title: Primary anti-trinitrophenyl memory cells investigated by plaque-forming cells and ELISA.
abstract: Primary anti-trinitrophenyl antibody production was investigated from spleen cells of mice immunized with trinitrophenylated-keyhole limpet hemocyanin, using the plaque-forming cell method and ELISA. Cells taken 5 days after antigen injection do not produce IgE, but do produce IgM and IgG1 anti-trinitrophenyl antibodies as demonstrated by plaque-forming cells. Substantial increase of IgM, IgG1, and IgE antibody production was seen from cells taken 7 days after immunization, followed by a rapid decline. By ELISA it was seen that cells taken 3 days after immunization already produce small amounts of anti-trinitrophenyl antibodies. Presence of antigen from the start of the cultures did not increase antibody production from cells taken 3 days after immunization, but potentiated antibody secretions from cells taken 5 days or later after immunization. This potentiation was interpreted as recruitment of antibody-forming cells from early memory B cells. The presence of IL-4 from the start of the cultures had no appreciable effect. Cell sorting with specific antibody-coated magnetic beads showed that plaque-forming cells from nonsorted cells, membrane IgE+ or membrane IgE- cells secreted similar amounts of anti-trinitrophenyl IgG1 and IgE antibodies. No difference in anti-trinitrophenyl IgM, IgG1, or IgE production was found in controls; cells sorted negatively or positively for CD23. The data show that memory B cells can be demonstrated already on Day 5 after immunization, and their antigen-induced antibody secretion is IL-4 dependent.
authors.affiliation: Department of Pathology, New York University Medical School, New York 10016.
authors.forename: A
authors.initials: A
authors.lastname: Ueda
authors.forename: J
authors.initials: J
authors.lastname: Leu
authors.forename: Z
authors.initials: Z
authors.lastname: Ovary
mesh_terms: D000818:Animals; D000917:Antibody Formation; D000921:Antibody-Producing Cells / Q000276:immunology*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D016175:B-Lymphocyte Subsets / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens; D007073:Immunoglobulin E / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D007156:Immunologic Memory; D015847:Interleukin-4 / Q000502:physiology; D051379:Mice; D008815:Mice, Inbred Strains; D011947:Receptors, Antigen, B-Cell / Q000276:immunology; D011961:Receptors, Fc / Q000032:analysis; D017455:Receptors, IgE; D014301:Trinitrobenzenes / Q000276:immunology*
chemical_list: D000944:Antigens, Differentiation, B-Lymphocyte; D006241:Haptens; D007074:Immunoglobulin G; D011947:Receptors, Antigen, B-Cell; D011961:Receptors, Fc; D017455:Receptors, IgE; D014301:Trinitrobenzenes; D015847:Interleukin-4; D007073:Immunoglobulin E