binds to the 2-LTR-circle junction (2LTRZFP), the target site for viral integrase, preventing HIV-1 integration in human CD34+ hematopoietic stem/progenitor cells (HSPCs) and macrophages. Although the transduction efficiency of 2LTRZFP was ∼50%, purifying and expanding the 2LTRZFP-expressing HSPCs proved difficult. In addition, the batch-to-batch variability in transduction efficiency could have a major impact on the therapeutic efficacy. In this study, we introduced the 2LTRZFP into human induced pluripotent stem cells (iPSCs) followed by clonal isolation and functional validation of the 2LTRZFP. Upon the HIV-1 challenge, the 2LTRZFP protein was found to inhibit the viral integration in iPSCs, iPSC-derived HSPCs, and macrophages. The engineered iPSC clone could be differentiated into functional macrophages, as evidenced by M1 and M2 polarization, and phagocytosis. Our finding revealed that the 2LTRZFP did not perturb the macrophage differentiation process. Therefore, the 2LTRZFP-expressing iPSCs could provide an unlimited supply of HIV-1-resistant HSPCs for transplantation, potentially leading to HIV-1-resistant blood cells. The knowledge obtained from this study will provide a cornerstone for HIV-1 gene therapy using HSPC transplantation as a sustainable HIV-1 treatment in the future.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143414335","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Different behavior of NK cells isolated from healthy women and women with recurrent spontaneous abortion after treatment with human amniotic epithelial cells. 人羊膜上皮细胞治疗后健康妇女和复发性自然流产妇女NK细胞的不同行为

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiaf020

Fatemeh Rezayat, Nafiseh Esmaeil, Parvaneh Nikpour, Awat Feizi, Abbas Rezaei

{"title":"Different behavior of NK cells isolated from healthy women and women with recurrent spontaneous abortion after treatment with human amniotic epithelial cells.","authors":"Fatemeh Rezayat, Nafiseh Esmaeil, Parvaneh Nikpour, Awat Feizi, Abbas Rezaei","doi":"10.1093/jleuko/qiaf020","DOIUrl":"https://doi.org/10.1093/jleuko/qiaf020","url":null,"abstract":"Maternal immunotolerance during pregnancy is heavily dependent on the critical properties of human amniotic epithelial cells (hAECs). Recurrent spontaneous abortion (RSA) is one of the most common diseases in women and is caused by feto-maternal immunotolerance disruption. The objective of this study is to investigate how hAEECs affect pNK cells isolated from RSA and healthy women in terms of immunomodulation. Peripheral blood NK cells were isolated from 20 women with RSA and 20 healthy women. Purified NK cells were co-cultured with hAECs, obtained from full-term healthy pregnant women at different cellular ratios. After 24 and 72 h of incubation, the expression of immunomodulatory genes in hAECs, immunophenotype, and cytotoxicity of NK cells, and cytokine production were investigated using real-time PCR, flow cytometry, and ELISA techniques, respectively. We observed a significant increase in TGF-β and IL-10 production, and CD56bright CD16+ subpopulation in pNK cells, a significant decrease in IFN-γ production and CD107a and FasL expression on NK cells. Also, NK cells' cytotoxicity against K562 cells was diminished after co-culture with hAECs. The expression of TGF-β and HLA-G genes by hAECs was diminished after co-culture with NK cells isolated from women with RSA. Our research indicates that the interaction between NK cells and hAECs influences the phenotype and function of both cells. Also, NK cells belonging to women with RSA and healthy women exhibit different behavior during treatment with hAECs, possibly due to NK cell dysfunction. However, extensive research is required to assess NK cell defects and their mutual interaction with hAECs.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":"117 4","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144004255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Decoding eosinophils: biological insights hidden in plain sight? 解读嗜酸性粒细胞：隐藏在视线中的生物学见解？

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiaf030

Julia L M Dunn, Lisa A Spencer

引用次数: 0

Enhanced Siglec-8 and HLA-DR and reduced CRTH2 surface expression highlight a distinct phenotypic signature of circulating eosinophils in atopic dermatitis. Siglec-8和HLA-DR的增强以及CRTH2表面表达的减少，突显了特应性皮炎中循环嗜酸性粒细胞的独特表型特征。

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiaf023

Frédéric Dezoteux, Pierre Marcant, Arnaud Dendooven, Émeline Delaunay, Stéphane Esnault, Jacques Trauet, Guillaume Lefèvre, Delphine Staumont-Sallé

{"title":"Enhanced Siglec-8 and HLA-DR and reduced CRTH2 surface expression highlight a distinct phenotypic signature of circulating eosinophils in atopic dermatitis.","authors":"Frédéric Dezoteux, Pierre Marcant, Arnaud Dendooven, Émeline Delaunay, Stéphane Esnault, Jacques Trauet, Guillaume Lefèvre, Delphine Staumont-Sallé","doi":"10.1093/jleuko/qiaf023","DOIUrl":"10.1093/jleuko/qiaf023","url":null,"abstract":"Atopic dermatitis and other type 2 immune response diseases are often linked to elevated eosinophil levels in the blood. Although the role of eosinophils in atopic dermatitis pathophysiology is suspected, it remains unclear. The development of new treatments targeting the type 2 response, particularly cytokines involved in eosinophil activation and chemotaxis, makes it necessary to identify potential eosinophil profiles in atopic dermatitis that may respond to these treatments. A prospective study was conducted comparing blood eosinophil phenotypes in patients with moderate to severe atopic dermatitis (n = 19) without recent systemic treatment to healthy individuals (n = 19). The primary outcome was the membranous phenotypic signature of eosinophils, assessed by flow cytometry. Most patients with atopic dermatitis (84%) had early onset in childhood, a severe disease (mean SCORing Atopic Dermatitis of 57.5), and elevated blood eosinophil counts (310 per µL in atopic dermatitis vs 120 in healthy individuals, P < 0.0001). Patients with atopic dermatitis exhibited lower CRTH2 on eosinophils but higher levels of human leukocyte antigen-DR isotype and Siglec-8 compared to healthy individuals. Other surface proteins showed no significant differences. Clustering analysis confirmed increased Siglec-8 in patients with atopic dermatitis. Additionally, patients with atopic dermatitis had higher serum levels of type 2 immune response markers such as eotaxin-2, IL-5, IL-3, and TARC. Circulating eosinophils in patients with atopic dermatitis show a distinct phenotypic profile, suggesting a role in atopic dermatitis pathophysiology and potential involvement in differential treatment responses.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143492296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-dimensional spectral flow cytometry of activation and phagocytosis by peripheral human polymorphonuclear leukocytes. 人类外周多形核白细胞激活和吞噬的高维光谱流式细胞术。

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiaf025

Evan R Lamb, Ian J Glomski, Taylor A Harper, Michael D Solga, Alison K Criss

{"title":"High-dimensional spectral flow cytometry of activation and phagocytosis by peripheral human polymorphonuclear leukocytes.","authors":"Evan R Lamb, Ian J Glomski, Taylor A Harper, Michael D Solga, Alison K Criss","doi":"10.1093/jleuko/qiaf025","DOIUrl":"10.1093/jleuko/qiaf025","url":null,"abstract":"Polymorphonuclear leukocytes (PMNs) are terminally differentiated phagocytes with pivotal roles in infection, inflammation, tissue injury, and resolution. PMNs display a breadth of responses to diverse endogenous and exogenous stimuli, making understanding of these innate immune responders vital yet challenging to achieve. Here, we report a 22-color spectral flow cytometry panel to profile primary human PMNs for surface marker expression of activation, degranulation, phagocytosis, migration, chemotaxis, and interaction with fluorescently labeled cargo. We demonstrate the surface marker response of PMNs to phorbol ester stimulation compared with untreated controls in an adherent PMN model with additional analysis of intra- and inter-subject variability. PMNs challenged with the Gram-negative bacterial pathogen Neisseria gonorrhoeae revealed infectious dose-dependent changes in surface marker expression in bulk, population-level analysis. Imaging flow cytometry complemented spectral cytometry, demonstrating that fluorescence signal from labeled bacteria corresponded with bacterial burden on a per-cell basis. Spectral flow cytometry subsequently identified surface markers, which varied with direct PMN-bacterium association as well as those which varied in the presence of bacteria but without phagocytosis. This spectral panel protocol highlights best practices for efficient customization and is compatible with downstream approaches such as spectral cell sorting and single-cell RNA-sequencing for applicability to diverse research questions in the field of PMN biology.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12031652/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143557146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Characterizing a low-density neutrophil gene signature in acute and chronic infections and its impact on disease severity. 表征急性和慢性感染中的低密度中性粒细胞基因特征及其对疾病严重程度的影响。

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiaf027

Matheus Aparecido de Toledo, João Victor Souza de Lima, Reinaldo Salomão, Giuseppe G F Leite

{"title":"Characterizing a low-density neutrophil gene signature in acute and chronic infections and its impact on disease severity.","authors":"Matheus Aparecido de Toledo, João Victor Souza de Lima, Reinaldo Salomão, Giuseppe G F Leite","doi":"10.1093/jleuko/qiaf027","DOIUrl":"10.1093/jleuko/qiaf027","url":null,"abstract":"Low-density neutrophils (LDNs) or polymorphonuclear myeloid-derived suppressor cells are involved in the pathogenesis of cancer, autoimmune, and infectious diseases. They are crucial in the host response to invading pathogens, especially during acute illness, and are associated with poor prognosis in many infectious diseases. However, their gene expression profile and contribution to disease outcomes are not well described. We conducted a meta-analysis of gene expression datasets from peripheral blood mononuclear cells (PBMCs), focusing on patients with viral and bacterial infections. We identified a consensus set of 2,798 differentially expressed genes. Among these, 49 genes were commonly found in both the neutrophil degranulation pathway and the granule lumen-specific community. To validate this signature, we evaluated its expression in RNA-seq datasets, finding consistent upregulation of 24 genes in severe infections, 17 of them overlapped with genes overexpressed in CD16int cells. We also investigated the abundance of LDN-related proteins in a PBMC proteomics dataset from a cohort of sepsis and septic shock patients. Out of the 17 genes analyzed, 13 corresponding proteins were identified, 10 of which demonstrated significantly higher abundance in sepsis and septic shock patients compared with healthy controls. In conclusion, our study identified a pattern of 17 upregulated LDN genes, common to PBMC transcriptome and RNA-seq, and upregulated in CD16int, associated with acute infections and severe clinical outcomes, marking the first time these genes have been collectively presented as a potential signature of LDNs in relation to disease severity. Further research with prospective cohorts is needed to validate this LDN signature and explore its clinical implications.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143557129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Depression-like behavior is associated with changes in the meningeal lymphatic vasculature and meningeal B cells in a murine lupus model. 在小鼠狼疮模型中，抑郁样行为与脑膜淋巴血管和脑膜B细胞的变化有关。

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiaf016

Alexandra Olate-Briones, Sofía Albornoz-Muñoz, Francisca Rodriguez-Arriaza, Victor Rojas-Henriquez, Stefanny S Rojas, Noelia Escobedo, Andrés A Herrada

{"title":"Depression-like behavior is associated with changes in the meningeal lymphatic vasculature and meningeal B cells in a murine lupus model.","authors":"Alexandra Olate-Briones, Sofía Albornoz-Muñoz, Francisca Rodriguez-Arriaza, Victor Rojas-Henriquez, Stefanny S Rojas, Noelia Escobedo, Andrés A Herrada","doi":"10.1093/jleuko/qiaf016","DOIUrl":"https://doi.org/10.1093/jleuko/qiaf016","url":null,"abstract":"Meningeal lymphatic vasculature (mLV) comprises a network of vessels responsible for draining immune cells and fluid from the central nervous system (CNS) into the deep cervical lymph nodes. While changes in mLV function have been implicated in several neurodegenerative disorders, its role in autoimmune diseases is less clear. Systemic lupus erythematosus (SLE) is an autoimmune disease affecting multiple organs. When SLE affects the CNS, it is known as neuropsychiatric lupus (NPSLE), although the status of mLV during NPSLE has not been yet evaluated. Here, by using the lupus FcγRIIb-/- murine model, we found that this model develops NPSLE along with increased mLV coverage and function at 4 mo of age. Altered B cell developmental stages were evident in this lupus mouse model. In fact, increased B cell clusters in the meninges of FcγRIIb-/- mice were also observed. These findings suggest that mLV morphology and function are increased in FcγRIIb-/- mice together with changes in the meningeal B cell population that could have an impact on NPSLE symptoms.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":"117 4","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143986187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Protective function of ex vivo-expanded CD8 T cells in a mouse model of adoptive therapy for cytomegalovirus infection depends on integrin beta 1 but not CXCR3, CTLA4, or PD-1 expression. 在巨细胞病毒感染过继治疗的小鼠模型中，体外扩增的CD8 T细胞的保护功能取决于整合素β 1，而不是CXCR3、CTLA4或PD-1的表达。

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiae256

Xiaokun Liu, Rodrigo Gutierrez Jauregui, Yvonne Lueder, Stephan Halle, Laura Ospina-Quintero, Christiane Ritter, Anja Schimrock, Stefanie Willenzon, Anika Janssen, Karen Wagner, Martin Messerle, Berislav Bošnjak, Reinhold Förster

{"title":"Protective function of ex vivo-expanded CD8 T cells in a mouse model of adoptive therapy for cytomegalovirus infection depends on integrin beta 1 but not CXCR3, CTLA4, or PD-1 expression.","authors":"Xiaokun Liu, Rodrigo Gutierrez Jauregui, Yvonne Lueder, Stephan Halle, Laura Ospina-Quintero, Christiane Ritter, Anja Schimrock, Stefanie Willenzon, Anika Janssen, Karen Wagner, Martin Messerle, Berislav Bošnjak, Reinhold Förster","doi":"10.1093/jleuko/qiae256","DOIUrl":"https://doi.org/10.1093/jleuko/qiae256","url":null,"abstract":"The adoptive transfer of virus-specific T cells (VSTs) represents a therapeutic option for viral infection treatment in immunocompromised patients. Before administration, ex vivo culture enables VST expansion. However, it is unclear how ex vivo expansion affects the circulation, homing, and intra-tissue migration of administered VSTs. We established a model of VST immunotherapy of acute cytomegalovirus infection using adoptive transfer of ex vivo-expanded OT-I CD8 T cells (recognizing SIINFEKL peptide) into Rag2-/- mice infected with murine cytomegalovirus (MCMV) encoding for the SIINFEKL peptide. Ex vivo expansion induced an effector T cell phenotype and affected the expression of integrins and chemokine receptors. CRISPR/Cas9-mediated gene deletions enabled us to address the role of selected genes in the homing of VSTs following intravenous administration. We found that deletion of Itgb1, encoding for integrin beta 1, prevented OT-I cells from entering infected organs and drastically reduced their number in blood, suggesting that adoptively transferred VSTs primarily expand in the infected tissues. By contrast, Cxcr3-/- OT-I cells provided equal protection as their Cxcr3+/+ counterparts, indicating that this chemokine receptor does not contribute to VST entry into infected organs. Further, Pdcd1 and Ctla4 deletion did not impair the transferred OT-I cells' ability to protect mice from MCMV, arguing against quick exhaustion of VSTs with an effector T cell phenotype. Together, these data indicate that ex vivo expansion affects migration and activation properties of VSTs and suggest that future clinical evaluation of adoptive T cell therapy efficacy should include homing molecule expression assessment.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":"117 4","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144017630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Trained immunity enhances host resistance to infection in aged mice. 训练免疫增强老年小鼠宿主对感染的抵抗力。

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiae259

Dan Hao, Katherine R Caja, Margaret A McBride, Allison M Owen, Julia K Bohannon, Antonio Hernandez, Sabah Ali, Sujata Dalal, David L Williams, Edward R Sherwood

{"title":"Trained immunity enhances host resistance to infection in aged mice.","authors":"Dan Hao, Katherine R Caja, Margaret A McBride, Allison M Owen, Julia K Bohannon, Antonio Hernandez, Sabah Ali, Sujata Dalal, David L Williams, Edward R Sherwood","doi":"10.1093/jleuko/qiae259","DOIUrl":"10.1093/jleuko/qiae259","url":null,"abstract":"Aging significantly increases the incidence and severity of infections, with individuals aged 65 and above accounting for 65% of sepsis cases. Innate immune training, known as \"trained immunity\" or \"innate immune memory,\" has emerged as a potential strategy to enhance infection resistance by modulating the aging immune system. We investigated the impact of -glucan-induced trained immunity on aged mice (18 to 20 mo old) compared with young adult mice (10 to 12 wk old). Our findings showed that β-glucan equally augmented the host resistance to infection in both young and aged mice. This enhancement was characterized by augmented bacterial clearance, enhanced leukocyte β, and decreased cytokine production in response to Pseudomonas aeruginosa infection. Furthermore, young and aged trained macrophages displayed heightened metabolic capacity and improved antimicrobial functions, including enhanced phagocytosis and respiratory burst. RNA-seq analysis showed a distinctive gene expression pattern induced by trained immunity in macrophages characterized by activation of pathways regulating inflammation and the host response to infection and suppression of pathways regulating cell division, which was consistently observed in both young and aged groups. As compared with macrophages from young mice, aged macrophages showed increased activation of gene ontology pathways regulating angiogenesis, connective tissue deposition, and wound healing. Our results indicate that immune training can be effectively induced in aging mice, providing valuable insights into potential strategies for enhancing infection resistance in the elderly.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12022634/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142895300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Protocols for circulating neutrophil depletion in neonatal C57Bl/6 mice. 新生儿C57Bl/6小鼠循环中性粒细胞耗竭方案。

IF 3.6 3区医学

Journal of Leukocyte Biology Pub Date : 2025-04-23 DOI: 10.1093/jleuko/qiaf005

Devashis Mukherjee, Sriram Satyavolu, Sarah Cioffi, Asha Thomas, Yuexin Li, Lalitha Nayak

{"title":"Protocols for circulating neutrophil depletion in neonatal C57Bl/6 mice.","authors":"Devashis Mukherjee, Sriram Satyavolu, Sarah Cioffi, Asha Thomas, Yuexin Li, Lalitha Nayak","doi":"10.1093/jleuko/qiaf005","DOIUrl":"10.1093/jleuko/qiaf005","url":null,"abstract":"Murine neonatal neutrophil depletion strategies have problems achieving deep neutrophil clearance and accurate residual neutrophil fraction detection. An isotype switch method can achieve profound neutrophil clearance using a combination of anti-Ly6G and antirat κ Ig light chain antibodies in adult C57Bl/6 mice, proven by extra- and intracellular Ly6G detection by flow cytometry. We adapted this technique to neonatal mice, testing four neutrophil depletion strategies in the peripheral circulation, bone marrow, and spleen. Four protocols were tested: P3 Ly6G and P1-3 Ly6G (anti-Ly6G on postnatal days [P] 3 and 1-3, respectively), and P3 Dual and P1-3 Dual (anti-Ly6G and antirat κ Ig light chain on P3 and P1-3, respectively). Intracellular and extracellular Ly6G presence was detected using flow cytometry. Isotype control antibodies were used as controls. P1-3 Dual protocol achieved significantly better neutrophil depletion than the P1-3 Ly6G or P3 Ly6G protocols (97% vs 74% and 97% vs 50%, respectively) in the peripheral circulation. The P3 Dual protocol alone was enough to achieve significantly better neutrophil clearance (93%) than any of the Ly6G alone protocols. The Ly6G alone protocols led to near-total elimination of extracellular Ly6G. However, there was a significant presence of intracellular Ly6G in the CD45+ cell population, evading detection by extracellular Ly6G antibody-based detection methods. P3 protocols perform better than P1-3 protocols for bone marrow and splenic neutrophil clearance. Thus, the P3 Dual protocol might be the most effective and ethical protocol to induce profound neutrophil depletion in neonatal mice, an alternative to daily anti-Ly6G injections.","PeriodicalId":16186,"journal":{"name":"Journal of Leukocyte Biology","volume":" ","pages":""},"PeriodicalIF":3.6,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143006676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0