Preparation and analysis of antigen-specific memory B cells.
A procedure has been developed for the enrichment of TNP-binding memory B cells (TNP-MABC) from spleens of immunized mice. More than 75% of the cells expressed surface IgM (sIgM) and IgD (sIgD) and about 9% expressed surface IgG (sIgG). The TNP-MABC consisted of small resting lymphocytes with high affinity antigen-binding receptors. These cells expressed increased densities of Ia antigens and decreased densities of sIgD. Adoptive transfer of the cells into irradiated, carrier-primed syngeneic recipients resulted in their differentiation into IgG anti-TNP antibody-secreting cells. TNP-MABC secreted high affinity IgG anti-TNP antibodies when cultured in vitro with carrier-primed T cells and antigen. Limiting dilution analysis revealed that TNP-MABC contained a relatively low frequency of precursors for IgG-secreting cells that had an exceptionally large clone size. These results show that a highly enriched population of antigen-specific memory B cells can now be prepared and used to analyze their activation requirements.
Yefenof
E
E
Sanders
V M
VM
Snow
E C
EC
Noelle
R J
RJ
Oliver
K G
KG
Uhr
J W
JW
Vitetta
E S
ES
D000818:Animals; D000915:Antibody Affinity; D001402:B-Lymphocytes / Q000145:classification* / Q000166:cytology / Q000378:metabolism / Q000637:transplantation; D002453:Cell Cycle; D002469:Cell Separation / Q000379:methods*; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006462:Hemolytic Plaque Technique; D000949:Histocompatibility Antigens Class II / Q000032:analysis; D007116:Immunization, Passive; D007156:Immunologic Memory; D007958:Leukocyte Count; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D010641:Phenotype; D011946:Receptors, Antigen / Q000032:analysis; D011947:Receptors, Antigen, B-Cell / Q000032:analysis; D012397:Rosette Formation; D013234:Stem Cells / Q000276:immunology / Q000378:metabolism; D014301:Trinitrobenzenes / Q000276:immunology / Q000378:metabolism
D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D011946:Receptors, Antigen; D011947:Receptors, Antigen, B-Cell; D014301:Trinitrobenzenes