Comparison of phage pVIII and KLH as vector in inducing the production of cytokines in C57BL/6J mice.
We have demonstrated that phage display Candida albicans (C. albicans) LKVIRK epitope was protective in systemically infected C57BL/6J mice. The different development from precursor Ths, Th1 or Th2, will result in a protective or nonprotective immune response. To compare the types of cytokines induced by biologically and chemically synthesized vectors, C57BL/6J mice were immunized with hybrid phage displaying the epitope of LKVIRK and by synthesized peptide epitope LKVIRKNIVKKMIE conjugated through cysteine to keyhole limpet haemocyanin (KLH). The production of cytokines in spleens of immunized mice and in splenocytes culture supernatants stimulated by homologous immunogen in vitro was studied by RT-PCR and quantitative sandwich ELISA. The results showed that, compared to Tris-EDTA buffer (TE, 1 mM Tris, 0.1 mM EDTA, pH 8.0) injected mice, the expressions of Th1 type cytokine IFN-gamma, IL-2 and IL-12 were increased in hybrid phage, KLH-C, and wild phage immunized mice, and there were no differences between mice immunized with hybrid phage and KLH-C. While the expression of Th2 type cytokine IL-10 was similar in all mice, IL-4 was not detected. We obtained the same results in mRNA and protein level. These findings indicated that as carriers, phage and KLH were similar in inducing the Th1 type cytokines expression. Comparing to peptide synthesis couple with a carrier protein for injection, phage may be an inexpensive and simple route to the production of effective vaccines.
Su
Quan-Ping
QP
Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, 5268 Renmin Street, Changchun City, Jilin Province 130024, People's Republic of China.
Wen
De-Zhong
DZ
Yang
Qiong
Q
Zhang
Yan-Hui
YH
Liu
Chong
C
Wang
Li
L
D000818:Animals; D001435:Bacteriophages / Q000235:genetics* / Q000276:immunology; D002176:Candida albicans / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013154:Spleen / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology
D036022:Capsid Proteins; D016207:Cytokines; D000939:Epitopes; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin