Antigen-level resolution of commensal-specific B cell responses can be enabled by phage display screening coupled with B cell tetramers.
Induction of commensal-specific immunity contributes to tissue homeostasis, yet the mechanisms underlying induction of commensal-specific B cells remain poorly understood in part due to a lack of tools to identify these cells. Using phage display, we identified segmented filamentous bacteria (SFB) antigens targeted by serum and intestinal antibodies and generated B cell tetramers to track SFB-specific B cells in gut-associated lymphoid tissues. We revealed a compartmentalized response in SFB-specific B cell activation, with a gradient of immunoglobulin A (IgA), IgG1, and IgG2b isotype production along Peyer's patches contrasted by selective production of IgG2b within mesenteric lymph nodes. V(D)J sequencing and monoclonal antibody generation identified somatic hypermutation driven affinity maturation to SFB antigens under homeostatic conditions. Combining phage display and B cell tetramers will enable investigation of the ontogeny and function of commensal-specific B cell responses in tissue immunity, inflammation, and repair.
Pubmed ID: 38723638
Research resources used in this publication
- B6.129P2-Tcrdtm1Mom/J (RRID:IMSR_JAX:002120) (organism)
- AbVec2.0-IGHG1 (RRID:Addgene_80795) (plasmid)
- AbVec1.1-IGKC (RRID:Addgene_80796) (plasmid)
- Phangorn (RRID:SCR_017302) (software resource)
- shazam (RRID:SCR_024301) (software resource)
- Change-O (RRID:SCR_023986) (software resource)
- Seurat (RRID:SCR_016341) (software resource)
- Cell Ranger (RRID:SCR_017344) (software resource)
- SnapGene (RRID:SCR_015052) (annotation tool)
- Biorender (RRID:SCR_018361) (web application)
- FlowJo (RRID:SCR_008520) (software resource)
- Adobe Illustrator (RRID:SCR_010279) (software resource)
- ImageJ (RRID:SCR_003070) (software resource)
- GraphPad Prism (RRID:SCR_002798) (software resource)
- STOCK Tg(Cd4-cre)1Cwi/BfluJ (RRID:IMSR_JAX:017336) (organism)
- C57BL/6J (RRID:IMSR_JAX:000664) (organism)
- B6.129P2-Icostm1Mak/J (RRID:IMSR_JAX:004859) (organism)
- C57BL/6NTac (RRID:IMSR_TAC:B6) (organism)
- B6.129P2-Tcrbtm1Mom/J (RRID:IMSR_JAX:002118) (organism)
Additional research tools detected in this publication
None foundAntibodies used in this publication
- TotalSeq(TM)-C0310 anti-mouse Hashtag 10 (RRID:AB_2819914)
- TotalSeq(TM)-C0309 anti-mouse Hashtag 9 (RRID:AB_2819913)
- TotalSeq(TM)-C0308 anti-mouse Hashtag 8 (RRID:AB_2819912)
- TotalSeq(TM)-C0307 anti-mouse Hashtag 7 (RRID:AB_2819911)
- TotalSeq(TM)-C0306 anti-mouse Hashtag 6 (RRID:AB_2819910)
- TotalSeq(TM)-C0305 anti-mouse Hashtag 5 (RRID:AB_2800697)
- TotalSeq(TM)-C0304 anti-mouse Hashtag 4 (RRID:AB_2800696)
- TotalSeq(TM)-C0303 anti-mouse Hashtag 3 (RRID:AB_2800695)
- TotalSeq(TM)-C0302 anti-mouse Hashtag 2 (RRID:AB_2800694)
- TotalSeq(TM)-C0301 anti-mouse Hashtag 1 (RRID:AB_2800693)
- Goat Anti-Human IgG-HRP (RRID:AB_2795644)
- Alexa Fluor(R) 488 anti-mouse CD326 (Ep-CAM) (RRID:AB_1134099)
- Hu IgG R718 G18-145 50ug (RRID:AB_2917040)
- M13 Phage coat protein Polyclonal Antibody (RRID:AB_795743)
- Peroxidase-AffiniPure Goat Anti-Mouse IgG, Fc_ Subclass 3 Specific (min X Hu,Bov,Rb Sr Prot) (RRID:AB_2338517)
- AffiniPure Goat Anti-Mouse IgG, Fc_ Subclass 2c Specific (min X Hu,Bov,Rb Sr Prot) (RRID:AB_2338464)
- Goat Anti-Mouse IgG2b, Human ads-HRP (RRID:AB_2794521)
- Peroxidase-AffiniPure Goat Anti-Mouse IgM, µ Chain Specific (RRID:AB_2338502)
- Goat Anti-Mouse IgE-HRP (RRID:AB_2794604)
- IgA (RRID:AB_2314669)
- Goat Anti-Bovine Serum Albumin, Human Serum Albumin ads-UNLB (RRID:AB_2795778)
- ReadyTag anti-OVA (RRID:AB_2651130)
- CD45 Monoclonal Antibody (30-F11), APC-eFluor™ 780, eBioscience (RRID:AB_1548781)
- Goat Anti-Mouse IgG1, Human ads-HRP (RRID:AB_2650509)
- Brilliant Violet 785(TM) anti-mouse Ly-6G (RRID:AB_2566317)
- Goat Anti-Mouse IgA-HRP (RRID:AB_2714213)
- Brilliant Violet 785(TM) anti-mouse CD11c (RRID:AB_2565268)
- Brilliant Violet 785(TM) anti-mouse F4/80 (RRID:AB_2563667)
- Brilliant Violet 785(TM) anti-mouse CD90.2 (Thy1.2) (RRID:AB_2562900)
- Goat Anti-Mouse IgG, Human ads-UNLB (RRID:AB_2794290)
- PE/Cyanine7 anti-mouse IgG2b (RRID:AB_2750280)
- Alexa Fluor(R) 700 anti-mouse CD38 (RRID:AB_2890672)
- Goat Anti-Mouse IgA-FITC (RRID:AB_2794370)
- IgM (RRID:AB_2741428)
- Brilliant Violet 510(TM) anti-mouse IgD (RRID:AB_2562742)
- PerCP/Cyanine5.5 anti-mouse IgG1 (RRID:AB_2562000)
- Brilliant Violet 650(TM) anti-mouse CD138 (Syndecan-1) (RRID:AB_2650927)
- Pacific Blue(TM) anti-MU/HU GL7 Antigen (T/B Cell Act. Marker) (RRID:AB_2563291)
- Ms CD95 BUV805 Jo2 50ug (RRID:AB_2871273)
- Ms CD45R/B220 BUV737 RA3-6B2 50ug (RRID:AB_2870160)
Associated grants
- Agency: NIAID NIH HHS, United States
Id: R01 AI158624 - Agency: NIAID NIH HHS, United States
Id: R21 AI171921 - Agency: NIGMS NIH HHS, United States
Id: T32 GM136534
Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.
This is a list of tools and resources that we have found mentioned in this publication.
TotalSeq(TM)-C0310 anti-mouse Hashtag 10 (antibody)
RRID:AB_2819914
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0309 anti-mouse Hashtag 9 (antibody)
RRID:AB_2819913
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0308 anti-mouse Hashtag 8 (antibody)
RRID:AB_2819912
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0307 anti-mouse Hashtag 7 (antibody)
RRID:AB_2819911
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0306 anti-mouse Hashtag 6 (antibody)
RRID:AB_2819910
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0305 anti-mouse Hashtag 5 (antibody)
RRID:AB_2800697
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0304 anti-mouse Hashtag 4 (antibody)
RRID:AB_2800696
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0303 anti-mouse Hashtag 3 (antibody)
RRID:AB_2800695
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0302 anti-mouse Hashtag 2 (antibody)
RRID:AB_2800694
This monoclonal targets H-2/CD45
View all literature mentionsTotalSeq(TM)-C0301 anti-mouse Hashtag 1 (antibody)
RRID:AB_2800693
This monoclonal targets H-2/CD45
View all literature mentionsGoat Anti-Human IgG-HRP (antibody)
RRID:AB_2795644
This unknown targets IgG
View all literature mentionsAlexa Fluor(R) 488 anti-mouse CD326 (Ep-CAM) (antibody)
RRID:AB_1134099
This monoclonal targets CD326
View all literature mentionsHu IgG R718 G18-145 50ug (antibody)
RRID:AB_2917040
This unknown targets IgG
View all literature mentionsM13 Phage coat protein Polyclonal Antibody (antibody)
RRID:AB_795743
This unknown targets M13 Phage coat protein
View all literature mentionsPeroxidase-AffiniPure Goat Anti-Mouse IgG, Fc_ Subclass 3 Specific (min X Hu,Bov,Rb Sr Prot) (antibody)
RRID:AB_2338517
This unknown targets Mouse IgG, Fc? Subclass 3 Specific
View all literature mentionsAffiniPure Goat Anti-Mouse IgG, Fc_ Subclass 2c Specific (min X Hu,Bov,Rb Sr Prot) (antibody)
RRID:AB_2338464
This unknown targets Mouse IgG, Fc? Subclass 2c Specific
View all literature mentionsGoat Anti-Mouse IgG2b, Human ads-HRP (antibody)
RRID:AB_2794521
This unknown targets IgG2b
View all literature mentionsPeroxidase-AffiniPure Goat Anti-Mouse IgM, µ Chain Specific (antibody)
RRID:AB_2338502
This unknown targets Mouse IgM,
View all literature mentionsGoat Anti-Mouse IgE-HRP (antibody)
RRID:AB_2794604
This unknown targets IgE
View all literature mentionsGoat Anti-Bovine Serum Albumin, Human Serum Albumin ads-UNLB (antibody)
RRID:AB_2795778
This unknown targets Serum Albumin
View all literature mentionsReadyTag anti-OVA (antibody)
RRID:AB_2651130
This monoclonal targets OVA
View all literature mentionsCD45 Monoclonal Antibody (30-F11), APC-eFluor™ 780, eBioscience (antibody)
RRID:AB_1548781
This monoclonal targets CD45
View all literature mentionsGoat Anti-Mouse IgG1, Human ads-HRP (antibody)
RRID:AB_2650509
This polyclonal targets heavy chain of mouse IgG1
View all literature mentionsBrilliant Violet 785(TM) anti-mouse Ly-6G (antibody)
RRID:AB_2566317
This monoclonal targets Ly-6G
View all literature mentionsGoat Anti-Mouse IgA-HRP (antibody)
RRID:AB_2714213
This polyclonal targets Heavy chain of mouse IgA
View all literature mentionsBrilliant Violet 785(TM) anti-mouse CD11c (antibody)
RRID:AB_2565268
This monoclonal targets CD11c
View all literature mentionsBrilliant Violet 785(TM) anti-mouse F4/80 (antibody)
RRID:AB_2563667
This monoclonal targets F4/80
View all literature mentionsBrilliant Violet 785(TM) anti-mouse CD90.2 (Thy1.2) (antibody)
RRID:AB_2562900
This monoclonal targets CD90.2
View all literature mentionsGoat Anti-Mouse IgG, Human ads-UNLB (antibody)
RRID:AB_2794290
This unknown targets IgG
View all literature mentionsPE/Cyanine7 anti-mouse IgG2b (antibody)
RRID:AB_2750280
This monoclonal targets IgG2b
View all literature mentionsAlexa Fluor(R) 700 anti-mouse CD38 (antibody)
RRID:AB_2890672
This monoclonal targets CD38
View all literature mentionsGoat Anti-Mouse IgA-FITC (antibody)
RRID:AB_2794370
This unknown targets IgA
View all literature mentionsBrilliant Violet 510(TM) anti-mouse IgD (antibody)
RRID:AB_2562742
This monoclonal targets IgD
View all literature mentionsPerCP/Cyanine5.5 anti-mouse IgG1 (antibody)
RRID:AB_2562000
This monoclonal targets IgG1
View all literature mentionsBrilliant Violet 650(TM) anti-mouse CD138 (Syndecan-1) (antibody)
RRID:AB_2650927
This monoclonal targets CD138
View all literature mentionsPacific Blue(TM) anti-MU/HU GL7 Antigen (T/B Cell Act. Marker) (antibody)
RRID:AB_2563291
This monoclonal targets GL7
View all literature mentionsMs CD95 BUV805 Jo2 50ug (antibody)
RRID:AB_2871273
This monoclonal targets CD95 (Fas)
View all literature mentionsMs CD45R/B220 BUV737 RA3-6B2 50ug (antibody)
RRID:AB_2870160
This monoclonal targets CD45R (B220)
View all literature mentionsB6.129P2-Tcrdtm1Mom/J (organism)
RRID:IMSR_JAX:002120
Mus musculus with name B6.129P2-Tcrdtm1Mom/J from IMSR.
View all literature mentionsPhangorn (software resource)
RRID:SCR_017302
Software R package for phylogenetic reconstruction and analysis. Used for estimation of phylogenetic trees and networks using Maximum Likelihood, Maximum Parsimony, distance methods and Hadamard conjugation. Allows to compare trees, models selection and offers visualizations for trees and split networks.
View all literature mentionsshazam (software resource)
RRID:SCR_024301
Software R package provides computational framework for analyzing mutations in immunoglobulin sequences. Immunoglobulin Somatic Hypermutation Analysis.
View all literature mentionsChange-O (software resource)
RRID:SCR_023986
Collection of software tools for processing the output of V(D)J alignment tools, assigning clonal clusters to immunoglobulin Ig sequences, and reconstructing germline sequences.
View all literature mentionsSeurat (software resource)
RRID:SCR_016341
Software as R package designed for QC, analysis, and exploration of single cell RNA-seq data. Enable users to identify and interpret sources of heterogeneity from single cell transcriptomic measurements, and to integrate diverse types of single cell data.
View all literature mentionsCell Ranger (software resource)
RRID:SCR_017344
Software tool as set of analysis pipelines that process chromium single cell RNA-seq output to align reads, generate feature-barcode matrices and perform clustering and gene expression analysis by 10xGenomics.
View all literature mentionsSnapGene (annotation tool)
RRID:SCR_015052
Molecular biology software for visualizing and documenting gene constructs for InFusion cloning, Gibson assembly, restriction cloning, PCR, and mutagenesis.
View all literature mentionsBiorender (web application)
RRID:SCR_018361
Web tool for graphical illustrations for biological systems by BioRender. Used for articles and presentations. Collection of pre made icons and templates from life science fields for scientific illustrations. Customized icons possible on request.
View all literature mentionsFlowJo (software resource)
RRID:SCR_008520
Software for single-cell flow cytometry analysis. Its functions include management, display, manipulation, analysis and publication of the data stream produced by flow and mass cytometers.
View all literature mentionsAdobe Illustrator (software resource)
RRID:SCR_010279
Vector graphics software to create digital graphics, illustrations, and typography for several types of media: print, web, interactive, video, and mobile.
View all literature mentionsImageJ (software resource)
RRID:SCR_003070
Open source Java based image processing software program designed for scientific multidimensional images. ImageJ has been transformed to ImageJ2 application to improve data engine to be sufficient to analyze modern datasets.
View all literature mentionsGraphPad Prism (software resource)
RRID:SCR_002798
Statistical analysis software that combines scientific graphing, comprehensive curve fitting (nonlinear regression), understandable statistics, and data organization. Designed for biological research applications in pharmacology, physiology, and other biological fields for data analysis, hypothesis testing, and modeling.
View all literature mentionsSTOCK Tg(Cd4-cre)1Cwi/BfluJ (organism)
RRID:IMSR_JAX:017336
Mus musculus with name STOCK Tg(Cd4-cre)1Cwi/BfluJ from IMSR.
View all literature mentionsC57BL/6J (organism)
RRID:IMSR_JAX:000664
Mus musculus with name C57BL/6J from IMSR.
View all literature mentionsB6.129P2-Icostm1Mak/J (organism)
RRID:IMSR_JAX:004859
Mus musculus with name B6.129P2-Icostm1Mak/J from IMSR.
View all literature mentionsC57BL/6NTac (organism)
RRID:IMSR_TAC:B6
Mus musculus with name C57BL/6NTac from IMSR.
View all literature mentionsB6.129P2-Tcrbtm1Mom/J (organism)
RRID:IMSR_JAX:002118
Mus musculus with name B6.129P2-Tcrbtm1Mom/J from IMSR.
View all literature mentions
