Role as adhesin of muramidase released protein of Streptococcus s uis type 2

Size: px

Start display at page:

Download "Role as adhesin of muramidase released protein of Streptococcus s uis type 2"

Eleanore George
5 years ago
Views:

1 2002, 25 (4) : 6771 Journal of Nanjing Agricultural University 2 1,2 1 3, (11, ; 21, ) : 2 (SS2) (MRP) : 11 HA9801 (MRP + ) SH (MRP - ) HEp 2, MRP + MRP + ( P < 0105) h ; DNase HEp 2 HA MRP HEp 2, MRP MRP, MRP + ( P < 0105) ; MRP -, MRP MRP HA9801 : MRP SS2 : 2 ; ; ; : S : A : (2002) Role as adhesin of muramidase released protein of Streptococcus s uis type 2 ZENG Qiao2ying 1,2, LU Cheng2ping 1 3 (11Key Laboratory of Animal Disease Diagnostic and Immunology, Ministry of Agriculture, Nanjing Agric Univ, Nanjing , China ; 21Dept of Animal Medicine, Gansu Agric Univ, Lanzhou , China) Abstract : The adhesion of muramidase released protein (MRP), a virulence factor of Streptococcus suis type 2 (SS2) were tested. adhesive kinetics of SS2 isolate HA9801 (MRP + ) and SH (MRP - ) were compared through adhesive counting. Both strains could adhere to HEp22 cells, but the adhesive levels of MRP + strain were significantly higher than that of MRP - strain ( P < 0105). The ad2 hesive activities of HA9801 were abolished by pretreatment of bacterium cell with trypsin or galactose, or at 56 1 h, and by pretreat2 ment of HEp22 cells with periodic acid. After pretreatment of HEp22 cells with colchicine, lysozyme or DNase, the adhesive levels could be reduced dramatically. The adhesive activities of MRP + strain were obviously reduced but not blocked by pretreatment of bacterum cells with antiserum or monoclonal antibodies against MRP, or against MRP - The strain, as well as by pretreatment of HEp22 cells with puri2 fied MRP. MRP was destroyed completely its inhibitive performance against adhesion of HA9801 to HEp22 cells after pretreatment at 56 or with galactose, other than with trypsin, mercaptoethanol and hydrogen peroxide. Conclusion : MRP is an adhesin of SS2 and galactose can block its adhesion. Key words : Streptococcus suis type 2 ; cell adhesion ; adhesin ; muramidase released protein 2 ( Streptococcus suis type 2, SS2),, 1990 SS2 [1 ], 1998, [2 ] (MRP), , SS2, MRP [3,4 ] [5,6, SS2 ] MRP, : : 973 ( G ) : (1968 ),,, E2mail : 3 Corresponding author, Tel :

2 68 25 [5,6 PCR ELISA ], MRP SS2,, MRP, MRP SS HA9801 (MRP + [2 ) ], SH (MRP - ) [7 ] THB, h ; THB, 18 h ; 1 % THB, 12 h r min min, 011 mol L - 1 PBS (ph 714) CFU ml HEp 2, 10 % NCS RPMI 1640,, 5 % CO 2, 37, 18 h , 3 ; 24, Charland [4 ] 24, 015 ml, HA9801 SH L, 37 1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12 h, PBS 3, mg ml - 1 EDTA,, 015 ml , 100 L THB, h 114 HA9801 HA9801 4, 15, 28, 37, h, HA9801 Haataja [9 ] [10 ], CFU ml PBS : 0105 mol L - 1 ( ) ml - 1, ml - 1 ( Sigma ), 0122m ; RPMI g ml g DNase 0105 mol ml mg ml - 1 ( Bolinman ) 11512, HA9801, 37 1 h ; PBS 10 8 CFU ml - 1, HEp %, ( ) DNase ( ) ( ) ( 1,4 ), 015 ml h HA HA ; mg ml mol L - 1, , 20, 30, 40, 50, 60 min MRP : MRPHA9801 (MRP + ) SH (MRP - ) (, ), 1 4, , - 20 MRP : HA9801 ba1b/ c ml - 1

3 4 : 2 69 (, ), SP2/ 0, MRP, ELISA 3 3A3 4D5 ; ba1b/ c,, MRP HA9801, g MRP, ph 3pH 10 ph 3pH 5 ( The Rotofor System BIO RAD) MRP (BIO RAD/ quantity one ) SDS PAGE, MRP 014 mg ml - 1, MRP 0122m, 011 mol L - 1 PBS (ph 714) 40g ml :, HA9801 SH006444, 37 1 h 24, MRP : 24, 015 ml, MRP 50 L,, 37 1 h, HA9801 SH006444, 6, 118 MRP MRP, 5, 7 ; 56, 15 ; mg ml - 1 (MRP 1 1) ;, 37 0, 10, 20, 30, 40, min, MRP HA9801 (MRP + ) SH (MRP - ) (1), HEp 2, MRP + MRP - ( P < 0105) MRP SS2,, 6 h, MRP - 10 h 212 HA , 45 ( P < 0101), 56 (2) 1 SS2 HEp Fig. 1 Adhesive kinetics of two SS2 strains to HEp 2 2 HA cells 213 Fig. 2 Effects of pretreatment temperature on adhesion of HA9801 to HEp HA9801, 2 cells

70 25 DNase, (3) ;, ;, 1, 4 214 HA9801,,, 1 h 10 min,, 50 min, 20 min, 50 min (4) 3 HA9801 4 Fig. 3 Effects of chemicals on adhesion of HA9801 to HEp22 cells 215 MRP Fig.

4 70 25 DNase, (3) ;, ;, 1, HA9801,,, 1 h 10 min,, 50 min, 20 min, 50 min (4) 3 HA Fig. 3 Effects of chemicals on adhesion of HA9801 to HEp22 cells 215 MRP Fig. 4 Sensitivity of adhesive ability of HA9801 to pretreatmemt of trypsin, galactose and heat HA9801 ;MRP 3A3 4D5 MRP - ( MRP/ MRP -,MRP/ 3A3,MRP/ 4D5), HA9801 SH ; MRP ( MRP - Pr) MRP MRP MRP + ( P < 0105), MRP - MRP MRP +,, ;MRP - MRP -, MRP +, SS2 MRP 5 MRP HA9801, MRP Fig. 5 Inhibition of MRP and antibodies against adhesion (5) of HA9801 to HEp 2 cells 216 MRP minMRP, HA9801,

5 4 : 2 71 ( P < 0105), MRP HEp 2 56 MRP, HA9801,, 40 min, HA9801, 40 min MRP (6) 3, SS2 [9 ] MRP MRP + MRP - 6 MRP Fig. 6 Effects of heat and chemicals on the inhibitive, MRP MRP, SS2, activities of MRP against MRP, SS [11,12 ] HA9801 MRP [11 Haataja, ] ; Kurl [13 ], B CG SS2 ; SS2 N, MRP, HA9801 MRP, MRP MRP, : [1 ],,,. 2 [J ]., 1993(5) : 13. [2 ],, [J ]., 1999, 22 (2) : [3 ],,, 1 2 (MRP) PCR [J ]., 2001, 23(1) : [4 ] Charland N, Nizet V, Rubens C E, et al. Streptococcus suis serotype 2 interactions with human brain microvasc Lar endothelial cells [J ]. Immun, 2000, 68(2) : [5 ] Wisselink H J, Smith H E, Stockhofe2zurwieden N, et al. Distribution of capsular types and production of muramidase2released protein (MRP) and extracellular factor ( EF) of Streptococcus suis strains isolated from diseased pigs in seven European countries [J ]. Vet Microbiol, 2000, 74 (3) : [6 ] Berthelot2Hera Lt F, Morvan H, Keribin A M, et al. Production of muraminidase2released protein (MRP), extracellular factor ( EF) and suilysin by field isolates of Streptococcus suis capsular types 2, 1/ 2, 9, 7 and 3. isolated from wine in France [J ]. Vet Res, 2000, 31 (5) : [7 ], 1 2 ELISA [J ]., 2001, 24 (2) : [8 ],,, 1 [J ]., 2001, 21 (1) : [9 ] Haataja S, Tikkanen K, Liukkonen J, et al. Characterization of a novel bacterial adhesion specificity of Streptococcus suis recognizing blood group P receptor oligosaccharides [J ]. J Biol Chem, 1993, 268 : [10 ],, 1 MF1 [J ]., 1999, 22 (3) : [11 ] Lalonde M, Segura M, Lacouture S, et al. Interactions between Streptococcus suis serotype 2 and different epithelial cell lines [J ]. Microbiology, 2000, 146 (8) : [12 ] Quessy S, Busque P, Higgins R, et al. Description of an albumin binding activity for Streptococcus suis serotype 2 [J ]. FEMS Microbiol Lett, 1997, 147 (2) : [ 13 ] Kurl D N, Haataja S, Finne J. Hemagglutination activities of group B, C, D, and G streptococci : demonstration of novel sugar2specific cell2bind2 ing activities in Streptococcus suis [J ]. Infect Immun, 1989, 57 (2) : Infect :

Versatile antibody-sensing Boolean logic for simultaneous. detection of multiple bacterial toxins

This journal is The Royal Society of Chemistry 0 Supporting information------ Versatile antibody-sensing Boolean logic for simultaneous detection of multiple bacterial toxins Kui Zhu, Richard Dietrich,