Transcription profile of Helicobacter pylori in the human stomach reflects its physiology in vivo

被引:23
作者
Booniakuakul, JK
Syvanen, M
Suryaprasad, A
Bowlus, CL
Solnick, JV [1 ]
机构
[1] Univ Calif Davis, Ctr Comparat Med, Davis, CA 95616 USA
[2] Univ Calif Davis, Dept Med Microbiol & Immunol, Davis, CA 95616 USA
[3] Univ Calif Davis, Dept Internal Med, Davis, CA 95616 USA
关键词
D O I
10.1086/423142
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Background. Little is known about levels of expression of Helicobacter pylori genes in the human host. We therefore developed a quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) assay to measure transcript profiles of H. pylori in the human stomach. Methods. In vivo expression of 16 genes on the cag pathogenicity island and of 18 putative virulence genes was quantitated by isolation of total RNA directly from infected human gastric mucosa. The results were compared with in vitro expression determined from H. pylori cells grown in culture. Results. The highest levels of expression were found for cag1 and cag25 and for genes, such as urease and catalase, that may be important for bacterial homeostasis in the relatively hostile environment of the gastric mucosa. Transcript abundance, relative to 16S rRNA, was lower in vivo than in vitro, which suggests that H. pylori cells are in stationary phase in the gastric environment. This was particularly apparent for cagA. Since CagA is arguably of unique importance, in terms of interaction with the host, tight control of its in vivo expression might be particularly important. Conclusions. qRT-PCR is a powerful tool to measure gene expression in human or animal tissue that contains minute amounts of microbial mRNA, and the results reflect on the physiology of the pathogen in its natural host.
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页码:946 / 956
页数:11
相关论文
共 56 条
[1]   Genomic-sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori [J].
Alm, RA ;
Ling, LSL ;
Moir, DT ;
King, BL ;
Brown, ED ;
Doig, PC ;
Smith, DR ;
Noonan, B ;
Guild, BC ;
deJonge, BL ;
Carmel, G ;
Tummino, PJ ;
Caruso, A ;
Uria-Nickelsen, M ;
Mills, DM ;
Ives, C ;
Gibson, R ;
Merberg, D ;
Mills, SD ;
Jiang, Q ;
Taylor, DE ;
Vovis, GF ;
Trost, TJ .
NATURE, 1999, 397 (6715) :176-180
[2]   MOSAICISM IN VACUOLATING CYTOTOXIN ALLELES OF HELICOBACTER-PYLORI - ASSOCIATION OF SPECIFIC VACA TYPES WITH CYTOTOXIN PRODUCTION AND PEPTIC-ULCERATION [J].
ATHERTON, JC ;
CAO, P ;
PEEK, RM ;
TUMMURU, MKR ;
BLASER, MJ ;
COVER, TL .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (30) :17771-17777
[3]   Density of Helicobacter pylori infection in vivo as assessed by quantitative culture and histology [J].
Atherton, JC ;
Tham, KT ;
Peek, RM ;
Cover, TL ;
Blaser, MJ .
JOURNAL OF INFECTIOUS DISEASES, 1996, 174 (03) :552-556
[4]  
Ausubel F., 1990, CURRENT PROTOCOLS MO
[5]   Dynamics of Helicobacter pylori colonization in relation to the host response [J].
Blaser, MJ ;
Kirschner, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (15) :8359-8364
[6]   Ecology of Helicobacter pylori in the human stomach [J].
Blaser, MJ .
JOURNAL OF CLINICAL INVESTIGATION, 1997, 100 (04) :759-762
[7]   Modelling carbon responses of tundra ecosystems to historical and projected climate: sensitivity of pan-Arctic carbon storage to temporal and spatial variation in climate [J].
McGuire, AD ;
Clein, JS ;
Melillo, JM ;
Kicklighter, DW ;
Meier, RA ;
Vorosmarty, CJ ;
Serreze, MC .
GLOBAL CHANGE BIOLOGY, 2000, 6 :141-159
[8]   cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and disease-associated virulence factors [J].
Censini, S ;
Lange, C ;
Xiang, ZY ;
Crabtree, JE ;
Ghiara, P ;
Borodovsky, M ;
Rappuoli, R ;
Covacci, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (25) :14648-14653
[9]   NapA protects Helicobacter pylori from oxidative stress damage, and its production is influenced by the ferric uptake regulator [J].
Cooksley, C ;
Jenks, PJ ;
Green, A ;
Cockayne, A ;
Logan, RPH ;
Hardie, KR .
JOURNAL OF MEDICAL MICROBIOLOGY, 2003, 52 (06) :461-469
[10]   Did the inheritance of a pathogenicity island modify the virulence of Helicobacter pylori? [J].
Covacci, A ;
Falkow, S ;
Berg, DE ;
Rappuoli, R .
TRENDS IN MICROBIOLOGY, 1997, 5 (05) :205-208