Journal: The Journal of Immunology Author Choice

Article Title: The Regulation of Nucleic Acid Vaccine Responses by the Microbiome

doi: 10.4049/jimmunol.2300196

Figure Lengend Snippet: Details of Ab reagents used in these experiments

Article Snippet: Ab target Conjugation Clone Manufacturer Product Number Concentration OVA-specific IgG1 None L71 Chondrex 7093 OVA-specific IgG2b None 4B436 Chondrex 7096 OVA-specific IgG2c None 3E3A9 Chondrex 7109 Mouse IgG HRP Poly4053 BioLegend 405306 1:3000 dilution Mouse IgG1 HRP — Invitrogen A10551 0.5 μg/ml Mouse IgG2b HRP — Invitrogen M32507 0.5 μg/ml Mouse IgG2c HRP — Invitrogen PAI-29288 0.5 μg/ml Mouse CD16/32 None 93 SouthernBiotech, BioLegend 1630-01 1 μg/ml Mouse CD3 BUV395 145-2C11 BD Biosciences 563565 2 μg/ml Mouse CD4 PerCP-eFluor710 RM4-5 Invitrogen 46-0042-82 1 μg/ml Mouse CD8 BUV737 53-6.7 BD Biosciences 612739 1 μg/ml Mouse IFN-γ Alexa Fluor 488 XMG1.2 BioLegend 505813 2 μg/ml Mouse TNF-α Pacific Blue MP6-XT22 BioLegend 506318 2 μg/ml Mouse IL-17 PE TC11-18H10 BioLegend 506904 1 μg/ml Mouse IL-4 PE-CF594 11B11 BioLegend 504132 1 μg/ml Mouse IL-10 APC JES5-16E3 BioLegend 505010 1 μg/ml Mouse CD11b BUV395 M1/70 BD Biosciences 563553 1 μg/ml Mouse CD11c FITC N418 BioLegend 117306 5 μg/ml Mouse Ly6G BV510 1A8 BioLegend 127633 1 μg/ml Mouse Ly6C BV785 HK1.4 BioLegend 128041 1 μg/ml Mouse CD103 PECy7 2E7 BioLegend 121426 1 μg/ml Mouse CD8a BUV730 53-6.7 BD Biosciences 612759 1 μg/ml Mouse XCR1 APCCy7 ZET BioLegend 148223 1 μg/ml Mouse CD301b EF660 11A10-B7 Invitrogen 50-3011-80 1 μg/ml Mouse CD64 BV605 X54-5/7.1 BioLegend 139323 1 μg/ml Mouse CD80 BV650 16-10AE BioLegend 104732 1 μg/ml Mouse CD86 PE PO3.1 eBioscience 12-0861-82 1 μg/ml Mouse MHC-II (I-A/I-E) AF700 M5/114.15.2 Invitrogen 565321-82 1 μg/ml CD45 BUV805 30-F112 BD Biosciences 748370 1 μg/ml CD107a FITC 1D4B BD Biosciences 553793 5 μg/ml CD4 PerCP-Cy5.5 GK1.5 BioLegend 100434 2 μg/ml CD8a Pacific Blue 53-6.7 BioLegend 100725 5 μg/ml CD3e BV605 145-2C11 BioLegend 100351 2 μg/ml IL-4 BV786 11B11 BD Biosciences 564006 2 μg/ml IL-2 APC JES6-5H4 BD Biosciences 554429 2 μg/ml IFN-γ AF700 XMG1.2 BD Biosciences 557998 2 μg/ml IL-17 APC-Cy7 TC11-18H10.1 BioLegend 506940 2 μg/ml IL-5 PE TRFK5 BioLegend 504304 2 μg/ml TNF-α PE-Cy7 MP6-XT22 BD Biosciences 557644 2 μg/ml Open in a separate window Details of Ab reagents used in these experiments

Techniques: Conjugation Assay, Concentration Assay

Journal: The Journal of Immunology Author Choice

Article Title: The Regulation of Nucleic Acid Vaccine Responses by the Microbiome

doi: 10.4049/jimmunol.2300196

Figure Lengend Snippet: Flow cytometry profiling of innate immune response to mRNA-LNP in GF and SPF mice. (A) Representative flow cytometry and graphical summaries of Ly6Chi monocyte gating and monocyte CD11c versus MHC class II expression in PBS-treated or mRNA-LNP-immunized GF and SPF mice. (B) Gating strategy to identify neutrophils and Ly6C+, CD64+, and CD64−Ly6C− DC/macrophage populations among splenocytes from PBS-treated or mRNA-LNP-immunized GF and SPF mice. (C) Graphical summaries of neutrophil and Ly6C+, CD64+, and CD64− Ly6C− DC/macrophage proportions in PBS-treated or mRNA-LNP–immunized mice. (D) Median fluorescent intensity (MFI) for CD86 and CD80 expression on the specified populations of immune cells from the spleens of GF or SPF mice 24 h after immunization with mRNA-LNP vaccine or PBS control. Each data point on graphs represents a single mouse (n = 3 per PBS group, n = 4 per mRNA-LNP group from one experiment). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by one-way ANOVA.

Article Snippet: Ab target Conjugation Clone Manufacturer Product Number Concentration OVA-specific IgG1 None L71 Chondrex 7093 OVA-specific IgG2b None 4B436 Chondrex 7096 OVA-specific IgG2c None 3E3A9 Chondrex 7109 Mouse IgG HRP Poly4053 BioLegend 405306 1:3000 dilution Mouse IgG1 HRP — Invitrogen A10551 0.5 μg/ml Mouse IgG2b HRP — Invitrogen M32507 0.5 μg/ml Mouse IgG2c HRP — Invitrogen PAI-29288 0.5 μg/ml Mouse CD16/32 None 93 SouthernBiotech, BioLegend 1630-01 1 μg/ml Mouse CD3 BUV395 145-2C11 BD Biosciences 563565 2 μg/ml Mouse CD4 PerCP-eFluor710 RM4-5 Invitrogen 46-0042-82 1 μg/ml Mouse CD8 BUV737 53-6.7 BD Biosciences 612739 1 μg/ml Mouse IFN-γ Alexa Fluor 488 XMG1.2 BioLegend 505813 2 μg/ml Mouse TNF-α Pacific Blue MP6-XT22 BioLegend 506318 2 μg/ml Mouse IL-17 PE TC11-18H10 BioLegend 506904 1 μg/ml Mouse IL-4 PE-CF594 11B11 BioLegend 504132 1 μg/ml Mouse IL-10 APC JES5-16E3 BioLegend 505010 1 μg/ml Mouse CD11b BUV395 M1/70 BD Biosciences 563553 1 μg/ml Mouse CD11c FITC N418 BioLegend 117306 5 μg/ml Mouse Ly6G BV510 1A8 BioLegend 127633 1 μg/ml Mouse Ly6C BV785 HK1.4 BioLegend 128041 1 μg/ml Mouse CD103 PECy7 2E7 BioLegend 121426 1 μg/ml Mouse CD8a BUV730 53-6.7 BD Biosciences 612759 1 μg/ml Mouse XCR1 APCCy7 ZET BioLegend 148223 1 μg/ml Mouse CD301b EF660 11A10-B7 Invitrogen 50-3011-80 1 μg/ml Mouse CD64 BV605 X54-5/7.1 BioLegend 139323 1 μg/ml Mouse CD80 BV650 16-10AE BioLegend 104732 1 μg/ml Mouse CD86 PE PO3.1 eBioscience 12-0861-82 1 μg/ml Mouse MHC-II (I-A/I-E) AF700 M5/114.15.2 Invitrogen 565321-82 1 μg/ml CD45 BUV805 30-F112 BD Biosciences 748370 1 μg/ml CD107a FITC 1D4B BD Biosciences 553793 5 μg/ml CD4 PerCP-Cy5.5 GK1.5 BioLegend 100434 2 μg/ml CD8a Pacific Blue 53-6.7 BioLegend 100725 5 μg/ml CD3e BV605 145-2C11 BioLegend 100351 2 μg/ml IL-4 BV786 11B11 BD Biosciences 564006 2 μg/ml IL-2 APC JES6-5H4 BD Biosciences 554429 2 μg/ml IFN-γ AF700 XMG1.2 BD Biosciences 557998 2 μg/ml IL-17 APC-Cy7 TC11-18H10.1 BioLegend 506940 2 μg/ml IL-5 PE TRFK5 BioLegend 504304 2 μg/ml TNF-α PE-Cy7 MP6-XT22 BD Biosciences 557644 2 μg/ml Open in a separate window Details of Ab reagents used in these experiments

Techniques: Flow Cytometry, Expressing, Control