Evaluation of the relative expression of genes associated with adherence after different hours of co-culture between Streptococcus uberis and MAC-T cells

Abstract Conceptus secretory factors include galectins, a family of carbohydrate binding proteins that elicit cell adhesion and immune suppression by interacting with intracellular and extracellular glycans. In rodents, galectin-1 (LGALS1) promotes maternal-fetal immune tolerance in the decidua through expansion of tolerogenic CD11c+ dendritic cells, increased anti-inflammatory IL-10, and activation of FOXP3+ regulatory T cells (Treg). This study characterized galectin expression in early ruminant conceptuses and endometrium. We also tested the effect of recombinant bovine LGALS1 (rbLGALS1) and progesterone (P4) on endometrial expression of genes and protein related to maternal-fetal immune tolerance in cattle. Elongating bovine and ovine conceptuses expressed several galectins, particularly, LGALS1, LGALS3 and LGALS8. Within bovine endometrium, expression of LGALS3, LGALS7 and LGALS9 was greater on Day 16 of pregnancy compared to the estrous cycle. Within ovine endometrium, LGALS7 was greater during pregnancy compared to the estrous cycle and endometrium of pregnant sheep tended to have greater LGALS9 and LGALS15. Expression of endometrial LGALS4 was less during pregnancy in sheep. Treating bovine endometrium with rbLGALS1 increased endometrial expression of CD11c, IL-10 and FOXP3, within 24 h. Specifically, within caruncular endometrium, both rbLGALS1 and P4 increased FOXP3, suggesting that both ligands may promote Treg expansion. Using IHC, FOXP3+ cells with a leukocyte phenotype were localized to the bovine uterine stratum compactum near the uterine surface and increased in response to rbLGALS1. We hypothesize that galectins have important functions during establishment of pregnancy in ruminants and bovine conceptus LGALS1 and luteal P4 confer mechanisms of maternal-conceptus immune tolerance in cattle.

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Relative expression of mRNAs related to cavitation process in bovine embryos produced in vivo and in vitro

Revista Brasileira de Zootecnia ◽

10.1590/s1516-35982011000100017 ◽

2011 ◽

Vol 40 (1) ◽

pp. 124-128

Author(s):

Sabine Wohlres-Viana ◽

Mariana Cortes Boite ◽

João Henrique Moreira Viana ◽

Marco Antonio Machado ◽

Luiz Sérgio de Almeida Camargo

Keyword(s):

Culture System ◽

Rna Extraction ◽

Endogenous Control ◽

Bovine Embryos ◽

Relative Expression ◽

Gapdh Gene ◽

Expression Of Genes ◽

In Vitro Culture System

The objectives of this work were to identify and to evaluate possible differences on gene expression of aquaporins and Na/K-ATPases transcripts between embryos in vivo and in vitro produced. For each group, 15 blastocysts distributed in three pools were used for RNA extraction followed by amplification and reverse transcription. The resulting cDNAs were submitted to Real-Time PCR, using the GAPDH gene as endogenous control. It was not possible to identify AQP1 transcripts. Relative expression of AQP3 (1.33 ± 0.78) and AQP11 (2.00 ± 1.42) were not different in blastocysts in vitro and in vivo produced. Na/K-ATPase α1 gene (2.25 ± 1.07) was overregulated whereas Na/K-ATPase β2 transcripts 0.40 ± 0.30) did not differ among blastocysts produced in vitro from those produced in vivo. Transcripts for gene AQP1 are not present in bovine blastocysts. In vitro culture system does not alter expression of genes AQP3, AQP11 and Na/K-ATPase β2 genes, however, it affects expression of Na/K-ATPase α1.

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Prioritization of autoimmune disease-associated genetic variants that perturb regulatory element activity in T cells

10.1101/2021.05.30.445673 ◽

2021 ◽

Author(s):

Kousuke Mouri ◽

Michael H. Guo ◽

Carl G. de Boer ◽

Greg A. Newby ◽

Matteo Gentili ◽

...

Keyword(s):

T Cells ◽

Autoimmune Disease ◽

Association Studies ◽

Regulatory Element ◽

Genome Wide Association Studies ◽

Expression Of Genes ◽

Human T Cell ◽

Allele Specific ◽

Reporter Assays ◽

Element Activity

Genome-wide association studies have uncovered hundreds of autoimmune disease-associated loci; however, the causal genetic variant(s) within each locus are mostly unknown. Here, we perform high-throughput allele-specific reporter assays to prioritize disease-associated variants for five autoimmune diseases. By examining variants that both promote allele-specific reporter expression and are located in accessible chromatin, we identify 60 putatively causal variants that enrich for statistically fine-mapped variants by up to 57.8-fold. We introduced the risk allele of a prioritized variant (rs72928038) into a human T cell line and deleted the orthologous sequence in mice, both resulting in reduced BACH2 expression. Naive CD8 T cells from mice containing the deletion had reduced expression of genes that suppress activation and maintain stemness. Our results represent an example of an effective approach for prioritizing variants and studying their physiologically relevant effects.

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Characterization of human FDCs reveals regulation of T cells and antigen presentation to B cells

Journal of Experimental Medicine ◽

10.1084/jem.20210790 ◽

2021 ◽

Vol 218 (10) ◽

Cited By ~ 1

Author(s):

Balthasar A. Heesters ◽

Kyah van Megesen ◽

Ilhan Tomris ◽

Robert P. de Vries ◽

Giuliana Magri ◽

...

Keyword(s):

T Cells ◽

B Cells ◽

B Cell ◽

Intimate Contact ◽

Expression Of Genes ◽

T Cell Regulation ◽

Native Antigen ◽

Reticular Cells ◽

Molecular Patterns

Stromal-derived follicular dendritic cells (FDCs) are essential for germinal centers (GCs), the site where B cells maturate their antibodies. FDCs present native antigen to B cells and maintain a CXCL13 gradient to form the B cell follicle. Yet despite their essential role, the transcriptome of human FDCs remains undefined. Using single-cell RNA sequencing and microarray, we provided the transcriptome of these enigmatic cells as a comprehensive resource. Key genes were validated by flow cytometry and microscopy. Surprisingly, marginal reticular cells (MRCs) rather than FDCs expressed B cell activating factor (BAFF). Furthermore, we found that human FDCs expressed TLR4 and can alter antigen availability in response to pathogen-associated molecular patterns (PAMPs). High expression of PD-L1 and PD-L2 on FDCs activated PD1 on T cells. In addition, we found expression of genes related to T cell regulation, such as HLA-DRA, CD40, and others. These data suggest intimate contact between human FDCs and T cells.

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Naïve Regulatory T Cell Subset Is Altered in X-Linked Agammaglobulinemia

Frontiers in Immunology ◽

10.3389/fimmu.2021.697307 ◽

2021 ◽

Vol 12 ◽

Author(s):

Pavel V. Shelyakin ◽

Ksenia R. Lupyr ◽

Evgeny S. Egorov ◽

Ilya A. Kofiadi ◽

Dmitriy B. Staroverov ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

B Cell ◽

Cell Subset ◽

Cell Receptor ◽

T Cell Subsets ◽

Cell Compartments ◽

Expression Of Genes ◽

Tcr Repertoire ◽

Antigen Presenting

The interplay between T- and B-cell compartments during naïve, effector and memory T cell maturation is critical for a balanced immune response. Primary B-cell immunodeficiency arising from X-linked agammaglobulinemia (XLA) offers a model to explore B cell impact on T cell subsets, starting from the thymic selection. Here we investigated characteristics of naïve and effector T cell subsets in XLA patients, revealing prominent alterations in the corresponding T-cell receptor (TCR) repertoires. We observed immunosenescence in terms of decreased diversity of naïve CD4+ and CD8+ TCR repertoires in XLA donors. The most substantial alterations were found within naïve CD4+ subsets, and we have investigated these in greater detail. In particular, increased clonality and convergence, along with shorter CDR3 regions, suggested narrower focused antigen-specific maturation of thymus-derived naïve Treg (CD4+CD45RA+CD27+CD25+) in the absence of B cells - normally presenting diverse self and commensal antigens. The naïve Treg proportion among naïve CD4 T cells was decreased in XLA patients, supporting the concept of impaired thymic naïve Treg selection. Furthermore, the naïve Treg subset showed prominent differences at the transcriptome level, including increased expression of genes specific for antigen-presenting and myeloid cells. Altogether, our findings suggest active B cell involvement in CD4 T cell subsets maturation, including B cell-dependent expansion of the naïve Treg TCR repertoire that enables better control of self-reactive T cells.

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Primary vulvar squamous cell carcinomas with high T cell infiltration and active immune signaling are potential candidates for neoadjuvant PD-1/PD-L1 immunotherapy

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2021-003671 ◽

2021 ◽

Vol 9 (10) ◽

pp. e003671

Author(s):

Kim E Kortekaas ◽

Saskia J Santegoets ◽

Liselotte Tas ◽

Ilina Ehsan ◽

Pornpimol Charoentong ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Signaling Pathways ◽

T Cell Activation ◽

Squamous Cell ◽

Molecular Subtype ◽

Cell Infiltration ◽

Immune Signaling ◽

T Cell Infiltration ◽

Expression Of Genes

BackgroundA profound insight into the immune landscape of vulvar squamous cell carcinoma (VSCC) is lacking. Here, an in-depth interrogation of T cell infiltration, local immune contexture, signaling pathways and checkpoint molecule expression was performed in early-stage and late-stage VSCC.MethodsThe type, location, and infiltration pattern of T cells were studied in 109 patients with primary VSCC FIGO stage I–III. RNA expression of genes involved in immune oncology and oncogenic signaling pathways was analyzed in 40 VSCC, matched for prognostic clinicopathological variables, analyzed for HPV and p53 status, and selected based on T cell infiltration.ResultsHigh intraepithelial infiltration with CD4 or CD8 T cells was associated with longer overall and recurrence-free survival and formed an independent prognostic factor, outperforming molecular subtype and stage of the disease. Strong T cell infiltrated VSCC displayed a coordinated immune response reflected by a positive association between T cells and different lymphocyte and myeloid cell subsets. The expression of genes involved in the migration of T cells and myeloid cells, T cell activation and costimulation, interferon (IFN)-γ signaling, cytotoxicity and apoptosis was higher than in low infiltrated tumors. An active immune signaling profile was observed in all inflamed, part of the altered-excluded and not in altered-immunosuppressed or deserted VSCC. While several checkpoint molecules were overexpressed, only PD-L1 expression displayed discriminatory ability and clinical usefulness. High PD-L1 expression was detected in all inflamed and ~60% of the altered-excluded VSCC.ConclusionAn active immune signaling profile is present in 35% of primary FIGO I–III VSCCs, suggesting potential responsiveness to neoadjuvant PD-1/PD-L1 immunotherapy.

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T lymphocytes potentiate endogenous neuroprotective inflammation in a mouse model of ALS

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.0804610105 ◽

2008 ◽

Vol 105 (46) ◽

pp. 17913-17918 ◽

Cited By ~ 209

Author(s):

Isaac M. Chiu ◽

Adam Chen ◽

Yi Zheng ◽

Bela Kosaras ◽

Stefanos A. Tsiftsoglou ◽

...

Keyword(s):

T Cells ◽

Mouse Model ◽

Disease Progression ◽

Cholesterol Metabolism ◽

Cell Receptor ◽

Specific Flow ◽

Innate And Adaptive Immunity ◽

Expression Of Genes ◽

Lateral Sclerosis

Amyotrophic Lateral Sclerosis (ALS) is an adult-onset, progressive, motor neuron degenerative disease, in which the role of inflammation is not well established. Innate and adaptive immunity were investigated in the CNS of the Superoxide Dismutase 1 (SOD1)G93A transgenic mouse model of ALS. CD4+ and CD8+ T cells infiltrated SOD1G93A spinal cords during disease progression. Cell-specific flow cytometry and gene expression profiling showed significant phenotypic changes in microglia, including dendritic cell receptor acquisition, and expression of genes linked to neuroprotection, cholesterol metabolism and tissue remodeling. Microglia dramatically up-regulated IGF-1 and down-regulated IL-6 expression. When mutant SOD1 mice were bred onto a TCRβ deficient background, disease progression was significantly accelerated at the symptomatic stage. In addition, microglia reactivity and IGF-1 levels were reduced in spinal cords of SOD1G93A (TCRβ−/−) mice. These results indicate that T cells play an endogenous neuroprotective role in ALS by modulating a beneficial inflammatory response to neuronal injury.

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Investigation the Relative Expression of Genes of ATPase Family in Major and Regular Depressed Patients

International Journal of Indian Psychology ◽

10.25215/0304.014 ◽

2016 ◽

Vol 3 (4) ◽

Author(s):

Sima Farzaneh ◽

Tavkol Moussazadeh ◽

Mohammad Zaefizadeh

Keyword(s):

Quantitative Measurement ◽

Comparison Group ◽

Rt Pcr ◽

Statistical Population ◽

Relative Expression ◽

Depressed Patients ◽

Normal People ◽

Expression Of Genes ◽

Significant Difference ◽

Comprehensive Manner

To develop new pharmaceutical strategies in a comprehensive manner, it is necessary to understand the reasons for major depression. Present paper aims for comparison of expression of ATPase families in major depressed and normal people. Statistical population of this research includes all depressed people coming to professional clinic of Shahryar in 2014-15. This contains 47 depressed patients which were selected based on accessibility from those visiting clinic. 26 normal people were selected as comparison group as well. To study the relative expression of ATPase1-3, quantitative measurement method using Real-Time RT-PCR was used. Relative expression of ATPase2 showed a significant reduction compared to normal people (p<0.02). In patients, ATPase1 had higher relative expression compared to normal people. However, ATPase had no significant difference among patients and normal people. Results demonstrate that ATPase1 and ATPase2 are important variables in depression but ATPase3 plays no pivotal role in this regard.

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Basal anti-inflammatory responses in cultured endothelial cells exposed to two conjugated linoleic acids

Proceedings of The Nutrition Society ◽

10.1017/s0029665120004310 ◽

2020 ◽

Vol 79 (OCE2) ◽

Author(s):

Carina Valenzuela ◽

Elizabeth Miles ◽

Philip Calder

Keyword(s):

Inflammatory Responses ◽

Adhesion Assay ◽

Dependent Manner ◽

Relative Expression ◽

Inflammatory Pathways ◽

Expression Of Genes ◽

Low Concentrations ◽

Genes Encoding ◽

Anti Inflammatory ◽

Pre Treatment

AbstractConjugated linoleic acid (CLA) isomers have been shown to possess anti-atherosclerotic properties, which may be related to the downregulation of inflammatory pathways. Whether low concentrations of CLAs are able to affect basal, unstimulated endothelial cell (EC) responses is not clear. The aim of this study was to evaluate the effects of two CLAs (cis-9, trans-11 and trans-10, cis-12) on basal inflammatory responses by ECs.EA.hy926 cells (HUVEC lineage) were cultured under standard conditions and exposed to CLAs (1 and 10 μM) for 48 hours. MTT assay was performed to determine cell viability; incorporation of FA was confirmed by gas chromatography; inflammatory mediators were assessed by multiplex immunoassay; the relative expression of genes encoding transcription factors and inflammatory cytokines was assessed through real-time PCR and static adhesion assay was used to evaluate monocyte attachment to the EC monolayer.CLAs were incorporated into ECs in a dose-dependent manner. Pre-treatment with CLA9,11 (1 uM) significantly reduced unstimulated, basal concentrations of MCP-1 (p < 0.05), and CLA10,12 at 10 uM had the same effect (p < 0.05). Both CLAs at 10 uM increased the relative expression of NFκβ (p < 0.01 and p < 0.05, respectively), while decreasing the relative expression of PPARα (p < 0.0001), COX-2 (p < 0.0001) and IL-6 (p < 0.0001). In contrast, no effect was observed in the adhesion assay for either CLA.These results suggest that both CLAs at a low concentration have a neutral or modest anti-inflammatory effect in basal conditions, which may influence endothelial function and risk of vascular disease. Interestingly, at these low CLA concentrations some pro-inflammatory genes are upregulated while others are down regulated. This suggests complex effects of CLAs on inflammatory pathways.

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