Germinal center expansion but not plasmablast differentiation is proportional to peptide-MHCII density via CD40-CD40L signaling strength.
T follicular helper (TFH) cells promote expansion of germinal center (GC) B cells and plasma cell differentiation. Whether cognate peptide-MHCII (pMHCII) density instructs selection and cell fate decisions in a quantitative manner remains unclear. Using αDEC205-OVA to differentially deliver OVA peptides to GC B cells on the basis of DEC205 allelic copy number, we find DEC205+/+ B cells take up 2-fold more antigen than DEC205+/- cells, leading to proportional TFH cell help and B cell expansion. To validate these results, we establish a caged OVA peptide, which is readily detected by OVA-specific TFH cells after photo-uncaging. In situ uncaging of peptides leads to multiple serial B-T contacts and cell activation. Differential CD40 signaling, is both necessary and sufficient to mediate 2-fold differences in B cell expansion. While plasmablast numbers are increased, pMHCII density does not directly control the output or quality of plasma cells. Thus, we distinguish the roles TFH cells play in expansion versus differentiation.
Pubmed ID: 35508132 RIS Download
Research resources used in this publication
- B6.Cg-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J (RRID:IMSR_JAX:007914) (organism)
- B6.Cg-Gt(ROSA)26Sortm2(CAG-PA-GFP)Rmpl/J (RRID:IMSR_JAX:035687) (organism)
- B6.C-Tg(CMV-cre)1Cgn/J (RRID:IMSR_JAX:006054) (organism)
- B6.Cg-Tg(TcraTcrb)425Cbn/J (RRID:IMSR_JAX:004194) (organism)
- B6.Cg-Tg(Prdm1-EYFP)1Mnz/J (RRID:IMSR_JAX:008828) (organism)
- STOCK Tg(CAG-ECFP)CK6Nagy/J (RRID:IMSR_JAX:003773) (organism)
-
B6.SJL-PtprcaPepc
- Imaris (RRID:SCR_007370) (software resource)
- CBy.129P2(B6)-Ightm1Mnz/J (RRID:IMSR_JAX:007775) (organism)
- Microsoft Excel (RRID:SCR_016137) (software resource)
- Volocity 3D Image Analysis Software (RRID:SCR_002668) (software resource)
- GraphPad Prism (RRID:SCR_002798) (software resource)
- FlowJo (RRID:SCR_008520) (software resource)
- Adobe Illustrator (RRID:SCR_010279) (software resource)
- HEK293T (RRID:CVCL_0063) (cell line)
Additional research tools detected in this publication
Antibodies used in this publication
- Alexa Fluor(R) 647 anti-mouse TCR Valpha2 (RRID:AB_1186115)
- Rabbit Anti-S6 Ribosomal Protein, phospho (Ser235 / Ser236) Monoclonal Antibody, Alexa Fluor?? 647 Conjugated, Clone D57.2.2E (RRID:AB_10695457)
- IgMa (RRID:AB_394897)
- PerCP/Cyanine5.5 anti-MU/HU GL7 Antigen (T/B Cell Act. Marker) (RRID:AB_2562979)
- T- and B-Cell Activation Antigen (RRID:AB_10716056)
- Purified anti-mouse CD205 (DEC-205) (RRID:AB_2281398)
- Rat Anti-Mouse CD138 Monoclonal Antibody, APC Conjugated, Clone 281-2 (RRID:AB_1645216)
- CD86 (B7-2) (RRID:AB_2738663)
- Mouse Anti-Mouse CD45.2 Monoclonal Antibody, V450 Conjugated, Clone 104 (RRID:AB_1727495)
- Mouse Anti-CD45.2 Monoclonal Antibody, PerCP-Cy5.5 Conjugated, Clone 104 (RRID:AB_394528)
- CD95 (Fas) (RRID:AB_2738345)
- CD69 Monoclonal Antibody (H1.2F3), FITC, eBioscience (RRID:AB_465119)
- InVivoPlus anti-mouse CD154 (CD40L) (RRID:AB_1107601)
- IRF8 Monoclonal Antibody (V3GYWCH), APC, eBioscience (RRID:AB_2573318)
- IRF4 Monoclonal Antibody (3E4), PE, eBioscience (RRID:AB_10852721)
- Rat IgG2a kappa Isotype Control (eBR2a), Functional Grade, eBioscience (RRID:AB_470156)
- PE anti-mouse IgMa (RRID:AB_940545)
- APC anti-mouse CD205 (DEC-205) (RRID:AB_10613641)
- CD184 (RRID:AB_394305)
- PerCP/Cyanine5.5 anti-mouse CD138 (Syndecan-1) (RRID:AB_2561601)
- Rat Anti-CD138 Monoclonal Antibody, Phycoerythrin Conjugated, Clone 281-2 (RRID:AB_395000)
- PerCP/Cyanine5.5 anti-mouse CD45.2 (RRID:AB_893350)
- CD45.1 Monoclonal Antibody (A20), PE, eBioscience (RRID:AB_465676)
- CD45.1 Monoclonal Antibody (A20), eFluor™ 450, eBioscience (RRID:AB_1272189)
- CD45.1 (RRID:AB_10611577)
- CD40 Monoclonal Antibody (1C10), Functional Grade, eBioscience (RRID:AB_468941)
- CD38 (RRID:AB_2737782)
- anti-mouse CD16/32 Antibody (RRID:AB_2687830)
- Brilliant Violet 421(TM) anti-mouse CD4 (RRID:AB_11203718)
- APC/Cyanine7 anti-mouse/human CD45R/B220 (RRID:AB_313007)
- Ea52-68 peptide bound to I-Ab Monoclonal Antibody (eBioY-Ae (YAe, Y-Ae)), Biotin, eBioscience (RRID:AB_657821)
- InVivoPlus polyclonal Armenian hamster IgG (RRID:AB_1107773)
- PE-Cy™7 Hamster Anti-Mouse CD95 (RRID:AB_396768)
Associated grants
- Agency: NHLBI NIH HHS, United States
Id: R01 HL141491 - Agency: NCI NIH HHS, United States
Id: P30 CA013330 - Agency: NIAID NIH HHS, United States
Id: R01 AI072529 - Agency: NCRR NIH HHS, United States
Id: S10 RR023704
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This is a list of tools and resources that we have found mentioned in this publication.
GENEWIZ (tool)
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RRID:SCR_007370
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RRID:SCR_002798
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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.
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Vector graphics software to create digital graphics, illustrations, and typography for several types of media: print, web, interactive, video, and mobile.
View all literature mentionsHEK293T (cell line)
RRID:CVCL_0063
Cell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)
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