In vitro induction of a Haemophilus influenzae type b polysaccharide-specific antibody response in human peripheral blood lymphocytes of individuals recently vaccinated with an oligosaccharide-protein conjugate.
In this paper an in vitro culture system for the induction of an antibody response to the Haemophilus influenzae type b polysaccharide (PRP) is described. Anti-PRP IgM and IgG antibody-secreting cells (ASC) and anti-diphtheria toxoid (DT) IgG ASC were detected in cultures of blood B and T cells derived from donors 4 to 6 wk after immunization with Haemophilus influenzae type b oligosaccharide-mutant diphtheria toxin (CRM197) conjugate (HbOC) and required in vitro restimulation with HbOC. When lymphocytes from HbOC-vaccinated donors were stimulated with PRP, anti-PRP IgM and IgG ASC could be detected in 50% offGe cases. Lymphocytes from PRP-vaccinated donors or non-vaccinated donors consistently failed to generate anti-PRP antibodies after in vitro stimulation with HbOC. Optimal in vitro responses were observed at concentrations of 0.06 to 0.6 micrograms/ml of Ag. At higher doses of Ag (6 micrograms/ml) anti-PRP and anti-DT antibody responses were suppressed. The in vitro generation of anti-PRP and anti-DT ASC, as detected by a spot-forming cell assay was shown to be T cell dependent, Ag dependent, and Ag specific. This culture system provides a model for the study of human B cell activation and immunoregulation by polysaccharide-protein conjugates and polysaccharides.
Peeters
C C
CC
Department of Immunology, University Hospital for Children and Youth het Wilhelmina Kinderziekenhuis, Utrecht, The Netherlands.
Tenbergen-Meekes
A M
AM
Heijnen
C J
CJ
Poolman
J T
JT
Zegers
B J
BJ
Rijkers
G T
GT
D000328:Adult; D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D016667:Bacterial Capsules; D001428:Bacterial Vaccines / Q000276:immunology*; D004167:Diphtheria Toxin / Q000276:immunology; D004168:Diphtheria Toxoid / Q000276:immunology*; D018073:Haemophilus Vaccines; D006193:Haemophilus influenzae / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000276:immunology*; D011135:Polysaccharides, Bacterial / Q000276:immunology; D014611:Vaccination
D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D004167:Diphtheria Toxin; D004168:Diphtheria Toxoid; D018073:Haemophilus Vaccines; C044237:Haemophilus influenzae type b polysaccharide vaccine; C052180:Haemophilus influenzae type b-polysaccharide vaccine-diphtheria toxoid conjugate; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011135:Polysaccharides, Bacterial