Suppression of cell proliferation and cytokine expression by HL-p36, a tick salivary gland-derived protein of Haemaphysalis longicornis.
Previously, a putative immunosuppressant-coding gene was identified from a complementary DNA library derived from the salivary glands of partially-fed Haemaphysalis longicornis. Using real-time polymerase chain reaction, the gene was shown to be predominantly expressed during blood feeding with the site of expression being mainly in the salivary glands; this was confirmed by Western blotting analysis. To investigate the function of this novel protein, in this study, we examined the proliferative responses of bovine mononuclear cells and murine splenic cells as well as the expression of profiles of several cytokines in these cells in the presence of the recombinant protein (H. longicornis-derived 36 000 molecular weight protein: rHL-p36). The addition of rHL-p36 at the beginning of the 72 hr cultivation period clearly inhibited proliferation of several mitogen-stimulated cells in a dose-dependent manner, with concomitantly significant down-regulation of messenger RNA levels for interleukin-2. The inhibitory response could be abrogated by blockage of HL-p36 with antibody, suggesting the direct involvement of rHL-p36 in the cell proliferation. Furthermore, the proliferative response of splenocytes isolated from rHL-p36-inoculated mice was significantly lower than for those from control mice, suggesting that rHL-p36 could also directly suppress immune responses in vivo. Interestingly, microarray analysis of the splenocytes showed that the expression of several immunomodulating genes was down-regulated by rHL-p36 inoculation. In conclusion, these results suggest that HL-p36 is an immunosuppressor that might play an important role in the modulation of host immune responses.
Konnai
Satoru
S
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan. konnai@vetmed.hokudai.ac.jp
Nakajima
Chie
C
Imamura
Saiki
S
Yamada
Shinji
S
Nishikado
Hideto
H
Kodama
Michi
M
Onuma
Misao
M
Ohashi
Kazuhiko
K
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D049109:Cell Proliferation; D016207:Cytokines / Q000096:biosynthesis*; D018076:DNA, Complementary / Q000235:genetics; D004306:Dose-Response Relationship, Immunologic; D005786:Gene Expression Regulation / Q000276:immunology; D007108:Immune Tolerance / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012469:Salivary Glands / Q000276:immunology*; D012471:Salivary Proteins and Peptides / Q000235:genetics / Q000276:immunology*; D017422:Sequence Analysis, DNA / Q000379:methods; D013154:Spleen / Q000276:immunology; D013987:Ticks / Q000276:immunology*
D016207:Cytokines; D018076:DNA, Complementary; C539747:HL-p36 protein, Haemaphysalis longicornis; D012333:RNA, Messenger; D011994:Recombinant Proteins; D012471:Salivary Proteins and Peptides