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Lipid a

Manufactured by Peptide Institute
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Lipid A is a key component of the outer membrane of gram-negative bacteria. It serves as the hydrophobic anchor of lipopolysaccharides, which are crucial structural elements of the bacterial cell wall. Lipid A plays a central role in the innate immune response and is an important target for research in microbiology, immunology, and drug development.

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Lab products found in correlation

Muramyl dipeptide mdp, by Bachem (4 mentions) Pam3cys, by Merck Group (4 mentions) Il 10, by BD (3 mentions) Il 1β, by Thermo Fisher Scientific (3 mentions) Gm csf, by Thermo Fisher Scientific (3 mentions) Cpg dna, by InvivoGen (3 mentions) Flagellin, by InvivoGen (2 mentions) Interleukin 4 (il 4), by R&D Systems (2 mentions) Tri dap, by InvivoGen (2 mentions) Il 23, by Thermo Fisher Scientific (2 mentions) Ebio473p19, by Thermo Fisher Scientific (1 mentions) Ifn γ, by Thermo Fisher Scientific (1 mentions) Fludarabine, by Cayman Chemical (1 mentions) Ifn α, by Abcam (1 mentions) Upadacitinib, by MedChemExpress (1 mentions) Il 12, by Thermo Fisher Scientific (1 mentions) Ifn γ, by BioLegend (1 mentions) Ifn γ, by R&D Systems (1 mentions) Ifn γ, by BD (1 mentions) Il 12, by R&D Systems (1 mentions)

Close spelling variants of this product

Alcaligenes lipid A (1 citations)

12 protocols using lipid a

1

Macrophage Immune Response Profiling

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Monocyte-derived macrophages (MDMs) were treated with muramyl dipeptide (MDP; Bachem, Torrance, CA), lipid A (Peptides International, Louisville, KY), Pam3Cys (EMD Millipore, Billerica, MA), polyI:C, flagellin, CL097, CpG (InvivoGen, San Diego, CA), IL23 (PeproTech), 10:1 multiplicity of infection Salmonella enterica serovar Typhimurium or adherent invasive Escherichia coli (AIEC) strain LF82. In some cases cells were incubated with neutralising anti-IL23p19 antibody (eBioscience, San Diego, CA) for 1 hour before the treatments. Supernatants were assayed for tumour necrosis factor (TNF), IL6, IL8, IL10 (BD Biosciences, San Jose, CA), IL1β or IL23 (coating antibody to IL23p19, clone eBio473P19, detection antibody to IL12/ IL23p40, clone C8.6) (eBioscience) by ELISA.
Sun R., Hedl M, & Abraham C. (2019). IL23 induces IL23R recycling and amplifies innate receptor-induced signalling and cytokines in human macrophages, and the IBD-protective IL23R R381Q variant modulates these outcomes. Gut, 69(2), 264-273.
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2

Generating Murine Bone Marrow-Derived Dendritic Cells and Macrophages

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BM single cell suspensions were cultured in complete RPMI 1640 media containing 20 ng/mL GM-CSF (PeproTech, Rocky Hill, NJ) (for BMDC) and 10% L929-conditioned medium (for BMM). Cultures were fed fresh medium every 3 days, and used at 6 to 8 days. Purity was >98% as assessed by flow cytometry. BMDC were CD11c+ and F4/80−, while BMM were CD11b+, F4/80+ and CD11c−. IFN-γ (PeproTech), muramyl dipeptide (MDP) (Bachem, King of Prussia, PA), and lipid A (Peptides International, Louisville, KY) were used in vitro.
Wu X., Lahiri A., Haines GK I.I.I., Flavell R.A, & Abraham C. (2014). NOD2 regulates CXCR3-dependent CD8+ T cell accumulation in intestinal tissues with acute injury. Journal of immunology (Baltimore, Md. : 1950), 192(7), 3409-3418.
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3

Human MDMs Cytokine Response to Stimuli

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Human MDMs were treated with lipid A (Peptides International, Louisville, KY), Pam3Cys (EMD Millipore, Billerica, MA), polyinosinic-polycytidylic acid [poly(I:C)], CpG DNA (Invivogen, San Diego, CA), IL-12, IFN-γ (R&D Systems, Minneapolis, MN), IFN-α (Biovision, Milpitas, CA), or cocultured with Salmonella enterica serovar Typhimurium at multiplicity of infection (MOI) 10:1. Supernatants were assayed for IL-6, IL-10, TNF, IL-1β, IFN-γ (BD Biosciences), IL-12 (eBioscience, San Diego, CA), or IFN-γ (BioLegend) by ELISA. In some cases, cells were pretreated with 5 μg/ml fludarabine (STAT1 inhibitor), 150 μM lisofylline (STAT4 inhibitor) (Cayman Chemical Company), Upadacitinib (JAK1 inhibitor), or BMS-986165 (TYK2 inhibitor) (MedChemExpress).
Hedl M., Sun R, & Abraham C. (2020). Disease Risk-Associated Genetic Variants in STAT1 and STAT4 Function in a Complementary Manner to Increase Pattern-Recognition Receptor–Induced Outcomes in Human Macrophages. Journal of immunology (Baltimore, Md. : 1950), 205(5), 1406-1418.
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4

Modulation of Human Macrophage Responses

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Human MDMs were treated with MDP (Bachem, Torrance, CA), lipid A (Peptides International, Louisville, KY), Pam3Cys (EMD Millipore, Billerica, MA), poly(I:C), flagellin, CL097, CpG DNA (Invivogen, San Diego, CA), 2ng/ml TGFβ, 10ng/ml IL4 (R&D Systems, Minneapolis, MN), 10ng/ml IL13 (Biolegend, San Diego, CA), 10ng/ml IL22, 10ng/ml IL10, or 10ng/ml TSLP (Peprotech, Rocky Hill, NJ) unless otherwise stated. In other cases cells were co-cultured with Salmonella enterica serovar Typhimurium at MOI 10:1. For inhibitor studies, cells were pre-incubated with 25μM STAT3 inhibitor peptide or 25μM STAT5 inhibitor (EMD Millipore) (unless otherwise indicated) for 1h before treatments. Supernatants were assayed for cytokine secretion per manufacturer instructions using antibodies to the following proteins: TNF, IL6, IL8, IL10 (BD Biosciences, San Jose, CA), IL12p40 or IL1β (eBioscience, San Diego, CA), TGFβ, IL4, IL13 (Biolegend), IL22 (R&D Systems), or TSLP (PeproTech, Rocky Hill, NJ).
Hedl M., Sun R., Huang C, & Abraham C. (2019). STAT3 and STAT5 signaling thresholds determine distinct regulation for innate-receptor-induced inflammatory cytokines and STAT3/STAT5 disease variants modulate these outcomes. Journal of immunology (Baltimore, Md. : 1950), 203(12), 3325-3338.
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5

Cytokine Secretion in Bone Marrow Cells

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Bone marrow cells were cultured in Dulbecco’s modified Eagle medium containing 10% L929-conditioned medium and used at 6–8 days. Cells were treated with 10 ng/mL recombinant TNFSF15 (Biolegend) or lipid A (Peptides International). In some cases, cells were treated first with neutralizing anti-TNFSF15 antibodies (2 μg/mL, 5G4.6; Bio X Cell, Lebanon, NH) for 30 minutes. Supernatants were assayed for cytokines as follows: TNF (6B8 and MP6-XT22), IL6 (MP5-20F3 and MP5-32C11), IL12/p40 (C15.6 and C17.8), IL10 (JES5-2A5 and JES5-16E3), IL1β (B122) (Biolegend), and biotin-IL1β (eBioscience).
Sun R., Hedl M, & Abraham C. (2020). TNFSF15 Promotes Antimicrobial Pathways in Human Macrophages and These Are Modulated by TNFSF15 Disease-Risk Variants. Cellular and Molecular Gastroenterology and Hepatology, 11(1), 249-272.
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6

Myeloid Cell Isolation and Stimulation

Cited 1 time
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Monocytes were purified from human peripheral blood mononuclear cells by positive CD14 selection (Miltenyi Biotec, Auburn, CA) or by adhesion, tested for purity and cultured with 10 ng ml−1 macrophage colony-stimulating factor (M-CSF; Shenandoah Biotechnology, Warwick, PA; for MDM differentiation) or 40 ng ml−1 GM-CSF and 40 ng ml−1 IL-4 (R&D Systems Inc., Minneapolis, MN; for MDDC differentiation) as in ref. 42 (link). Cultured myeloid cells were treated with MDP (Bachem, King of Prussia, PA), Pam3Cys (EMD Millipore, Billerica, MA), lipid A (Peptides International, Louisville, KY), TriDAP or flagellin (Invivogen, San Diego, CA). In some experiments, MDMs were treated with polyphenol oxidase inhibitors (5 μM kojic acid, 1 mM salicylhydroxamic acid (VWR, Radnor, PA)) or a Syk inhibitor (1 μM; ThermoFisher Scientific, Waltham, MA). Supernatants were assayed for TNF (clones MAb1 and MAb11, diluted 1:1,000), IL-6 (clones MQ2-13A5 and MQ2-39C3, diluted 1:1,000), IL-8 (clones G265-5 and G265-8, diluted 1:1,000), IL-10 (clones JES3-9D7 and JES3-12G8, diluted 1:1,000; BD Biosciences) or IL-1β (clones CRM56 and CRM57 diluted 1:1,000; eBioscience, San Diego, CA) by ELISA. Myeloid cells (CD11c purity >75%) were isolated as in ref. 51 (link) from colonic resection specimens from uninvolved intestine in seven non-IBD patients undergoing surgery for diverticular disease or colon cancer.
Lahiri A., Hedl M., Yan J, & Abraham C. (2017). Human LACC1 increases innate receptor-induced responses and a LACC1 disease-risk variant modulates these outcomes. Nature Communications, 8, 15614.
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7

Myeloid Cell Responses to Microbial Stimuli

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Myeloid-derived cells were treated with muramyl dipeptide (MDP) (Bachem, King of Prussia, Pennsylvania, USA), Pam3Cys-Ser-(Lys)4 (Calbiochem, La Jolla, California, USA), lipid A (Peptides International, Louisville, Kentucky, USA), flagellin, CL097, CpG, poly I:C, Tri-DAP (Invivogen, San Diego, California, USA) or Salmonella enterica serovar Typhimurium (S. typhimurium) at multiplicity of infection (MOI) 10:1. For antibody and inhibitor treatments, cells were incubated with IL-1Ra (IL-1R antagonist) (Genscript), anti-IL-18 receptor accessory protein (RAP) (R&D Systems), Ac-YVAD-Cho (YVAD), caspase-8 inhibitor II or TPL2 inhibitor (Calbiochem) 1 h prior to treatment. Supernatants were assayed for tumour necrosis factor (TNF)-α, IL-8, IL-10 (BD Biosciences), IL-23, IL-1β and IL-18 (eBioscience) by ELISA.
Hedl M, & Abraham C. (2015). A TPL2 (MAP3K8) disease-risk polymorphism increases TPL2 expression thereby leading to increased pattern recognition receptor-initiated caspase-1 and caspase-8 activation, signalling and cytokine secretion. Gut, 65(11), 1799-1811.
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8

Bone Marrow-Derived Dendritic Cell Culture and Cytokine Analysis

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Bone marrow cell suspensions were cultured in complete DMEM containing 20
ng/ml GM-CSF (PeproTech) to generate bone marrow–derived dendritic cells
(BMDCs). Cells were used at 6–8 d. In some cases, BMDCs were first
treated with 0.1 μg/ml lipid A (Peptides International) for 24 h. In
other cases, cells were cocultured with 10 μg/ml OVA-DQ (Invitrogen). In
yet other cases, cells were cultured with 20 μM BAY 11–7082
(NF-κB inhibitor), 10 μM PD98059 (ERK inhibitor), 10 μM
SB202190 (p38 inhibitor) (Calbiochem Research Biochemicals), or 10 μM GSK
2334470 (PDK1 inhibitor) (Tocris). The following were assessed by ELISA: IL-6,
IL-12p40, and IL-23 (eBioscience); TNF (BioLegend); IL-1β, CCL2, CXCL2,
CXCL9, and CXCL10 (PeproTech); and CCL19, CCL21, and CXCL9 (R&D
Systems).
Yan J., Hedl M, & Abraham C. (2020). Myeloid Cell–Intrinsic IRF5 Promotes T Cell Responses through Multiple Distinct Checkpoints In Vivo, and IRF5 Immune-Mediated Disease Risk Variants Modulate These Myeloid Cell Functions. Journal of immunology (Baltimore, Md. : 1950), 205(4), 1024-1038.
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9

Bone Marrow Macrophage Stimulation

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Bone marrow cell suspensions were cultured in complete DMEM containing 10% L929-conditioned medium. Cells were used at 6 to 8 days. In some cases BMMs were first treated with lipid A (Peptides International) for 48 h. In other cases BMMs were cultured with S. Typhimurium, Adherent Invasive Escherichia coli (AIEC) strain LF82 (generously from Dr. Emiko Mizoguchi), or Staphylococcus aureus at 5:1 MOI for 24 h, with 30 μg/ml gentamycin added after the first 20 min.
Pandey S.P., Yan J., Turner J.R, & Abraham C. (2019). Reducing IRF5 expression attenuates colitis in mice, but impairs the clearance of intestinal pathogens. Mucosal immunology, 12(4), 874-887.
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10

Cytokine Secretion in Human MDMs

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Human MDMs were treated with MDP (Bachem, Torrance, CA), Pam3Cys (MilliporeSigma, Burlington, MA), lipid A (Peptides International, Louisville, KY), CpG DNA (InvivoGen, San Diego, CA), or TNFSF15 (Peprotech, Rocky Hill, NJ). In some cases, cells were given neutralizing anti-DR3 antibodies (1 μg/mL; JD3; ThermoFisher Scientific, Waltham, MA) for 1 hour before treatments. In other cases, cells were given 10 μmol/L TAPI-1 (MilliporeSigma) for 1 hour before treatments. Supernatants were assayed for cytokine secretion per the manufacturer's instructions using antibodies to the following proteins: TNFSF15 (Peprotech), TNF (Mab1 and Mab11), IL6 (MQ2-13A5 and MQ2-39C3), IL8 (G265-5 and G265-8), IL10 (JES3-9D7 and JES3-12G8) (BD Biosciences, San Jose, CA), or IL1β (CRM56 and CRM57) (eBioscience, San Diego, CA).
Sun R., Hedl M, & Abraham C. (2020). TNFSF15 Promotes Antimicrobial Pathways in Human Macrophages and These Are Modulated by TNFSF15 Disease-Risk Variants. Cellular and Molecular Gastroenterology and Hepatology, 11(1), 249-272.
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