Adjuvant effect of Bacillus firmus on the expression of cytokines and toll-like receptors in mouse nasopharynx-associated lymphoid tissue (NALT) after intranasal immunization with inactivated influenza virus type A.
Due to the persisting threat of development of new highly pathogenic influenza A subtypes, a mucosal vaccination which would induce a potent and cross-protective reaction is desirable. We succeeded in mucosal immunization of mice with an inactivated influenza A virus by using delipidated Bacillus firmus (DBF) as adjuvant. The mechanism of adjuvant effect was followed in NALT by comparing the response after intranasal immunization by inactivated influenza virus type A (H1N1) alone, adjuvant alone (DBF), or by a mixture of virus+DBF. Expression of selected gene groups was tested via qPCR at 7 different time-points: cytokines (IL-2, IFN-γ, IL-4, IL-6, and IL-10), type I interferons (IFN-α4, IFN-α11, IFN-α12, and IFN-β), toll-like receptors (TLR2, TLR3, TLR7, and TLR9), iNOS and CCR7. Intranasally administered DBF and the mixture of virus+DBF induced an elevated expression of IFN-γ, IL-6 and IL-10 cytokines, type I interferons, iNOS, and pDC markers in NALT. Multimarker qPCR data was analyzed by relative quantification and by principal component analysis. DBF has been shown to be a very efficient adjuvant for the stimulation of innate immunity after IN immunization. DBF accelerated, increased, and prolonged the antiviral response.
Zanvit
Peter
P
Charles University in Prague, First Faculty of Medicine, Institute of Immunology and Microbiology, Studničkova 7, 128 00 Prague, Czech Republic. peter.zanvit@lf1.cuni.cz
Tichopád
Aleš
A
Havlíčková
Martina
M
Novotná
Olga
O
Jirkovská
Marie
M
Kološtová
Katarína
K
Cechová
Dana
D
Julák
Jaroslav
J
Sterzl
Ivan
I
Prokešová
Ludmila
L
D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000281:Administration, Intranasal; D000818:Animals; D001407:Bacillus / Q000276:immunology*; D016207:Cytokines / Q000235:genetics*; D004357:Drug Synergism; D015870:Gene Expression / Q000187:drug effects; D007114:Immunization / Q000379:methods; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000276:immunology; D007370:Interferon Type I / Q000235:genetics; D016753:Interleukin-10 / Q000235:genetics; D007376:Interleukin-2 / Q000235:genetics; D008221:Lymphoid Tissue / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D009305:Nasopharynx / Q000378:metabolism*; D025341:Principal Component Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D051195:Toll-Like Receptor 2 / Q000235:genetics; D051196:Toll-Like Receptor 3 / Q000235:genetics; D051199:Toll-Like Receptor 7 / Q000235:genetics; D051217:Toll-Like Receptor 9 / Q000235:genetics; D051193:Toll-Like Receptors / Q000235:genetics*
D000276:Adjuvants, Immunologic; D016207:Cytokines; D007252:Influenza Vaccines; D007370:Interferon Type I; D007376:Interleukin-2; D008562:Membrane Glycoproteins; C519703:TLR3 protein, mouse; C493486:Tlr2 protein, mouse; C477038:Tlr7 protein, mouse; D051195:Toll-Like Receptor 2; D051196:Toll-Like Receptor 3; D051199:Toll-Like Receptor 7; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors; D016753:Interleukin-10