Abstract:
:Mycobacterium tuberculosis, the primary etiologic agent of tuberculosis, is the world's leading cause of death from a single infectious agent, and new vaccines and drugs to combat it are urgently needed. The major extracellular proteins of M. tuberculosis, which are released into its phagosome in macrophages, its host cells in humans, are leading candidates for a vaccine and prime targets for new drugs. However, the development of these biologicals has been hampered by the unavailability of large quantities of recombinant extracellular proteins identical to their native counterparts. In this report, we describe the heterologous expression and secretion of four major M. tuberculosis extracellular proteins (the 30-, 32, 16-, and 23.5-kDa proteins--the first, second, third, and eighth most abundant, respectively) in rapidly growing, nonpathogenic mycobacterial species. Multiple attempts to obtain secretion of the proteins by using Escherichia coli- and Bacillus subtilis-based expression systems were unsuccessful, suggesting that high-level expression and secretion of these Mycobacterium-specific proteins require a mycobacterial host. All four recombinant proteins were stably expressed from the cloned genes' own promoters at yields that were 5- to 10-fold higher than those observed for the native proteins. The four proteins were purified to apparent homogeneity from culture filtrates by ammonium sulfate precipitation and ion-exchange and molecular sieve chromatography. The recombinant proteins were indistinguishable from their native counterparts by multiple criteria. First, N-terminal amino acid sequence determination demonstrated that processing of the leader peptides was highly accurate. Second, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed identical migration patterns. Third, mass spectrometry analysis confirmed that differences in mass were < or = 5 Da. A homolog of the M. tuberculosis 30-kDa protein was identified in M. smegmatis by means of DNA analyses and immunoscreening. This is the first time that secretion of recombinant M. tuberculosis extracellular proteins in their native form has been achieved. This study opens the door to mass production of correctly processed and secreted extracellular proteins of M. tuberculosis in a heterologous host and allows ready evaluation of their biologic and immunologic function.
journal_name
Infect Immunjournal_title
Infection and immunityauthors
Harth G,Lee BY,Horwitz MAdoi
10.1128/IAI.65.6.2321-2328.1997subject
Has Abstractpub_date
1997-06-01 00:00:00pages
2321-8issue
6eissn
0019-9567issn
1098-5522journal_volume
65pub_type
杂志文章abstract::Patent filarial infection is associated with the downregulation of parasite-specific immune reactivity. In the present study, the relationship between in vitro parasite antigen-specific humoral and cellular immune responsiveness was investigated in Brugia pahangi-infected jirds and in jirds immunized with soluble anti...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.56.12.3052-3057.1988
更新日期:1988-12-01 00:00:00
abstract::A genomic library of meningococcal DNA from a clinical isolate of Neisseria meningitidis was constructed in the expression vector lambda gt11. Outer membrane complex was prepared from the same strain and used to immunize rabbits to raise polyclonal anti-outer membrane complex serum. The amplified library was probed wi...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.55.11.2734-2740.1987
更新日期:1987-11-01 00:00:00
abstract::Three-month-old male Crl:CD1(ICR)BR and C3H/HeJ mice were fed chow containing Candida albicans for 14 days, while similar control mice were fed regular food. Stool cultures were done for all mice before and after administration of the special diet. Stool cultures were repeated 48 h, 1 week, and 1 and 3 months after st...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.58.6.1514-1517.1990
更新日期:1990-06-01 00:00:00
abstract::Staphylococcus aureus fibronectin-binding proteins (FnBPs) play a critical role in S. aureus pathogenesis. FnBPs mediate adhesion to fibronectin and invasion of mammalian cells, including epithelial, endothelial, and fibroblastic cells, by fibronectin bridging to the host cell fibronectin receptor integrin (alpha(5))b...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.72.12.7155-7163.2004
更新日期:2004-12-01 00:00:00
abstract::The 260-kDa galactose-inhibitable lectin of Entamoeba histolytica mediates binding of amebic trophozoites to purified colonic mucins and intestinal epithelial cells. Parenteral immunization of Lewis rats with immuno-affinity-purified lectin with Freund's adjuvant and then intra-Peyer's patch inoculation of lectin with...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.63.2.686-689.1995
更新日期:1995-02-01 00:00:00
abstract::Trefoil family factor 2 (TFF2), also known as spasmolytic peptide, is a low-molecular-weight protein that is upregulated in gastric tissues infected with Helicobacter or having other inflammatory conditions, but a precise function is yet to be elucidated. The role of TFF2 in the development of gastritis, colitis, and ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.02039-05
更新日期:2007-01-01 00:00:00
abstract::The recombinant protective antigen (rPA) of Bacillus anthracis is a promising anthrax vaccine. We compared serum immunoglobulin G levels and toxin-neutralizing antibody titers in rabbits following delivery of various doses of vaccine by microneedle-based intradermal (i.d.) delivery or intramuscular (i.m.) injection us...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.01210-06
更新日期:2006-12-01 00:00:00
abstract::The selC tRNA gene is a common site for the insertion of pathogenicity islands in a variety of bacterial enteric pathogens. We demonstrate here that Escherichia coli that produces Shiga toxin 2d and does not harbor the locus of enterocyte effacement (LEE) contains, instead, a novel genomic island. In one representativ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.69.11.6863-6873.2001
更新日期:2001-11-01 00:00:00
abstract::Staphylococcus aureus biofilm formation is induced in iron-restricted growth conditions in vitro. In this study, we showed that Emp and Eap play important roles in low-iron-induced biofilm formation of S. aureus Newman. Eap and Emp are secreted proteins which are non-covalently attached to the S. aureus cell surface a...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.01635-07
更新日期:2008-04-01 00:00:00
abstract::Scrub typhus is a potentially lethal infection that is caused by the obligate intracellular bacterium Orientia tsutsugamushi The roles of Toll-like receptor 2 (TLR2) and TLR4 in innate recognition of O. tsutsugamushi have not been elucidated. By overexpression of TLR2 or TLR4 in HEK293 cells, we demonstrated that TLR2...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00185-16
更新日期:2016-11-18 00:00:00
abstract::Vibrio cholerae is the causal organism of the diarrheal disease cholera. The rugose variant of V. cholerae is associated with the secretion of an exopolysaccharide. The rugose polysaccharide has been shown to confer increased resistance to a variety of agents, such as chlorine, bioacids, and oxidative and osmotic stre...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/iai.68.4.1967-1974.2000
更新日期:2000-04-01 00:00:00
abstract::Enteropathogenic Escherichia coli (EPEC) interacts with intestinal epithelial cells, activating host signaling pathways leading to cytoskeletal rearrangements and ultimately diarrhea. In this study, we demonstrate that EPEC interacts with the macrophage-like cell line J774A.1 to inhibit phagocytosis by these cells. An...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.67.2.490-495.1999
更新日期:1999-02-01 00:00:00
abstract::The typically recovered quantity of nontypeable Haemophilus influenzae (NTHi) bacteria in an ex vivo middle ear (ME) aspirate from the chinchilla model of experimental otitis media is insufficient for direct analysis of gene expression by microarray or of lipopolysaccharide glycoforms by mass spectrometry. This prompt...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00318-07
更新日期:2007-08-01 00:00:00
abstract::We found that thiols reduced the amount of cholera toxin produced by Vibrio cholerae 569B in vitro. A sulfhydryl group at least was necessary for the reduction of cholera toxin production by thiols. ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.53.3.700-701.1986
更新日期:1986-09-01 00:00:00
abstract::Accurate diagnosis and early treatment of tuberculosis (TB) and latent TB infection (LTBI) are vital to prevent and control TB. The lack of specific biomarkers hinders these efforts. This study's purpose was to screen immunological markers that discriminate Mycobacterium tuberculosis infection outcomes in a setting wh...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00759-17
更新日期:2018-03-22 00:00:00
abstract::The plasma level of the chemokine CCL3 is elevated in patients with chronic severe schistosomiasis mansoni. We have previously shown that CCL3(-/-) mice with experimental infection showed diminished pathology and worm burden compared to those of wild-type (WT) mice. To elucidate further the role of CC chemokines durin...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00502-10
更新日期:2011-04-01 00:00:00
abstract::Mosquitoes are infected with Plasmodium falciparum by taking a blood meal from a gametocyte carrier. Since a mosquito takes a volume of 1 to 2 microl, a blood meal may contain 1 x 10(4) to 3 x 10(4) leukocytes (WBC). The majority of WBC are composed of neutrophils which may phagocytose and kill developing gametes insi...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.65.9.3834-3837.1997
更新日期:1997-09-01 00:00:00
abstract::In its canonical role the reverse transcriptase telomerase recovers the telomeric repeats that are lost during DNA replication. Other locations and activities have been recently described for the telomerase protein subunit TERT in mammalian cells. In the present work, using biochemistry, molecular biology, and electro...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.02269-14
更新日期:2015-01-01 00:00:00
abstract::Harvard caries-resistant rats (CR) monoassociated with cariogenic Streptococcus faecalis (SF) for 150 days developed early high levels of specific SF serum agglutinins, moderate levels of salivary antibody, and little or no caries activity. Harvard caries-susceptible (CS) SF-monoassociated rats developed serum antibod...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.16.3.805-811.1977
更新日期:1977-06-01 00:00:00
abstract::Lysogenic conversion of Shigella flexneri type antigens was studied with the aid of wild-type and thermosensitive mutant phages. With all wild-type phages, the appearance of glycosylated antigen was accompanied by the appearance of polyprenyl phosphate glucose synthetase activity. With some of the mutant phages, the a...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.14.6.1290-1292.1976
更新日期:1976-12-01 00:00:00
abstract::Helicobacter pylori colonizes the gastric epithelium of at least 50% of the world's human population, playing a causative role in the development of chronic gastritis, peptic ulcers, and gastric adenocarcinoma. Current evidence indicates that H. pylori can invade epithelial cells in the gastric mucosa. However, relati...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.01085-06
更新日期:2006-12-01 00:00:00
abstract::Inoculation of herpes simplex virus on the forehead and/or snout of hairless mice resulted in a significantly lower mortality rate than inoculation of the skin in the lumbosacral area. Latent herpes simplex virus infections were detected in all forehead-inoculated and in 90% of snout-inoculated mice. Phosphonoacetic a...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.20.1.130-135.1978
更新日期:1978-04-01 00:00:00
abstract::The ability to bind and utilize hemin is a trait common to many human pathogens. Nevertheless, the relationship between Legionella pneumophila, the agent of Legionnaires' disease, and hemin has received little attention. Thus, we explored the capacity of a virulent, serogroup 1 strain of L. pneumophila to bind hemin a...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.64.3.842-848.1996
更新日期:1996-03-01 00:00:00
abstract::Malaria infection caused by Plasmodium parasites continues to cause enormous morbidity and mortality in areas where it is endemic, and there is no licensed vaccine capable of inducing sterile protection. Hyperimmunization with attenuated whole sporozoites can induce sterile protective immune responses targeting preery...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.01439-15
更新日期:2016-07-21 00:00:00
abstract::Strains of Bacteroides fragilis that produce a ca. 20-kDa heat-labile protein toxin (termed B. fragilis toxin [BFT]) have been associated with diarrheal disease of animals and humans. BFT alters the morphology of intestinal epithelial cells both in vitro and in vivo and stimulates secretion in ligated intestinal segme...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.65.3.1007-1013.1997
更新日期:1997-03-01 00:00:00
abstract::In the murine model of urinary tract infections (UTI), cystitis by uropathogenic Escherichia coli (UPEC) occurs through an intimate relationship with the bladder superficial umbrella cell entailing cycles of adherence, invasion, intracellular bacterial community (IBC) formation, and dispersal (fluxing) from the intrac...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.73.11.7657-7668.2005
更新日期:2005-11-01 00:00:00
abstract::The biochemical properties of polymorphonuclear neutrophils from blood and peritoneal exudates of rabbits were compared. All enzymes measured showed almost identical activities in both types of cells, except for alkaline phosphodiesterase, the activity of which was seven times higher in peritoneal neutrophils. During ...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:
更新日期:1980-08-01 00:00:00
abstract::Upon infection of a host, the pathogenic fungus Aspergillus fumigatus is attacked by the reactive oxygen species produced by phagocytic cells. Detoxification of hydrogen peroxide by catalases was proposed as a way to overcome this host response. A. fumigatus produces three active catalases; one is produced by conidia,...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/iai.71.6.3551-3562.2003
更新日期:2003-06-01 00:00:00
abstract::A Gram-negative, filamentous, rod-shaped bacillus which failed to grow in cell-free media was isolated in apparently pure culture from the bronchial scraping and washing of a laboratory rat suffering from chronic respiratory disease by inoculating embryonated chicken eggs via the allantoic route. None of the embryos d...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.47.2.472-479.1985
更新日期:1985-02-01 00:00:00
abstract::Species-specific antibody epitopes within several major immunoreactive protein orthologs of Ehrlichia species have recently been identified and molecularly characterized. In this study, dominant B-cell epitopes within the acidic (pI 5.35) ankyrin repeat-containing 200-kDa major immunoreactive protein (gp200) of Ehrlic...
journal_title:Infection and immunity
pub_type: 杂志文章
doi:10.1128/IAI.00041-07
更新日期:2007-10-01 00:00:00