Confocal microscopy study of pertussis toxin and toxoids on CHO-cells

被引：5

作者：

Tan, Yajun ^{[1
,2
]}

Fleck, Roland A. ^{[3
]}

Asokanathan, Catpagavalli ^{[3
]}

Yuen, Chun-Ting ^{[3
]}

Xing, Dorothy ^{[3
]}

Zhang, Shumin ^{[1
,2
]}

Wang, Junzhi ^{[1
]}

机构：

[1] Natl Inst Food & Drug Control, Beijing, Peoples R China

[2] Peking Union Med Coll, Grad Sch, Beijing 100021, Peoples R China

[3] Natl Inst Biol Stand & Controls, Potters Bar, Herts, England

来源：

HUMAN VACCINES & IMMUNOTHERAPEUTICS | 2013年 / 9卷 / 02期

关键词：

confocal microscopy; pertussis toxin; toxoid; detoxification; CHO cells; translocation; ISLET-ACTIVATING PROTEIN; HISTAMINE-SENSITIZATION TEST; ADP-RIBOSYLATION ACTIVITY; VITRO ASSAY SYSTEM; B-OLIGOMER; HYDROGEN-PEROXIDE; WHOOPING-COUGH; VACCINE; FORMALDEHYDE; BINDING;

D O I：

10.4161/hv.22795

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Pertussis toxin in its detoxified form is a major component of all current acellular pertussis vaccines. Here we report the membrane translocation and internalization activities of pertussis toxin and various pertussis toxoids using Chinese hamster ovary cells and confocal microscopy based on indirect immunofluorescence labeling. Chemically detoxified pertussis toxoids were able to translocate/internalize into cells at the concentration about 1,000 times higher than the native toxin. Pertussis toxoids detoxified with different procedures (glutaraldehyde, glutaraldehyde plus formaldehyde, hydrogen peroxide or genetic mutation) showed differences in fluorescence intensity under the same condition, indicating toxoids from different detoxification methods may have different translocation/ internalization activities on cells.

引用

页码：332 / 338

页数：7

共 36 条

[1] [Anonymous], 2000, EUR PHARMACOPOEIA S, P1050
[2] [Anonymous], 1998, WHO TECHNICAL REPORT, V878, P57
[3] [Anonymous], 2005, WEEKLY EPIDEMIOLOGIC
[4] [Anonymous], 2004, MIN REQ BIOL PROD, P41
[5] A quantitative analysis for the ADP-ribosylation activity of pertussis toxin: An enzymatic-HPLC coupled assay applicable to formulated whole cell and acellular pertussis vaccine products
Cyr, T
Menzies, AJ
Calver, J
Whitehouse, LW
[J]. BIOLOGICALS, 2001, 29 (02) : 81 - 95
[6] EDWARDS KM, 1995, PEDIATRICS, V96, P548
[7] ADP-RIBOSYLATION OF ALPHA(I3)C20 BY THE S1 SUBUNIT AND DELETION PEPTIDES OF S1 OF PERTUSSIS TOXIN
FINCKBARBANCON, V
BARBIERI, JT
[J]. BIOCHEMISTRY, 1995, 34 (03) : 1070 - 1075
[8] Novel configurations of high molecular weight species of the pertussis toxin vaccine component
Fowler, S
Byron, O
Jumel, K
Xing, D
Corbel, MJ
Bolgiano, B
[J]. VACCINE, 2003, 21 (19-20) : 2678 - 2688
[9] Modifications of the catalytic and binding subunits of pertussis toxin by formaldehyde: effects on toxicity and immunogenicity
Fowler, S
Xing, DKL
Bolgiano, B
Yuen, CT
Corbel, MJ
[J]. VACCINE, 2003, 21 (19-20) : 2329 - 2337
[10] ADP-ribosylation activity in pertussis vaccines and its relationship to the in vivo histamine-sensitisation test
Gomez, S. R.
Yuen, C.-T.
Asokanathan, C.
Douglas-Bardsley, A.
Corbel, M. J.
Coote, J. G.
Parton, R.
Xing, D. K. L.
[J]. VACCINE, 2007, 25 (17) : 3311 - 3318

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