Identification and characterization of the constituent human serum antibodies elicited by vaccination.
Most vaccines confer protection via the elicitation of serum antibodies, yet more than 100 y after the discovery of antibodies, the molecular composition of the human serum antibody repertoire to an antigen remains unknown. Using high-resolution liquid chromatography tandem MS proteomic analyses of serum antibodies coupled with next-generation sequencing of the V gene repertoire in peripheral B cells, we have delineated the human serum IgG and B-cell receptor repertoires following tetanus toxoid (TT) booster vaccination. We show that the TT(+) serum IgG repertoire comprises ∼100 antibody clonotypes, with three clonotypes accounting for >40% of the response. All 13 recombinant IgGs examined bound to vaccine antigen with Kd ∼ 10(-8)-10(-10) M. Five of 13 IgGs recognized the same linear epitope on TT, occluding the binding site used by the toxin for cell entry, suggesting a possible explanation for the mechanism of protection conferred by the vaccine. Importantly, only a small fraction (<5%) of peripheral blood plasmablast clonotypes (CD3(-)CD14(-)CD19(+)CD27(++)CD38(++)CD20(-)TT(+)) at the peak of the response (day 7), and an even smaller fraction of memory B cells, were found to encode antibodies that could be detected in the serological memory response 9 mo postvaccination. This suggests that only a small fraction of responding peripheral B cells give rise to the bone marrow long-lived plasma cells responsible for the production of biologically relevant amounts of vaccine-specific antibodies (near or above the Kd). Collectively, our results reveal the nature and dynamics of the serological response to vaccination with direct implications for vaccine design and evaluation.
Department of Chemical Engineering, Institute for Cellular and Molecular Biology, Section of Molecular Genetics and Microbiology, Center for Systems and Synthetic Biology, Department of Biomedical Engineering, Department of Chemistry and Biochemistry, and Applied Research Laboratories, University of Texas at Austin, Austin, TX 78712-1062.
Jason J
JJ
Lavinder
Yariv
Y
Wine
Claudia
C
Giesecke
Gregory C
GC
Ippolito
Andrew P
AP
Horton
Oana I
OI
Lungu
Kam Hon
KH
Hoi
Brandon J
BJ
DeKosky
Ellen M
EM
Murrin
Megan M
MM
Wirth
Andrew D
AD
Ellington
Thomas
T
Dörner
Edward M
EM
Marcotte
Daniel R
DR
Boutz
George
G
Georgiou
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000096:biosynthesis* / Q000737:chemistry; D001402:B-Lymphocytes / Q000276:immunology; D002853:Chromatography, Liquid; D006801:Humans; D016130:Immunophenotyping; D008969:Molecular Sequence Data; D053719:Tandem Mass Spectrometry; D013745:Tetanus Toxoid / Q000008:administration & dosage*
D000907:Antibodies, Bacterial; D013745:Tetanus Toxoid
B-cell repertoire; proteomics