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OVA-8

OVA-8 is a protein that plays a key role in regulating cellular processes.
It is expressed in various tissues and has been implicated in a number of physiological and pathological conditions.
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Most cited protocols related to «OVA-8»

1
T Cell Dynamics in Lymph Node
Cited 10 times
Nine time lapse videos illustrate many of the key findings. Video S1 illustrates the kinetics of T cell homing to a draining inguinal lymph node. Video S2 shows stochastic T cell repertoire scanning in the absence of OVA. Video S3 provides a side-by-side comparison of T cell motility in the presence and absence of antigen. Video S4 depicts early cognate interactions (1–3 h) between antigen-specific T cells and DCs in the draining lymph node of OVA-challenged mice. Video S5 shows T cells forming dynamic clusters on DC dendrites at later times (8 h) in OVA-challenged mice. Video S6 shows that clusters are less apparent in experiments with reduced amounts of OVA. Video S7 shows that OVA-specific T cells regain motility and move in “swarms” near DCs (16 h). Video S8 shows reduced T cell–DC interaction 24 h after adoptive transfer. Video S9 depicts T cell motility and proliferation 26 h after adoptive transfer. Online supplemental material is available at http://www.jem.org/cgi/content/full/jem.20041236/DC1.
Miller M.J., Safrina O., Parker I, & Cahalan M.D. (2004). Imaging the Single Cell Dynamics of CD4+ T Cell Activation by Dendritic Cells in Lymph Nodes. The Journal of Experimental Medicine, 200(7), 847-856.
Publication 2004
Adoptive Cell Transfer Adoptive Transfer Antigens Dendrites Groin H antigen, bacterial Kinetics Motility, Cell Mus Nodes, Lymph OVA-8 T-Lymphocyte
2
Detecting Antigen-Specific CD8+ T Cells
Cited 9 times
The following antibodies were purchased from BD Biosciences: anti–mouse CD8-APC (Ly-2), anti–mouse CD44-FITC (Pgp-1, Ly-24), and B220-Cy (RA3–6B2). CD40 (1C10 or FGK45) were produced by hybridomas that were grown in serum-free conditions. Ova-specific CD8 T cells were detected by H-2Kb–specific tetramers containing the SIINFEKL peptide, either made as described previously (33 (link)) (a gift from Dr. Lefrancois, University of Connecticut Health Center, Farmington, CT) or purchased from Beckman Coulter.
Ahonen C.L., Doxsee C.L., McGurran S.M., Riter T.R., Wade W.F., Barth R.J., Vasilakos J.P., Noelle R.J, & Kedl R.M. (2004). Combined TLR and CD40 Triggering Induces Potent CD8+ T Cell Expansion with Variable Dependence on Type I IFN. The Journal of Experimental Medicine, 199(6), 775-784.
Publication 2004
Antibodies CD8-Positive T-Lymphocytes CD44 protein, human FGK45 monoclonal antibody Fluorescein-5-isothiocyanate Hybridomas Mus OVA-8 Serum Tetrameres
3
Discriminating Tissue-Resident CD8 T Cells
Cited 7 times
To discriminate between CD8 T cells in tissue parenchyma versus tissue vasculature, i.v. injected Ab was used as previously described (28 (link)). Briefly, 3μg of anti-CD8α Ab (53-6.7, Biolegend, San Diego, CA) was injected i.v. and allowed to circulate for three minutes prior to mouse sacrifice.
Organs were harvested and digested as previously described (29 (link)). For isolation of small intestinal intraepithelial lymphocytes (IEL), Peyer’s patches were removed, the small intestine was cut longitudinally and then laterally into small pieces. Pieces were incubated for 30 minutes with stirring at 37°C with 0.154mg/mL dithioerythritol (Sigma-Aldrich, St. Louis, MO) in 10% HBSS/HEPES. Female reproductive tract (FRT), lung and salivary gland (SG) tissues were cut into small pieces in RPMI 1640 containing 5% FBS, 2 mM MgCl2, 2 mM CaCl2 and 0.5mg/mL type IV collagenase for FRT (Sigma-Aldrich, St. Louis, MO) or 100 U/mL type I collagenase for lung and SG (Worthington, Lakewood, NJ) and incubated for 1 hr at 37°C with stirring. After enzymatic digestion, the remaining tissue pieces were mechanically disrupted using a gentleMACs dissociator (Miltenyi Biotec, San Diego, CA). The liver was mechanically dissociated by pushing the tissue through a 70μm-cell strainer. Single cell suspensions of IEL, FRT, lung, liver and SG were further separated using a 44/67% Percoll (GE Healthcare Life Sciences, Pittsburgh, PA) density gradient. Spleen and lymph nodes (LN) were dissociated mechanically. Splenocytes and blood were treated with ACK lysis buffer to lyse red blood cells.
The following antibodies were used for flow cytometry: anti-KLRG1 (2F1), anti-Eomes (Dan11ma), anti-T-bet (4B10), anti-CD44 (IM7), anti-CD122 (TM-b1), anti-CD27 (LG.7F9), anti-CD69 (H1.2F3) (all from eBioscience, San Diego, CA), anti-CD8α (53-6.7, eBioscience, Biolegend, San Diego, CA), anti-CD103 (M290), anti-CD25 (PC61), anti-Bcl-2 (Bcl-2/100) and anti-CD127 (SB/199) (BD Biosciences, San Jose, CA). Cell viability was determined using Ghost DyeTM Red 780 (Tonbo Biosciences, San Diego, CA). Kb-SIINFEKL-specific CD8 T cells were identified using H-2Kb tetramers made in house containing the SIINFEKL peptide (New England Peptide, Gardener, MA). The BD Biosciences intracellular kit for cytokine staining and the eBioscience FoxP3 kit for transcription factor staining were used in accordance with manufacturer’s directions. Peptide stimulation was performed as previously described (30 (link)). Briefly, splenocytes were plated in RPMI 1640 containing 10% FBS, 1× NEAA, 2mM L-glutamine, 1mM sodium pyruvate, 1× penicillin/streptomycin and 0.05mM β-mercaptoethanol and incubated with 1ug/mL SIINFEKL peptide and 1ug/mL GolgiPlug (BD Biosciences, San Jose, CA) for four hours at 37°C. Cells were washed and stained with fixable LIVE/DEAD aqua dead cell stain (Life Technologies, San Diego, CA) before surface and intracellular staining. Samples were acquired on an LSRII flow cytometer (BD Biosciences, San Diego, CA).
Thompson E.A., Beura L.K., Nelson C.E., Anderson K.G, & Vezys V. (2016). Shortened intervals during heterologous boosting preserve memory CD8 T cell function but compromise longevity. Journal of immunology (Baltimore, Md. : 1950), 196(7), 3054-3063.
Publication 2016
2-Mercaptoethanol alpha HML-1 Antibodies BCL2 protein, human BLOOD Buffers CD8-Positive T-Lymphocytes CD44 protein, human Cells Cell Survival Collagenase, Clostridium histolyticum Cytokine Digestion Dithioerythritol Enzymes Erythrocytes Female Reproductive System Flow Cytometry Glutamine Hemoglobin, Sickle HEPES IL2RA protein, human IL2RB protein, human Intestines, Small Intraepithelial Lymphocytes isolation KB Cells KLRG1 protein, human Liver Lung Lymphocyte Magnesium Chloride Matrix Metalloproteinase 2 Mus Nodes, Lymph OVA-8 Penicillins Peptides Percoll Peyer Patches Protoplasm Pyruvate Red Cell Ghost Salivary Glands Sodium Spleen Stains Streptomycin Tetrameres Thomsen-Friedenreich antibodies Tissues Transcription Factor
4
Murine Model of Allergic Asthma
Cited 6 times
Mice in the naïve control group received no treatment for the duration of the experiment. Mice in the inhaled saline group were sensitized intraperitoneally with 200 μl saline on days 1 and 8. On days 15, 16, and 17 after initial sensitization with saline, mice were nebulized with saline for 30 min in a whole-body exposure chamber. Mice in the intranasal saline group were intraperitoneally sensitized as inhaled saline group and then intranasally instilled with 40 μl saline on days 15, 16, and 17. Mice in the inhaled OVA group were intraperitoneally sensitized on days 1 and 8 with 20 μg OVA (Sigma-Aldrich, St. Louis, MO) emulsified in 1 mg aluminum hydroxide (Thermo Fisher Scientific, Waltham, MA) in a total volume of 200 μl. On days 15, 16, and 17 after initial sensitization with OVA, mice were administered aerosolized 5% (v/v) OVA by inhalation for 30 min in a whole-body exposure chamber. Mice in the intranasal OVA group were intraperitoneally sensitized as the inhaled OVA group and then intranasally instilled with 20 μg OVA on days 15, 16, and 17. For intranasal instillation, mice were anesthetized using inhaled anesthesia. Prior to instillation, isoflurane was delivered into induction chamber using small animal portable anesthesia systems (L-PAS-02, LMSKOREA, Inc., Seongnam, Korea) equipped with isoflurane vaporizer. And then, mice were exposed to 2.5% isoflurane delivered in O2 (2 L/min) within induction chamber until a sleep-like state was reached. Mice receiving isoflurane anesthesia were removed from the induction chamber and instillation was performed immediately on board. Intranasal administration of challenge dose (saline or 20 μg OVA diluted in 40 μl saline) was performed by pipetting onto the outer edge of nose of the mice. After instillation, mice moved, fully recovered and transferred to their cage.
Twenty-four hours after the last challenge with OVA, AHR were assessed. And 48 h after the last challenge with OVA, mice were euthanized with an overdose of isoflurane and continuously exposed until 1 min after breathing stops. The sample collections for analysis were performed in sacrificed animals. Lung inflammation, serum IgE production, and protein levels of inflammatory and epithelial cell-derived cytokines were assessed.
Kim D.I., Song M.K, & Lee K. (2019). Comparison of asthma phenotypes in OVA-induced mice challenged via inhaled and intranasal routes. BMC Pulmonary Medicine, 19, 241.
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Publication 2019
Administration, Intranasal Anesthesia Animals Apnea Cytokine Drug Overdose Epithelial Cells External Nose Human Body Hydroxide, Aluminum Inflammation Isoflurane Mus OVA-8 Pneumonia Proteins Saline Solution Serum Sleep Specimen Collection Vaporizers
5
Antigen-Specific T Cell Activation
Cited 6 times
6–12-wk-old female C57BL/6J mice were injected i.p., unless otherwise noted, with 0.5 mg whole ovalbumin (Sigma-Aldrich) with or without varying amounts of TLR agonists and/or 50 μg of 1C10 or FGK45 (anti-CD40). Where peptide injections are noted, mice were injected with anti-CD40 i.p. and then 4–6 h later injected i.v. with 100 mg SIINFEKL peptide and a given TLR agonist. Anti-CD40 was used at 50 μg per injection. Mice were immunized with a single injection i.p. and killed 6 d later unless otherwise noted.
Ahonen C.L., Doxsee C.L., McGurran S.M., Riter T.R., Wade W.F., Barth R.J., Vasilakos J.P., Noelle R.J, & Kedl R.M. (2004). Combined TLR and CD40 Triggering Induces Potent CD8+ T Cell Expansion with Variable Dependence on Type I IFN. The Journal of Experimental Medicine, 199(6), 775-784.
Publication 2004
agonists Females FGK45 monoclonal antibody Mice, House Mice, Inbred C57BL OVA-8 Ovalbumin Peptides

Most recents protocols related to «OVA-8»

1
SIINFEKL-specific CD8+ T Cell Assay
The liver cells were incubated with Fc-blocking reagent (TrueStain, Biolegend) for 5 min, incubated with APC-SIINFEKL-H-2Kb dextramers (Immunotools) for 10 min and then stained with the appropriate antibodies detailed in the supplementary files. For the assessment of the MHC class I molecule Kb bound to the ovalbumin peptide SIINFEKL, RIL-175-LV-OVA-GFP or RIL-175-LV-GFP cells were left untreated or incubated overnight with 250 ng/ml γIFN (Peprotec) and then labeled with the APC anti-SIINFEKL-H2Kb antibody (clone 25-D1.16). The samples were processed with an Attune NxT Cytometer (ThermoScientific) and analyzed with FloJo (Treestar). Results of FACS analysis are presented as the percentage of a population or as mean fluorescence index (MFI).
Lacotte S., Slits F., Moeckli B., Peloso A., Koenig S., Tihy M., El Hajji S., Gex Q., Rubbia-Brandt L, & Toso C. (2023). Anti-CD122 antibody restores specific CD8+ T cell response in nonalcoholic steatohepatitis and prevents hepatocellular carcinoma growth. Oncoimmunology, 12(1), 2184991.
Publication 2023
Antibodies Antibodies, Anti-Idiotypic Cells Clone Cells Fluorescence Hepatocyte Histocompatibility Antigens Class I OVA-8 Ovalbumin
2
SIINFEKL-Pulsed BMDCs Immunization
BMDCs from C57BL/6N WT were pulsed with SIINFEKL peptide (0.5 µg/106 cells/1 mL) together with LPS (1 µg/mL) for 2 h at 37 °C. Control cells were pulsed with LPS alone at the same concentration. The cells were resuspended in PBS prior i.v. injections to WT and HBB mice (1 × 106 cells/mice) on days 0 and 7. On day 10, blood samples (100 µL) were collected from the facial vein and PBL analyzed by FACS (SI Appendix). On day 12, all mice were killed by cervical dislocation and lymph nodes and spleens collected. Single-cell suspensions of 5 × 106 cells were prepared for FACS analysis (SI Appendix).
Sroka E.M., Lavigne M., Pla M., Daskalogianni C., Tovar-Fernandez M.C., Prado Martins R., Manoury B., Darrasse-Jéze G., Nascimento M., Apcher S, & Fåhraeus R. (2023). Major histocompatibility class I antigenic peptides derived from translation of pre-mRNAs generate immune tolerance. Proceedings of the National Academy of Sciences of the United States of America, 120(7), e2208509120.
Publication 2023
BLOOD Cells Face Joint Dislocations Mus Neck Nodes, Lymph OVA-8 Veins
3
Splenocyte Activation by SIINFEKL and TGF-β
Splenocytes were cultured in vitro for 24 hours in 1nm SIINFEKL peptide or with 10ng/mL of TGF-β in RPMI containing 10% FBS.
Heim T.A., Lin Z., Steele M.M., Mudianto T, & Lund A.W. (2023). CXCR6 promotes dermal CD8+ T cell survival and transition to long-term tissue residence. bioRxiv.
Publication Preprint 2023
OVA-8 Transforming Growth Factor beta
4
Oral Tolerance Induction and OVA-Specific Immune Responses
Oral tolerance was induced by continuously feeding animals with 8 mg/mL of OVA in the drinking water for 5 days. Control mice received only water. Three days after the last feeding, mice were immunized with 50 µg of OVA in complete Freund’s adjuvant (CFA) in the ventral flanks. In vitro recall responses were measured at day 10 after immunization. For this, splenocytes were stimulated with 4, 20, and 100 µg/mL of LPS-free OVA (antigen-specific stimulation) or 1 µg/mL of anti-CD3 (antigen non-specific stimulation as a control for proliferation), and proliferation was measured using 3H-thymidine incorporation. In some experiments, tolerance to OVA was measured by delayed type hypersensitivity (DTH) by injecting 60 µg of OVA into the hind paw and measuring foot pad swelling 1, 2, and 3 days later.
Rezende R.M., Cox L.M., Moreira T.G., Liu S., Boulenouar S., Dhang F., LeServe D.S., Nakagaki B.N., Lopes J.R., Tatematsu B.K., Lemos L., Mayrink J., Lobo E.L., Guo L., Oliveira M.G., Kuhn C, & Weiner H.L. (2023). Gamma-delta T cells modulate the microbiota and fecal micro-RNAs to maintain mucosal tolerance. Microbiome, 11, 32.
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Publication 2023
Antigens Delayed Hypersensitivity Foot Freund's Adjuvant Immune Tolerance Mental Recall Muromonab-CD3 Mus OVA-8 Thymidine Vaccination
5
OT-1 Cell-Mediated Cytotoxicity Assay
Splenocytes (106/mL) from OT1 mice were stimulated in RPMI + 10% FCS with 1mM OVA peptide for 3 days, washed and cultured with IL2 as above for 3 more days. Targets EL-4 cells loaded with Cell Trace Violet (C34557) at 2uM in PBS for 10 minutes then quenched and washed. Cells were pulsed with 1uM SIINFEKL peptide for 30-60 minutes at 37 degrees. Target cells were incubated with activated OT-1 cells, at varying ratios (1:5, 1:10, 1:20 in 200 mL media for 4 hours. Stained w/APC-anti-CD8 and live/dead for 30 minutes on ice. Acquired on Beckman CytoFLEX cytometer.
Tarasenko T.N., Banerjee P., Gomez-Rodriguez J., Gildea D., Zhang S., Wolfsberg T., Jenkins L.M, & McGuire P.J. (2023). Pyruvate dehydrogenase complex integrates the metabolome and epigenome in memory T cell differentiation in vitro. Research Square.
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Publication Preprint 2023
Cells Mus OVA-8 Peptides Viola

Top products related to «OVA-8»

1
Siinfekl peptide by AnaSpec
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SIINFEKL peptide is a synthetic peptide sequence commonly used in research applications. It is a short amino acid sequence that can be utilized in various experimental settings. The core function of this product is to serve as a tool for researchers, without further interpretation or extrapolation on its intended use.
2
Siinfekl peptide by Merck Group
Sourced in United States
SIINFEKL peptide is a synthetic peptide used in research applications. It is a commonly used antigen for studying immune responses in mice. The SIINFEKL peptide is derived from the chicken ovalbumin protein and is presented by the H-2Kb class I major histocompatibility complex (MHC) molecule.
3
Carboxyfluorescein succinimidyl ester (cfse) by Thermo Fisher Scientific
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CFSE (Carboxyfluorescein succinimidyl ester) is a fluorescent dye used for cell proliferation and tracking assays. It binds to cellular proteins, allowing the labeling and monitoring of cell division in various cell types.
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GolgiPlug is a laboratory product designed to inhibit protein transport from the Golgi apparatus to the cell surface. It functions by blocking the secretory pathway, preventing the release of proteins from the Golgi complex. GolgiPlug is intended for use in cell biology research applications.
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Brefeldin A is a fungal metabolite that inhibits the function of Golgi apparatus in eukaryotic cells. It acts by blocking the exchange of materials between the endoplasmic reticulum and Golgi compartments, leading to the collapse of the Golgi structure.
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SIINFEKL peptide is a synthetic peptide with the sequence SIINFEKL. It is commonly used as a model antigen in immunological studies.
7
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SIINFEKL peptide is a synthetic peptide that consists of the amino acid sequence SIINFEKL. This peptide is commonly used in immunological research to study antigen presentation and T cell activation.
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Cytofix cytoperm kit by BD
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The Cytofix/Cytoperm kit is a laboratory product designed for fixing and permeabilizing cells. It provides the necessary solutions for the preparation of samples prior to intracellular staining and flow cytometric analysis.
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IL-2 is a cytokine that plays a crucial role in the regulation of the immune system. It is a protein produced by T-cells and natural killer cells, and it is essential for the activation, proliferation, and differentiation of these cells. IL-2 is an important component in various immunological processes, including the promotion of T-cell growth and the enhancement of the cytolytic activity of natural killer cells.

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