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mip 1α  (R&D Systems)


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    R&D Systems mip 1α
    Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. ( A ) Schematic of training assay. ( B – G ) Expression of HLA-DR ( B ), CD14 ( C ), CD40 ( D ), CD80 ( E ), H3K27ac ( F ), or H3K4me3 ( G ) on unstimulated MDM on day 7 measured by flow cytometry. MFI, median fluorescence intensity. ( H and I ) Expression (relative to untrained) of NFKB1 ( H ) or IRF1 ( I ) in unstimulated MDM on day 7 measured by qPCR. ( J – O ) On day 6, MDM were stimulated with LPS (10 ng/mL) or irradiated M.tb (10 μg/mL) for 24 hours, and TNF ( J ), IL-6 ( K ), IL-1β ( L ), IL-10 ( M ), CXCL1 ( N ), <t>or</t> <t>MIP-1α</t> ( O ) was measured by ELISA. Each dot represents an individual donor, n = 6 ( B – E , H , and I ), n = 5 ( F and G ), or n = 7 ( J – O ), with paired data joined by a line. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined using a paired t test ( B – I ) or a 2-way ANOVA with Šidák’s multiple-comparison test ( J – O ).
    Mip 1α, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+mip+1%CE%B1/Human+CCL3%2FMIP-1+alpha+DuoSet+ELISA/pmc13043095-282-17-18
    Average 94 stars, based on 32 article reviews
    mip 1α - by Bioz Stars, 2026-09
    94/100 stars

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    1) Product Images from "IFN- γ –induced trained immunity enhances killing of priority pathogens in healthy and genetically vulnerable individuals"

    Article Title: IFN- γ –induced trained immunity enhances killing of priority pathogens in healthy and genetically vulnerable individuals

    Journal: JCI Insight

    doi: 10.1172/jci.insight.195866

    Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. ( A ) Schematic of training assay. ( B – G ) Expression of HLA-DR ( B ), CD14 ( C ), CD40 ( D ), CD80 ( E ), H3K27ac ( F ), or H3K4me3 ( G ) on unstimulated MDM on day 7 measured by flow cytometry. MFI, median fluorescence intensity. ( H and I ) Expression (relative to untrained) of NFKB1 ( H ) or IRF1 ( I ) in unstimulated MDM on day 7 measured by qPCR. ( J – O ) On day 6, MDM were stimulated with LPS (10 ng/mL) or irradiated M.tb (10 μg/mL) for 24 hours, and TNF ( J ), IL-6 ( K ), IL-1β ( L ), IL-10 ( M ), CXCL1 ( N ), or MIP-1α ( O ) was measured by ELISA. Each dot represents an individual donor, n = 6 ( B – E , H , and I ), n = 5 ( F and G ), or n = 7 ( J – O ), with paired data joined by a line. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined using a paired t test ( B – I ) or a 2-way ANOVA with Šidák’s multiple-comparison test ( J – O ).
    Figure Legend Snippet: Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. ( A ) Schematic of training assay. ( B – G ) Expression of HLA-DR ( B ), CD14 ( C ), CD40 ( D ), CD80 ( E ), H3K27ac ( F ), or H3K4me3 ( G ) on unstimulated MDM on day 7 measured by flow cytometry. MFI, median fluorescence intensity. ( H and I ) Expression (relative to untrained) of NFKB1 ( H ) or IRF1 ( I ) in unstimulated MDM on day 7 measured by qPCR. ( J – O ) On day 6, MDM were stimulated with LPS (10 ng/mL) or irradiated M.tb (10 μg/mL) for 24 hours, and TNF ( J ), IL-6 ( K ), IL-1β ( L ), IL-10 ( M ), CXCL1 ( N ), or MIP-1α ( O ) was measured by ELISA. Each dot represents an individual donor, n = 6 ( B – E , H , and I ), n = 5 ( F and G ), or n = 7 ( J – O ), with paired data joined by a line. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined using a paired t test ( B – I ) or a 2-way ANOVA with Šidák’s multiple-comparison test ( J – O ).

    Techniques Used: Expressing, Flow Cytometry, Fluorescence, Irradiation, Enzyme-linked Immunosorbent Assay, Comparison

    Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. On day 7, MDM were infected with M . tuberculosis (H37Rv; MOI 1–5) ( A and B ), stimulated with irradiated M.tb (10 μg/mL) ( G ), or infected with S . aureus (USA300; MOI 100:1) ( H – P ) for the indicated times. ( A and B ) Bacterial M . tuberculosis burden within MDM 3 and 48 hours after infection. ( C – F ) Relative expression (to untrained) of CYBA ( C ), CYBB ( D ), NCF1 ( E ), or NCF2 ( F ) in uninfected MDM on day 7 (qPCR). ( G ) MDM were stimulated with irradiated M.tb for 4 hours, and ROS (DHR123; relative to unstimulated MDM) was measured (flow cytometry). ( H – K ) TNF ( H ), IL-6 ( I ), IL-1β ( J ), or IL-10 ( K ) (ELISA) 24 hours after S . aureus infection. ( L ) MDM infected with CFSE-labeled S . aureus (%). ( M ) Intracellular bacterial S . aureus burden within MDM 1, 4, or 24 hours after gentamicin. ( N ) Spearman’s r correlation matrix: correlation of reduced CFU with IL-1β, TNF, IL-6, IL-10, CXCL1, or MIP-1α production. ( O ) MDM were infected with S . aureus alone or in the presence of NAC (10 mM) for 4 hours, and ROS was measured (flow cytometry). ( P ) Fold change in CFU/mL of untrained or IFN-γ–trained (stripes) MDM infected with S . aureus 1, 4, or 24 hours after gentamicin in healthy controls (HC; white) versus MDM from a patient with chronic granulomatous disease (CGD; green). Each dot represents an individual donor, n = 4 ( A – G ), n = 7 ( H – K , M , and N ), n = 5 ( L ), n = 6 ( O ), or HC n = 7, CGD n = 1 ( P ). Data are graphed as paired data joined by a line or the mean value ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, paired t test ( A – L ) or 2-way ANOVA with Šidák’s multiple-comparison test ( M ), uncorrected Fisher’s LSD test ( O ), or Tukey’s multiple-comparison test ( P ).
    Figure Legend Snippet: Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. On day 7, MDM were infected with M . tuberculosis (H37Rv; MOI 1–5) ( A and B ), stimulated with irradiated M.tb (10 μg/mL) ( G ), or infected with S . aureus (USA300; MOI 100:1) ( H – P ) for the indicated times. ( A and B ) Bacterial M . tuberculosis burden within MDM 3 and 48 hours after infection. ( C – F ) Relative expression (to untrained) of CYBA ( C ), CYBB ( D ), NCF1 ( E ), or NCF2 ( F ) in uninfected MDM on day 7 (qPCR). ( G ) MDM were stimulated with irradiated M.tb for 4 hours, and ROS (DHR123; relative to unstimulated MDM) was measured (flow cytometry). ( H – K ) TNF ( H ), IL-6 ( I ), IL-1β ( J ), or IL-10 ( K ) (ELISA) 24 hours after S . aureus infection. ( L ) MDM infected with CFSE-labeled S . aureus (%). ( M ) Intracellular bacterial S . aureus burden within MDM 1, 4, or 24 hours after gentamicin. ( N ) Spearman’s r correlation matrix: correlation of reduced CFU with IL-1β, TNF, IL-6, IL-10, CXCL1, or MIP-1α production. ( O ) MDM were infected with S . aureus alone or in the presence of NAC (10 mM) for 4 hours, and ROS was measured (flow cytometry). ( P ) Fold change in CFU/mL of untrained or IFN-γ–trained (stripes) MDM infected with S . aureus 1, 4, or 24 hours after gentamicin in healthy controls (HC; white) versus MDM from a patient with chronic granulomatous disease (CGD; green). Each dot represents an individual donor, n = 4 ( A – G ), n = 7 ( H – K , M , and N ), n = 5 ( L ), n = 6 ( O ), or HC n = 7, CGD n = 1 ( P ). Data are graphed as paired data joined by a line or the mean value ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, paired t test ( A – L ) or 2-way ANOVA with Šidák’s multiple-comparison test ( M ), uncorrected Fisher’s LSD test ( O ), or Tukey’s multiple-comparison test ( P ).

    Techniques Used: Infection, Irradiation, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Labeling, Comparison

    Related Articles

    Recombinant:

    Article Title: CCR1 expression and signal transduction by murine BMMC results in secretion of TNF-α, TGFβ-1 and IL-6
    Article Snippet: Immunoreactivity was visualized by applying HRP ECL substrate (Pierce, Rockford, IL, USA) and immediately exposing the membranes to x-ray film. .. Resting and sensitized BMMC were stimulated for 30 min with 10.0 ng ml −1 dinitrylphenyl human serum albumin (DNP-HSA; Sigma) and/or various concentrations of recombinant human MIP-1α (R&D Systems) in Tyrode's buffer (DMEM containing 0.1% BSA and 10 mM HEPES). .. Total cellular RNA was extracted from 5 × 10 6 BMMC using the RNeasy kit (Qiagen, Valencia, CA, USA), following the manufacturer's instructions.

    Article Title: GWAS-identified CCR1 and IL10 loci contribute to M1 macrophage-predominant inflammation in Behçet’s disease
    Article Snippet: .. In the lower chamber, 1% bovine serum albumin (Wako Pure Chemical Co., Osaka, Japan)-supplemented RPMI 1640 medium with or without recombinant human MIP-1α (R&D Systems) was applied. .. After incubation for 2 hours, cells in the upper chamber were removed with Q-Tips, and cells on the bottom side of the membrane were stained with Diff-Quik (Sysmex, Kobe, Japan).

    Article Title: Molecules and methods of using same for treating CCR5/CCR5 ligands associated diseases
    Article Snippet: Chemotaxis assays were conducted using a TransWell chamber (Corning Costar, Cambridge, Mass.). .. THP-1 cells with medium (1×106 cells/well) were added to the upper chamber of the Transwell, after equilibration of the lower chambers with medium or with recombinant human MIP-1α (CCL3, R&D Systems, Minneapolis, N. ..

    Article Title: Adsorbent and method for adsorbing a chemokine in body fluid
    Article Snippet: .. Three ml of MIP-1α added-normal human serum (the concentration of MIP-1α: 1.1 ng/ml) which was prepared by adding recombinant human MIP-1α (made by R & D Systems Corporation) to a normal human serum (made by Dainippon Pharmaceutical Co., Ltd.) was added to each of 0.5 ml of this immobilized carrier and 0.5 ml of CELLULOFINE GC-700 m, and each mixture was incubated with shaking at 37° C. for two hours. .. Concentrations of MIP-1α in the supernatant before and after the incubation were measured by using a kit for measuring human MIP-1α made by R & D SYSTEMS Corporation, and the adsorption rate was calculated by the following formula: Adsorption rate ⁢ ⁢ ( % ) = ( concentration in ⁢ ⁢ serum ⁢ ⁢ before incubation ) - ( concentration in ⁢ ⁢ serum ⁢ ⁢ after incubation ) ( concentration ⁢ ⁢ in ⁢ ⁢ serum before ⁢ ⁢ incubation ) × 100 It is noted that the concentration in serum before incubation is a value corrected for the water contained in the carrier.

    Article Title: GWAS-identified CCR1 and IL10 loci contribute to M1 macrophage-predominant inflammation in Behçet's disease.
    Article Snippet: .. In the lower chamber, 1% bovine serum albumin (Wako Pure Chemical Co., Osaka, Japan)-supplemented RPMI 1640 medium with or without recombinant human MIP-1α (R&D Systems) was applied. .. After incubation for 2 hours, cells in the upper chamber were removed with Q-Tips, and cells on the bottom side of the membrane were stained with Diff-Quik (Sysmex, Kobe, Japan).

    Labeling:

    Article Title: Discovery and Lead Optimization of a Novel Series of CC Chemokine Receptor 1 (CCR1)-Selective Piperidine Antagonists via Parallel Synthesis
    Article Snippet: A series of novel, potent CCR1 inhibitors was developed from a moderately active hit using an iterative parallel synthesis approach.. The initial hit (composed of three subunits: an amine, a central amino acid, and an N-terminal cap) became the basis for a series of parallel chemical libraries designed to generate SAR data.. Libraries were synthesized that explored each of the three subunits; the CCR1 binding data obtained revealed the following: (1) changes to the amine are not well tolerated; (2) small alkylamino acids are preferred in the center of the molecule; (3) substitutions at the N-terminus are generally well tolerated.

    Chemotaxis Assay:

    Article Title: Discovery and Lead Optimization of a Novel Series of CC Chemokine Receptor 1 (CCR1)-Selective Piperidine Antagonists via Parallel Synthesis
    Article Snippet: A series of novel, potent CCR1 inhibitors was developed from a moderately active hit using an iterative parallel synthesis approach.. The initial hit (composed of three subunits: an amine, a central amino acid, and an N-terminal cap) became the basis for a series of parallel chemical libraries designed to generate SAR data.. Libraries were synthesized that explored each of the three subunits; the CCR1 binding data obtained revealed the following: (1) changes to the amine are not well tolerated; (2) small alkylamino acids are preferred in the center of the molecule; (3) substitutions at the N-terminus are generally well tolerated.

    Pore Size:

    Article Title: Discovery and Lead Optimization of a Novel Series of CC Chemokine Receptor 1 (CCR1)-Selective Piperidine Antagonists via Parallel Synthesis
    Article Snippet: A series of novel, potent CCR1 inhibitors was developed from a moderately active hit using an iterative parallel synthesis approach.. The initial hit (composed of three subunits: an amine, a central amino acid, and an N-terminal cap) became the basis for a series of parallel chemical libraries designed to generate SAR data.. Libraries were synthesized that explored each of the three subunits; the CCR1 binding data obtained revealed the following: (1) changes to the amine are not well tolerated; (2) small alkylamino acids are preferred in the center of the molecule; (3) substitutions at the N-terminus are generally well tolerated.

    BrdU Cell Proliferation Assay:

    Article Title: Secreted tryptophanyl-tRNA synthetase as a primary defence system against infection.
    Article Snippet: Young Ha Ahn1†, Sunyoung Park2†, Jeong June Choi2, Bo-Kyung Park2, Kyung Hee Rhee3, Eunjoo Kang4, Soyeon Ahn5, Chul-Ho Lee6, Jong Soo Lee7, Kyung-Soo Inn8, Mi-La Cho9, Sung-Hwan Park10, Kyunghee Park11,12, Hae Jung Park11,12, Jae Hyun Lee11,12, Jung Won Park11,12, Nam Hoon Kwon4, Hyunbo Shim13, Byung Woo Han3, Pilhan Kim5, Joo-Youn Lee1,14, Youngho Jeon15, Jin Won Huh16, Mirim Jin* and Sunghoon Kim*

    Phagocytosis Assay:

    Article Title: Secreted tryptophanyl-tRNA synthetase as a primary defence system against infection.
    Article Snippet: Young Ha Ahn1†, Sunyoung Park2†, Jeong June Choi2, Bo-Kyung Park2, Kyung Hee Rhee3, Eunjoo Kang4, Soyeon Ahn5, Chul-Ho Lee6, Jong Soo Lee7, Kyung-Soo Inn8, Mi-La Cho9, Sung-Hwan Park10, Kyunghee Park11,12, Hae Jung Park11,12, Jae Hyun Lee11,12, Jung Won Park11,12, Nam Hoon Kwon4, Hyunbo Shim13, Byung Woo Han3, Pilhan Kim5, Joo-Youn Lee1,14, Youngho Jeon15, Jin Won Huh16, Mirim Jin* and Sunghoon Kim*

    Enzyme-linked Immunosorbent Assay:

    Article Title: Secreted tryptophanyl-tRNA synthetase as a primary defence system against infection.
    Article Snippet: Young Ha Ahn1†, Sunyoung Park2†, Jeong June Choi2, Bo-Kyung Park2, Kyung Hee Rhee3, Eunjoo Kang4, Soyeon Ahn5, Chul-Ho Lee6, Jong Soo Lee7, Kyung-Soo Inn8, Mi-La Cho9, Sung-Hwan Park10, Kyunghee Park11,12, Hae Jung Park11,12, Jae Hyun Lee11,12, Jung Won Park11,12, Nam Hoon Kwon4, Hyunbo Shim13, Byung Woo Han3, Pilhan Kim5, Joo-Youn Lee1,14, Youngho Jeon15, Jin Won Huh16, Mirim Jin* and Sunghoon Kim*

    Concentration Assay:

    Article Title: Adsorbent and method for adsorbing a chemokine in body fluid
    Article Snippet: .. Three ml of MIP-1α added-normal human serum (the concentration of MIP-1α: 1.1 ng/ml) which was prepared by adding recombinant human MIP-1α (made by R & D Systems Corporation) to a normal human serum (made by Dainippon Pharmaceutical Co., Ltd.) was added to each of 0.5 ml of this immobilized carrier and 0.5 ml of CELLULOFINE GC-700 m, and each mixture was incubated with shaking at 37° C. for two hours. .. Concentrations of MIP-1α in the supernatant before and after the incubation were measured by using a kit for measuring human MIP-1α made by R & D SYSTEMS Corporation, and the adsorption rate was calculated by the following formula: Adsorption rate ⁢ ⁢ ( % ) = ( concentration in ⁢ ⁢ serum ⁢ ⁢ before incubation ) - ( concentration in ⁢ ⁢ serum ⁢ ⁢ after incubation ) ( concentration ⁢ ⁢ in ⁢ ⁢ serum before ⁢ ⁢ incubation ) × 100 It is noted that the concentration in serum before incubation is a value corrected for the water contained in the carrier.

    Incubation:

    Article Title: Adsorbent and method for adsorbing a chemokine in body fluid
    Article Snippet: .. Three ml of MIP-1α added-normal human serum (the concentration of MIP-1α: 1.1 ng/ml) which was prepared by adding recombinant human MIP-1α (made by R & D Systems Corporation) to a normal human serum (made by Dainippon Pharmaceutical Co., Ltd.) was added to each of 0.5 ml of this immobilized carrier and 0.5 ml of CELLULOFINE GC-700 m, and each mixture was incubated with shaking at 37° C. for two hours. .. Concentrations of MIP-1α in the supernatant before and after the incubation were measured by using a kit for measuring human MIP-1α made by R & D SYSTEMS Corporation, and the adsorption rate was calculated by the following formula: Adsorption rate ⁢ ⁢ ( % ) = ( concentration in ⁢ ⁢ serum ⁢ ⁢ before incubation ) - ( concentration in ⁢ ⁢ serum ⁢ ⁢ after incubation ) ( concentration ⁢ ⁢ in ⁢ ⁢ serum before ⁢ ⁢ incubation ) × 100 It is noted that the concentration in serum before incubation is a value corrected for the water contained in the carrier.



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    Image Search Results


    Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. ( A ) Schematic of training assay. ( B – G ) Expression of HLA-DR ( B ), CD14 ( C ), CD40 ( D ), CD80 ( E ), H3K27ac ( F ), or H3K4me3 ( G ) on unstimulated MDM on day 7 measured by flow cytometry. MFI, median fluorescence intensity. ( H and I ) Expression (relative to untrained) of NFKB1 ( H ) or IRF1 ( I ) in unstimulated MDM on day 7 measured by qPCR. ( J – O ) On day 6, MDM were stimulated with LPS (10 ng/mL) or irradiated M.tb (10 μg/mL) for 24 hours, and TNF ( J ), IL-6 ( K ), IL-1β ( L ), IL-10 ( M ), CXCL1 ( N ), or MIP-1α ( O ) was measured by ELISA. Each dot represents an individual donor, n = 6 ( B – E , H , and I ), n = 5 ( F and G ), or n = 7 ( J – O ), with paired data joined by a line. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined using a paired t test ( B – I ) or a 2-way ANOVA with Šidák’s multiple-comparison test ( J – O ).

    Journal: JCI Insight

    Article Title: IFN- γ –induced trained immunity enhances killing of priority pathogens in healthy and genetically vulnerable individuals

    doi: 10.1172/jci.insight.195866

    Figure Lengend Snippet: Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. ( A ) Schematic of training assay. ( B – G ) Expression of HLA-DR ( B ), CD14 ( C ), CD40 ( D ), CD80 ( E ), H3K27ac ( F ), or H3K4me3 ( G ) on unstimulated MDM on day 7 measured by flow cytometry. MFI, median fluorescence intensity. ( H and I ) Expression (relative to untrained) of NFKB1 ( H ) or IRF1 ( I ) in unstimulated MDM on day 7 measured by qPCR. ( J – O ) On day 6, MDM were stimulated with LPS (10 ng/mL) or irradiated M.tb (10 μg/mL) for 24 hours, and TNF ( J ), IL-6 ( K ), IL-1β ( L ), IL-10 ( M ), CXCL1 ( N ), or MIP-1α ( O ) was measured by ELISA. Each dot represents an individual donor, n = 6 ( B – E , H , and I ), n = 5 ( F and G ), or n = 7 ( J – O ), with paired data joined by a line. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 determined using a paired t test ( B – I ) or a 2-way ANOVA with Šidák’s multiple-comparison test ( J – O ).

    Article Snippet: The concentrations of IL-1β (BioLegend, 437016), IL-10 (BioLegend, 430604), IL-6 (BioLegend, 430516), IFN-γ (BioLegend, 430116), TNF (Invitrogen), MIP-1α (R&D Systems, DY270), and CXCL1 (R&D Systems, DY275) were determined by ELISA following each manufacturer’s instructions.

    Techniques: Expressing, Flow Cytometry, Fluorescence, Irradiation, Enzyme-linked Immunosorbent Assay, Comparison

    Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. On day 7, MDM were infected with M . tuberculosis (H37Rv; MOI 1–5) ( A and B ), stimulated with irradiated M.tb (10 μg/mL) ( G ), or infected with S . aureus (USA300; MOI 100:1) ( H – P ) for the indicated times. ( A and B ) Bacterial M . tuberculosis burden within MDM 3 and 48 hours after infection. ( C – F ) Relative expression (to untrained) of CYBA ( C ), CYBB ( D ), NCF1 ( E ), or NCF2 ( F ) in uninfected MDM on day 7 (qPCR). ( G ) MDM were stimulated with irradiated M.tb for 4 hours, and ROS (DHR123; relative to unstimulated MDM) was measured (flow cytometry). ( H – K ) TNF ( H ), IL-6 ( I ), IL-1β ( J ), or IL-10 ( K ) (ELISA) 24 hours after S . aureus infection. ( L ) MDM infected with CFSE-labeled S . aureus (%). ( M ) Intracellular bacterial S . aureus burden within MDM 1, 4, or 24 hours after gentamicin. ( N ) Spearman’s r correlation matrix: correlation of reduced CFU with IL-1β, TNF, IL-6, IL-10, CXCL1, or MIP-1α production. ( O ) MDM were infected with S . aureus alone or in the presence of NAC (10 mM) for 4 hours, and ROS was measured (flow cytometry). ( P ) Fold change in CFU/mL of untrained or IFN-γ–trained (stripes) MDM infected with S . aureus 1, 4, or 24 hours after gentamicin in healthy controls (HC; white) versus MDM from a patient with chronic granulomatous disease (CGD; green). Each dot represents an individual donor, n = 4 ( A – G ), n = 7 ( H – K , M , and N ), n = 5 ( L ), n = 6 ( O ), or HC n = 7, CGD n = 1 ( P ). Data are graphed as paired data joined by a line or the mean value ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, paired t test ( A – L ) or 2-way ANOVA with Šidák’s multiple-comparison test ( M ), uncorrected Fisher’s LSD test ( O ), or Tukey’s multiple-comparison test ( P ).

    Journal: JCI Insight

    Article Title: IFN- γ –induced trained immunity enhances killing of priority pathogens in healthy and genetically vulnerable individuals

    doi: 10.1172/jci.insight.195866

    Figure Lengend Snippet: Enriched monocytes were left untrained (UT; white) or trained with IFN-γ (10 ng/mL; gray) for 24 hours. Cells were differentiated into MDM. On day 7, MDM were infected with M . tuberculosis (H37Rv; MOI 1–5) ( A and B ), stimulated with irradiated M.tb (10 μg/mL) ( G ), or infected with S . aureus (USA300; MOI 100:1) ( H – P ) for the indicated times. ( A and B ) Bacterial M . tuberculosis burden within MDM 3 and 48 hours after infection. ( C – F ) Relative expression (to untrained) of CYBA ( C ), CYBB ( D ), NCF1 ( E ), or NCF2 ( F ) in uninfected MDM on day 7 (qPCR). ( G ) MDM were stimulated with irradiated M.tb for 4 hours, and ROS (DHR123; relative to unstimulated MDM) was measured (flow cytometry). ( H – K ) TNF ( H ), IL-6 ( I ), IL-1β ( J ), or IL-10 ( K ) (ELISA) 24 hours after S . aureus infection. ( L ) MDM infected with CFSE-labeled S . aureus (%). ( M ) Intracellular bacterial S . aureus burden within MDM 1, 4, or 24 hours after gentamicin. ( N ) Spearman’s r correlation matrix: correlation of reduced CFU with IL-1β, TNF, IL-6, IL-10, CXCL1, or MIP-1α production. ( O ) MDM were infected with S . aureus alone or in the presence of NAC (10 mM) for 4 hours, and ROS was measured (flow cytometry). ( P ) Fold change in CFU/mL of untrained or IFN-γ–trained (stripes) MDM infected with S . aureus 1, 4, or 24 hours after gentamicin in healthy controls (HC; white) versus MDM from a patient with chronic granulomatous disease (CGD; green). Each dot represents an individual donor, n = 4 ( A – G ), n = 7 ( H – K , M , and N ), n = 5 ( L ), n = 6 ( O ), or HC n = 7, CGD n = 1 ( P ). Data are graphed as paired data joined by a line or the mean value ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, paired t test ( A – L ) or 2-way ANOVA with Šidák’s multiple-comparison test ( M ), uncorrected Fisher’s LSD test ( O ), or Tukey’s multiple-comparison test ( P ).

    Article Snippet: The concentrations of IL-1β (BioLegend, 437016), IL-10 (BioLegend, 430604), IL-6 (BioLegend, 430516), IFN-γ (BioLegend, 430116), TNF (Invitrogen), MIP-1α (R&D Systems, DY270), and CXCL1 (R&D Systems, DY275) were determined by ELISA following each manufacturer’s instructions.

    Techniques: Infection, Irradiation, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Labeling, Comparison