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J B Kaper

Publications and source records attributed to J B Kaper.

At least 163 records · Page 9Linked to original sources
PubMed1989

Conserved sequences in bacterial and viral sialidases.

The genes of the bacterial sialidases from Clostridium sordellii G12, C. perfringens A99, Salmonella typhimurium LT-2 and Vibrio cholerae 395 sequenced so far were examined for homologies and were compared with sequences of viral sialidases. Each of the bacterial sialidases contains a short sequence of twelve amino-acids, which is repeated at four positions in the protein. All these sequences exhibit significant similarities. Comparing the repeated sequences of the four sialidases, five amino-acids were found to be highly conserved at defined positions: Ser-X-Asp-X-Gly-X-Thr-Trp. Additionally, most of the distances between the four repeated regions are also conserved among the different sialidases. The conserved bacterial sequences show similarity with sialidases of influenza A H7N1 and H13N9.

Amino Acid Sequence↗
PubMed1989

Enteroaggregative Escherichia coli associated with persistent diarrhea in a cohort of rural children in India.

A cohort of 452 rural children was followed longitudinally for 13 mo to ascertain the role of HEp-2 cell adherent Escherichia coli and other pathogens in causing acute (less than or equal to 14 d) and persistent (greater than 14 d) diarrhea. Aeromonas, Campylobacter jejuni, E. coli manifesting localized adherence to HEp-2 cells and enterotoxigenic E. coli were significantly associated with acute diarrhea. E. coli strains that exhibit aggregative adherence, so-called enteroaggregative E. coli, a newly-described category of diarrheagenic E. coli distinct from enterotoxigenic, enteroinvasive, enterohemorrhagic, and enteropathogenic E. coli, were found significantly more often in patients with persistent diarrhea (29.5%) than with acute diarrhea (12.8%) (P = .0052) or controls (9.9%) (P = .0006).

Acute Disease↗
PubMed1989

Human immune response to Vibrio cholerae O1 whole cells and isolated outer membrane antigens.

The serum immunoglobulin G (IgG) and mucosal secretory IgA (SIgA) response of human volunteers challenged with Vibrio cholerae O1 was analyzed for reactivity to V. cholerae O1 antigens by the immunoblot technique. Components of both in vitro- and in vivo (rabbit ligated ileal loop)-grown V. cholerae O1 were separated by sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis. Postchallenge serum IgG reacted uniquely with 15 antigens and with greater intensity than did prechallenge serum with at least 16 antigens. Serum IgG and SIgA reacted with antigens present in preparations from the homologous challenge strain of V. cholerae as well as antigens from strains of heterologous biotype or serotype. These heterologous antigens may represent antigens responsible for protection to rechallenge with a heterologous strain of V. cholerae. All the antigens detected by postchallenge jejunal fluid SIgA had an apparent molecular size of less than 25 kilodaltons. Serum IgG and jejunal fluid SIgA also reacted with antigens unique to in vivo-grown cells and several antigens in outer membrane preparations, suggesting that studies of protective immunity and V. cholerae O1 pathogenesis should include examination of both in vitro- and in vivo-grown V. cholerae O1 cellular antigens.

Animals↗
PubMed1989

Development and testing of a synthetic oligonucleotide probe for the detection of pathogenic Yersinia strains.

A 24-base oligonucleotide probe specific for a region of the Yersinia enterocolitica virulence plasmid (pYV) associated with HEp-2 cell cytotoxicity and the Sereny reaction was constructed by using sequences flanking critical TnphoA insertions in a subcloned fragment of pYV. This probe, highly specific and sensitive for virulent yersiniae, detected pathogenic Y. enterocolitica isolates in artificially inoculated foods.

Base Sequence↗
PubMed1989

Descriptive epidemiology of persistent diarrhoea among young children in rural northern India.

In order to determine the descriptive epidemiology of persistent diarrhoea in rural northern India, a cohort of 963 children aged 0-71 months was followed prospectively for 12 months through weekly household visits. The incidence of persistent diarrhoea was 6.3 per 100 child-years among those aged 0-71 months, and was highest (31 per 100 child-years) among those aged 0-11 months. There were no significant sex-related differences in the incidence of the disease, and the overall seasonal distribution of acute and persistent diarrhoea was similar. The persistence of diarrhoeal symptoms was significantly correlated with a higher initial mean stool frequency (P less than 0.01) and passage of gross blood with stools (P less than 0.001). Persistent diarrhoea was an important problem among children during the first 2 years of life. Established enteric pathogens were isolated during the initial illness in 46.4% of persistent and 55.4% of acute episodes. Pathogens isolated during persistent episodes included enterotoxigenic Escherichia coli (ETEC 9.3%), Salmonella spp. (4.7%), as well as campylobacter (4.7%), Shigella spp. (2.3%), Entamoeba histolytica (2.3%), and rotavirus (2.3%). Similar proportions of these pathogens were isolated also during episodes of acute diarrhoea. Multiple pathogens were isolated in 7% of the persistent and 5% of the acute episodes. E. coli that manifested aggregative adherence (EAEC-A) was more common (34.9% versus 12.3%) in persistent than acute episodes (P less than 0.01), and initial faecal excretion of EAEC-A was significantly associated with the persistence of a diarrhoeal episode.

Child↗
PubMed1988

Safety, immunogenicity, and efficacy of recombinant live oral cholera vaccines, CVD 103 and CVD 103-HgR.

The genes encoding the A (toxic) subunit of cholera toxin were deleted from pathogenic Vibrio cholerae O1 strain 569B by recombinant techniques, leaving intact production of immunogenic, non-toxic B subunit. The resultant strain, CVD 103, evaluated for safety, immunogenicity, and efficacy as a live oral vaccine, was highly attenuated and elicited strong antibacterial and antitoxic immune responses; a single dose significantly protected volunteers against challenge with pathogenic V cholerae O1 of either serotype or biotype. A further derivative, CVD 103-HgR, which has an Hg++-resistance gene to differentiate it from wild-type vibrios, was also well-tolerated, immunogenic, and protective; moreover, faecal excretion of this derivative was significantly lower than that of CVD 103, which should minimise environmental spread of the vaccine. CVD 103-HgR is a candidate for expanded clinical trials in endemic areas.

Administration, Oral↗
PubMed1988

Morphological studies on fimbriae expressed by Vibrio cholerae 01.

Colonization of the small intestine is an essential step in the pathogenesis of Vibrio cholerae diarrhea. At least one type of fimbriae, known as TcpA are required for the colonization process. This paper reports electron microscopic evidence that V. cholerae strains can express at least two other fimbrial types. Classical strains express three types: TcpA fimbriae are 5-6 nm in diameter and form bundles of parallel undulating filaments up to 15 micron long; Type B are 3 nm wide and of wavy morphology, and Type C are rigid, isolated filaments 5-6 nm wide and 180-800 nm long. El Tor strains express fimbriae resembling Types B and C. Types B and C were also found on a tcpA- isogenic mutant of V. cholerae 395 N1, and are thus encoded by genetically distinct loci. TcpA fimbriae, but not Types B or C, were labeled with gold-conjugated anti-TcpA antibody. Four El Tor strains, including two environmental isolates that poorly colonize humans, expressed fimbriae resembling Types B and C, but did not express TcpA. Multiple types of fimbriae may represent colonization factors for surfaces present in the environment and in the human gut. Characterization of the role of fimbriae in immunogen presentation and immunity could facilitate the improvement of cholera vaccines.

Cholera↗
PubMed1988

Progress towards a vaccine against enterotoxigenic Escherichia coli.

A variety of approaches are being investigated in the development of a vaccine against enterotoxigenic Escherichia coli (ETEC). These approaches include purified fimbriae vaccines, toxoid vaccines, live attenuated E. coli vaccine strains and ETEC antigens expressed in carrier organisms. Studies of the pathogenesis and immune response to ETEC indicate that development of a vaccine against human ETEC is a realistic goal but considerable work remains before this goal is realized.

Bacterial Vaccines↗
PubMed1988

Characterization of enteroadherent-aggregative Escherichia coli, a putative agent of diarrheal disease.

Escherichia coli that exhibit the aggregative pattern of adherence to HEp-2 cells (enteroadherent-aggregative E. coli [EA-AggEC]) have been epidemiologically incriminated as a cause of diarrhea. We undertook a preliminary microbiological and pathogenetic characterization of 42 isolates of this putative pathogen. The strains were negative by tests with DNA probes for enteropathogenic, enterotoxigenic, enteroinvasive, and enterohemorrhagic E. coli and, by serotype, did not fit these categories. Thirty-nine of 42 strains had a 55-65-megadalton plasmid; many shared DNA homology. With one representative strain, plasmid transfer was accompanied by transfer of smooth lipopolysaccharide, fimbriae expression, and the aggregative property. EA-AggEC caused characteristic lesions in rabbit and rat ileal loops. The intestinal lesions and (Shiga-like) limb paralysis and death in rabbits inoculated with live organisms suggest toxin involvement; assays for Shiga-like toxins were negative. These preliminary results support the contention that EA-AggEC may represent a distinct category of diarrheagenic E. coli.

Animals↗
PubMed1988

Toxin profiles of Vibrio cholerae non-O1 from environmental sources in Calcutta, India.

A collection of Vibrio cholerae non-O1 isolated from the aquatic environs of Calcutta, a cholera-hyperendemic area, were examined for the production of cholera toxin (CT), Shiga-like toxins (Vero toxins), heat-stable enterotoxin, and hemolysins. Two (0.5%) V. cholerae non-O1 isolates produced CT. The DNA from both these isolates also hybridized with a DNA probe containing sequences encoding the A subunit of CT. None of the strains produced Shiga-like toxins or heat-stable enterotoxin. Hemolytic activity was observed in 89.7% of the strains, of which 36.1% exhibited biological activity in the suckling mouse. However, none of them produced a hemolysin that cross-reacted with the thermostable direct hemolysin of Vibrio parahaemolyticus. It appears from this study that a small percentage of environmental V. cholerae non-O1 strains do possess the potential for causing cholera-like diarrhea.

Cholera↗
PubMed1988

Determinants of immunogenicity and mechanisms of protection by virulent and mutant Vibrio cholerae O1 in rabbits.

The colonizing capacities of 16 Vibrio cholerae strains, including nine genetically and/or phenotypically defined parent-mutant pairs, were determined in unobstructed adult rabbit small bowel. There were marked interstrain differences in colonizing capacity, which was enhanced by bacterial motility and the production of cholera holotoxin but was unrelated to production of cholera toxin B-subunit or hemolysin or to bacterial serotype or biotype. The role of colonizing capacity and other bacterial features in determining the immunizing efficiency of live V. cholerae was studied by determining the efficiency with which graded inocula of each strain immunized against attempted recolonization of the ileum or induction of choleralike diarrhea by the RITARD (removable intestinal tie-adult rabbit diarrhea) challenge technique using standard inocula of virulent V. cholerae. Mucosal colonizing capacity was the only quantitative predictor of bacterial immunizing capacity; none of the other bacterial features cited above influenced bacterial immunogenicity against either type of challenge, except as they affected colonizing capacity. Live V. cholerae immunized much more efficiently than Formalin-killed bacteria; the former caused marked protection after a single inoculum of 10(2) CFU, whereas the latter gave only partial protection after three inoculations of 10(11) killed organisms. Protection induced by live bacteria was due largely to resistance to colonization and included marked inhibition of bacterial growth within the bowel lumen. These findings strongly suggest that an optimally efficient oral cholera vaccine would be composed of avirulent live V. cholerae selected for their capacity to colonize the small-bowel mucosa.

Administration, Oral↗
PubMed1988

Volunteer studies of deletion mutants of Vibrio cholerae O1 prepared by recombinant techniques.

Vibrio cholerae O1 A-B- vaccine strain JBK 70 and A-B+ CVD 101 prepared by recombinant DNA techniques from pathogenic EI Tor Inaba N16961 and classical Ogawa 395, respectively, were fed to 38 volunteers in single doses of 10(4) to 10(10). Although severe diarrhea did not occur in any vaccine, more than one-half developed mild diarrhea. These attenuated strains colonized well and elicited prominent vibriocidal and antitoxic (CVD 101) antibody responses. Recipients of a single dose of JBK 70 were significantly protected when challenged with 10(6) wild-type N16961. Diarrhea occurred in 7 of 8 controls but in only 1 of 10 vaccines (P less than 0.003, 89% vaccine efficacy), demonstrating the potency of immune mechanisms that do not involve cholera antitoxin. Further derivatives were prepared to explore the pathogenesis of the residual diarrhea, considering that either intestinal colonization by the vaccine itself or accessory toxins might be responsible. CVD 102, an auxotrophic mutant of CVD 101, did not cause diarrhea but colonized poorly and elicited feeble immune responses. Derivatives of JBK 70 and CVD 101 (CVD 104 and 105) deleted of genes encoding the EI Tor hemolysin still caused mild diarrhea. Genetically engineered strains can be colonizing, highly immunogenic, and protective single-dose oral vaccines, but they must be further attenuated before they can be considered for use as public health tools.

ABO Blood-Group System↗
PubMed1988

Cloning and expression of the Vibrio cholerae neuraminidase gene nanH in Escherichia coli.

A cosmid gene bank of Vibrio cholerae 395, classical Ogawa, was screened in Escherichia coli HB101 for expression of the vibrio neuraminidase (NANase) gene nanH (N-acylneuraminate glycohydrolase). Positive clones were identified by their ability to cleave the fluorogenic NANase substrate 2'-(4-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid. Seven NANase-positive clones were detected after screening 683 cosmid isolates with a rapid, qualitative plate assay method. The nanH gene was subcloned from one of the cosmids and was located within a 4.8-kilobase-pair BglII restriction endonuclease fragment. Evidence that nanH was the NANase structural gene was obtained by transposon mutagenesis and by purification and comparison of the cloned gene product with the secreted NANase purified from the parent V. cholerae strain. The sequence of the first 20 amino-terminal amino acids of the secreted NANase purified from V. cholerae was determined by automated Edman degradation and matched perfectly with the amino acid sequence predicted from nucleotide sequencing of nanH. The sequence data also revealed the existence of a potential signal peptide that was apparently processed from NANase in both V. cholerae and E. coli. In contrast to V. cholerae, E. coli nanH+ clones did not secrete NANase into the growth medium, retaining most of the enzyme in the periplasmic compartment. Kinetic studies in V. cholerae showed that nanH expression and NANase secretion were temporally correlated as cells in batch culture entered late-exponential-phase growth. Similar kinetics were observed in at least one of the E. coli nanH+ clones, suggesting that nanH expression in E. coli might be controlled by some of the same signals as in the parent V. cholerae strain.

Amino Acid Sequence↗
PubMed1988

The current status of cholera vaccine development and experience with cholera vaccine trials in volunteers.

In recent years notable advances have been made in the development of improved vaccines to prevent cholera. These new vaccines are administered orally to maximally stimulate intestinal secretory immunity. Killed vibrios, given in conjunction with purified B subunit or administered alone, in three spaced doses, caused no adverse reactions and have conferred significant protection in volunteer challenge studies and in field trials. Two attenuated mutants of V. cholerae, prepared by recombinant DNA techniques, CVD 103 and CVD 103-HgR are well-tolerated and elicit prominent immune responses and protective immunity after ingestion of a single oral dose. Other modern approaches being pursued include the development of auxotrophic strains and of modifying attenuated S. typhi strain Ty21a to express V. cholerae Inaba and Ogawa LPS antigens.

Administration, Oral↗
PubMed1987

Serological response to type 1-like somatic fimbriae in diarrheal infection due to classical enteropathogenic Escherichia coli.

Fimbriae from enteropathogenic Escherichia coli strain E2349/69 (0127:H6) and its plasmid-minus derivative, MAR20, were purified and characterized as type 1-like by their physicochemical and hemagglutination patterns. Sera from adult volunteers challenged with the diarrheagenic parent strain and the attenuated plasmid-minus derivative were examined to detect an immune response, using the purified fimbriae as antigens in an enzyme linked immunosorbent assay (ELISA) and immunoblot assay. An anti-fimbrial response was evident in sera of 7 of 10 volunteers fed the diarrheagenic parent strain E2348 but also in 8 of 9 individuals fed the attenuated, plasmid-cured, derivative MAR20. The antibody response appeared specific in that the sera failed to react in an ELISA and by immunoblot assay with type 1 fimbriae from other E. coli. These findings suggest that the type 1 fimbriae of this representative EPEC strain are antigenically distinct. The results of this investigation provide the first evidence of seroconversion to type 1-like fimbriae in infections caused by diarrheagenic E. coli.

Adult↗
PubMed1987

A DNA probe to identify enterohemorrhagic Escherichia coli of O157:H7 and other serotypes that cause hemorrhagic colitis and hemolytic uremic syndrome.

Enterohemorrhagic Escherichia coli (EHEC) cause hemorrhagic colitis and hemolytic uremic syndrome (HUS), make potent cytotoxins (Verotoxins [VT] or Shiga-like toxins), and possess a plasmid (approximately 60 megadaltons) that encodes a new fimbrial antigen and promotes attachment to epithelial cells. We evaluated the use of a DNA probe, prepared from a 3.4-kilobase segment of the EHEC plasmid, to detect EHEC. The probe hybridized with 106 (99%) of 107 O157:H7 and 34 (77%) of 44 O26:H11, VT-positive strains from patients with colitis, HUS, and diarrheal disease and hybridized with 21 (81%) of 26 VT-positive E. coli of serotypes other than O157:H7 or O26:H11 from patients with hemorrhagic colitis and HUS. We examined 601 other strains, including 18 serotype O26 isolates of H types other than H11, 306 enteropathogenic E. coli, 60 enteroinvasive E. coli, 119 enterotoxigenic E. coli, and 20 isolates from the urinary tract and 77 isolates from the normal intestinal flora; only one (O127:H-) was positive (specificity, 99.8%). Serotype O26:H11, previously considered a classic enteropathogenic E. coli serotype, is now shown to be EHEC.

Child↗
PubMed1987

Patterns of adherence of diarrheagenic Escherichia coli to HEp-2 cells.

A total of 516 Escherichia coli strains randomly isolated from coprocultures of 154 Chilean children with diarrhea and 66 controls were examined with DNA probes and tested for adherence to HEp-2 cells. Three adherence patterns were distinguished, localized, true diffuse and "aggregative." Enteropathogenic E. coli (EPEC) were detected by EPEC adherence factor probe among 86 of the 372 isolates (23%) from patients with diarrhea vs. 14 of 144 (10%) strains from controls (P less than 0.0002). Of 95 strains that manifested localized adherence, 97% were EPEC adherence factor probe-positive; thus the HEp-2 assay may serve as an alternative to the probe in identifying EPEC adherence factor-positive EPEC. True diffuse adherence was not associated with diarrhea. In contrast the aggregative pattern appears to signify a new, distinct class of diarrheagenic E. coli (enteroadherent-aggregative E. coli). The aggregative pattern was found in only 3 of 27 enterotoxigenic, 0 of 4 enteroinvasive, 0 of 2 enterohemorrhagic and 2 of 86 EPEC strains but in 84 of 253 probe-negative strains (P less than 0.00001) from patients with diarrhea; in comparison only 20 of 134 probe-negative strains from controls were aggregative E. coli (P less than 0.00001 vs. probe-negative strains from diarrhea patients).

Bacterial Adhesion↗