The secondary antigen-specific IgE response in murine lymphocytes is resistant to blockade by anti-IL4 antibody and an antisense oligodeoxynucleotide for IL4 mRNA.
In order to investigate the role of interleukin 4 (IL4) in the induction of antigen-specific IgE responses, we established culture conditions which allow the induction of anti-trinitrophenyl(TNP) IgE response by the coculture of TNP-keyhole limpet hemocyanin-primed C3H B cells with conalbumin (CA)-specific type 2 helper T (Th2) cell clone, D10.G4.1 in the presence of TNP-CA. A maximum level of anti-TNP IgE was secreted at 1 microgram/ml TNP-CA. By using filter-separated double-chamber culture plates, the physical contact between T cells and B cells was shown to be necessary in this response. Anti-TNP IgE synthesis was not significantly suppressed in the presence of 20-40 micrograms/ml monoclonal anti-IL4 (11B11), nor was the response further enhanced by the addition of recombinant IL4. 11B11 added together with anti-IL5 had also no suppressive effects on the IgE response. This apparent independence on IL4 might be due to the fact that IL4 is transferred from T cells to B cells in a transsynaptical way that would be refractory to the neutralization by 11B11. In order to test this possibility, we synthesized the phosphorothioate analogue of an antisense oligodeoxynucleotide against IL4 mRNA (S-oligo) for inhibiting IL4 production from Th2 cells specifically. S-oligo was effective at 10-20 micrograms/ml in suppressing IL4 production from D10.G4.1 cells by 80-90%. It was demonstrated that S-oligo, either alone or in combination with 11B11, did not significantly suppress anti-TNP IgE response. These results suggest that antigen-specific IgE response in primed B cells does not depend on IL4, but requires cognate interaction with Th2 cells.
Haruna
K
K
Department of Biotechnology, Faculty of Engineering, Okayama University, Japan.
Hikida
M
M
Ohsugi
Y
Y
Ohmori
H
H
D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology*; D001483:Base Sequence; D005260:Female; D007073:Immunoglobulin E / Q000096:biosynthesis*; D015847:Interleukin-4 / Q000096:biosynthesis / Q000502:physiology*; D008211:Lymphocyte Cooperation; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger / Q000235:genetics; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D014301:Trinitrobenzenes / Q000276:immunology
D000906:Antibodies; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D014301:Trinitrobenzenes; D015847:Interleukin-4; D007073:Immunoglobulin E