Search PubMed⌕ Search

Biomedical subjects

A Hensel

Publications and source records attributed to A Hensel.

At least 73 records · Page 4Linked to original sources
PubMed1996

Bacterial ghosts: non-living candidate vaccines.

Expression of cloned PhiX174 gene E in bacteria results in lysis of bacteria. It is unique among phage lysis systems as it introduces a transmembrane tunnel structure through the cell envelope complex of Gram-negative bacteria. The resulting bacterial ghosts have intact envelope structures devoid of cytoplasmic contents. E-mediated lysis has been achieved in a variety of Gram-negative bacteria including Escherichia coli, Salmonella typhimurium, Vibrio cholerae, Klebsiella pneumoniae, and Actinobacillus pleuropneumoniae. Such ghosts, derived from human or animal pathogens, have been proposed as non-living candidate vaccines and represent an alternative to heat or chemically inactivated bacteria. In 'recombinant ghosts', foreign proteins (e.g., viral proteins) are inserted into the inner membrane via specific N-, or C-, or N- and C-terminal anchor sequences prior to lysis. Relevant advantages of (recombinant) bacterial ghosts as immunogens include: (i) inactivation procedures that denature relevant immunogenic determinants are not employed in the production of ghosts used as vaccines or as carriers of relevant antigens; (ii) the recombinant proteins are inserted into a highly immune stimulatory environment; (iii) there is no size limitation of the foreign protein moieties: multiple antigenic determinants can be presented simultaneously; (iv) bacterial ghosts can be produced inexpensively in large quantities; (v) (recombinant) ghosts are stable for long periods of time and do not require the cold chain storage system. Intraperitoneal, subcutaneous or intramuscular applications of recombinant ghosts in experimental animals induced specific humoral and cellular immune responses against bacterial and viral components. Initial aerosol vaccinations of swine with ghosts from Actinobacillus pleuropneumoniae showed that protective immunity can be established by this route of application and that the well-preserved surface structures of ghosts obtained by E-mediated lysis are able to target the mucosal immune system.

Animals↗
PubMed1996

Induction of protective immunity by aerosol or oral application of candidate vaccines in a dose-controlled pig aerosol infection model.

In order to outline basic concepts for the design of a bacterial aerosol infection model, the development of a pig model with Actinobacillus pleuropneumoniae is described. First, reproducibility of aerosol parameters should be maintained by optimizing generating and sampling conditions. Survival rates of the chosen strain must be predictable. Secondly, inhalation conditions for the recipients have to be standardized to enable the determination of deposition sites and the dose administered. Subsequently, dose-response relationship should be evaluated to find a suitable challenge dose. Furthermore, it seems necessary to establish methods to obtain local specimens for determination of the local immune responses. The present study demonstrates that after aerosol challenge pigs were completely protected after inhalation and partially protected after oral application of A. pleuropneumoniae vaccines and describes techniques to administer bacteria in a dose-dependent, viable way. Using the infection model several stages of the disease from acute pleuropneumonia to chronic infection can be induced for research purposes.

Actinobacillus Infections↗
PubMed1995

Transesterification reactions of parabens (alkyl 4-hydroxybenzoates) with polyols in aqueous solution.

Accelerated stability tests of aqueous solutions containing parabens and polyols were performed using concentrations similar to pharmaceutical and cosmetic formulations. Reaction products were detected in these solutions by HPLC and identified by chromatographic and spectroscopic means. Using xylitol and methylparaben as model reactants, three unknown peaks having the relation 1:2:4 were obtained together with the hydrolysis product 4-hydroxybenzoic acid. Diode array detection gave identical UV spectra for each peak with a maximum at 255 nm. The structures of the isomeric 1-, 2-, and 3-xylityl 4-hydroxybenzoic acid esters were proved by means of LC-MS, GC-MS, and NMR and correlated to the peaks in the HPLC chromatograms. The rate of the transesterification was shown to be highest in strongly alkaline medium (ph 10-11), whereas equilibration of the reaction was optimally balanced at pH 8-9. An increase of polyol concentration enhanced the formation of the esters. The reactivity of different substituted parabens was higher in the case of parabens with a short alkyl ester function. Similar reaction profiles were observed with C3-C6 polyols, but no transesterification took place when aldoses were used.

Chromatography, High Pressure Liquid↗
PubMed1995

Lymphocyte subsets in bronchoalveolar lavage after exposure to Actinobacillus pleuropneumoniae in pigs previously immunized orally or by aerosol.

Young pigs were immunized with the lung-pathogenic bacterium Actinobacillus (Haemophilus) pleuropneumoniae by aerosol or orally using viable and inactivated bacteria. The cellular changes in the bronchoalveolar lavage (BAL) were studied in repeated lavages after the pigs were infected with live bacteria. The nucleated cells in the BAL were differentiated and lymphocyte subsets determined. There were no major differences between the two routes of immunization or between viable and inactivated bacteria. The immunization induced an increase in all lymphocyte subsets studied and in the appearance of plasma cells and lymphoid blasts. The infection did not cause a further increase except in granulocytes. The lack of a booster-type increase in lymphocytes in the BAL might indicate a different immunologic reaction of the lung or that lymphocytes of the BAL do not represent lung lymphocytes in general. The protective effect of the immunization might be deduced from the increase in lymphocytes after immunization but not from the reaction pattern after infection.

Actinobacillus Infections↗
PubMed1995

gamma-Propoxy-sulfo-lichenin, an antitumor polysaccharide derived from lichenin.

A water-soluble semisynthetic polysaccharide, gamma-propoxy-sulfo-lichenin (PSL), was prepared by reaction of propansultone with lichenin, a natural occurring beta-1.3/1.4-linked glucan originating from Cetraria sp. PSL represents a class of mixed-linked beta-glucans with long and hydrophilic side chains in position C-6 of the glucan backbone. PSL with a degree of substitution of 0.8 and an average molecular weight of 250 kDa exhibited a strong antitumor activity in doses of 25 and 5 mg/kg against solid sarcoma 180 (100% resp. 82% tumor inhibition). The antitumor activity of PSL was shown to be dependent on the dimension of the molecule: the higher the average molecular weight, the higher was the inhibition rate obtained in the antitumor assay. No antitumor effect was observed by using a pretreatment of animals prior to transplantation of sarcoma 180. On syngenic DBA/2-MC.SC1 fibrosarcoma PSL inhibited tumor growth by about 88% at a concentration of 25 mg/kg. PSL failed to exhibit any direct cytotoxic effects on hormone-independent MDA-MB 231 mammacarcinoma. For PSL an indirect antitumor effect via modulation of the host mediate immune defence is postulated.

Animals↗
PubMed1995

Aerosol exposure of pigs to viable or inactivated Actinobacillus pleuropneumoniae serotype 9 induces antibodies in bronchoalveolar lining fluids and serum, and protects against homologous challenge.

A dose-defined nose-only inhalation system for pigs was used to study the immunogenic and protective potentials of a single aerosol application of viable or killed Actinobacillus pleuropneumoniae serotype 9. Respiratory volumes were measured for each pig to calculate inhaled individual doses. Eight pigs inhaled 107 CFU A. pleuropneumoniae CVI 13261 reference strain for serotype 9. Another eight pigs received an identical dose of killed actinobacilli. After three weeks the pigs and nonexposed controls were challenged with 108 CFU of the homologous strain by aerosol. Bronchoalveolar lavage (BALF) in pigs was performed during the experiment to obtain lavage samples for assessment of local antibodies. Isotype-specific antibody responses in serum and BAL fluids were measured by ELISAs based on whole-cell antigens. The protective efficacy of aerosol immunization was evaluated by clinical and post-mortem examinations. The controls developed fever and severe pleuropneumonia, whereas previously exposed pigs had less fever and less extensive gross pulmonary lesions. After the first aerosol exposure pulmonary IgM, and IgG antibodies reactive with A. pleuropneumoniae increased significantly in both aerosol exposed groups. IgA in BALF and serum concentrations of each Ig class were significantly increased in the group exposed to viable bacteria when compared to the non-exposed controls. After aerosol challenge a pronounced increase of systemic and pulmonary IgA, IgM, and IgG antibodies was detected in both exposure groups. Aerosol application of whole-cell A. pleuropneumoniae bacterins induced similar protective effects against aerosol challenge infection as administration of an identical dose of viable bacteria. Inhalation of A. pleuropneumoniae may lead to asymptomatic carriers in some pigs that could spread the disease under field conditions.

Actinobacillus Infections↗
PubMed1995

Oral immunization of pigs with viable or inactivated Actinobacillus pleuropneumoniae serotype 9 induces pulmonary and systemic antibodies and protects against homologous aerosol challenge.

A dose-defined aerosol infection of pigs was used to study the immunogenic and protective potentials of oral immunization with dead or live Actinobacillus pleuropneumoniae serotype 9 reference strain CVI 13261 against an aerogenic challenge. Pigs were vaccinated with a single dose of 10(11) CFU of viable (n = 8) or inactivated (n = 8) A. pleuropneumoniae given orally in a gelatin capsule. After 3 weeks, vaccinated pigs and nonvaccinated controls were challenged aerogenically with a dose of 10(8) CFU of A. pleuropneumoniae CVI 13261. The protective efficacy of oral immunization was evaluated by clinical and postmortem examinations. Bronchoalveolar lavage in pigs was performed during the experiment to obtain lavage samples for assessment of local antibodies. Isotype-specific antibody responses in sera and in bronchoalveolar lavage fluids were determined by enzyme-linked immunosorbent assays based on whole-cell antigen. Oral immunization did not induce clinical side effects. After aerosol challenge, two animals of both vaccinated groups (25% in each case) showed a moderate fever for 2 days, whereas all four pigs (100%) of the nonvaccinated control group developed severe fever. In contrast to the controls, which developed severe pleuropneumonia, the vaccinated pigs had only mild pulmonary lesions. Three weeks after challenge, 13 of 16 vaccinated pigs (81%) were found to be free of pathomorphological changes of the lungs. From two of these pigs immunized with live bacteria we were able to reisolate A. pleuropneumoniae. A significant systemic and pulmonary increase in the concentrations of immunoglobulin A (IgA), IgM, and IgG antibodies reactive with A. pleuropneumoniae was detectable after aerosol challenge in both vaccinated groups. Immunization with viable bacteria was found to induce significantly higher concentrations of each Ig isotype in bronchoalveolar lavage fluids and sera than immunization with inactivated A. pleuropneumoniae. These serological findings were not reflected in the reduction in clinical disease after challenge in comparison to the case for the pigs vaccinated with inactivated bacteria. We concluded that a single oral administration of A. pleuropneumoniae provides partial clinical protection against aerosol challenge infection in the respiratory tract.

Actinobacillus Infections↗
PubMed1994

Immunogenicity of Vibrio cholerae ghosts following intraperitoneal immunization of mice.

The immunogenic potential of Vibrio cholerae ghosts (VCG) in comparison with heat-killed whole-cell vibrios (WCV) was evaluated after intraperitoneal immunization of adult mice. Swiss white mice received four doses of VCG or WCV intraperitoneally, consisting of 500 micrograms of lyophilized material in 200 microliters of phosphate-buffered saline (PBS), pH 7.4. The control group received 200 microliters of PBS. Serum samples were collected from all mice on the day of immunization and on days 14, 24, 35 and 62 postimmunization. Sera were examined for vibriocidal antibodies by the microtitre and tube-dilution methods and Vibrio-specific serum IgG antibodies were assessed by ELISA. IgG antibodies to intact WCV were detected in sera from all animals immunized with VCG or WCV. The response was specific and of high magnitude. Significantly higher antibody responses were obtained when sera from both VCG- and WCV-immunized mice were titrated against VCG. The immunogenicity of VCG in evoking serum IgG responses was higher than that of WCV. However, the immunogenicity of the two antigen preparations was comparable in terms of seroconversion for vibriocidal antibodies. These results demonstrate that VCG administered intraperitoneally evoke Vibrio-specific serum IgG responses as well as vibriocidal antibody activity in mice.

Animals↗
PubMed1994

Oral and aerosol immunization with viable or inactivated Actinobacillus pleuropneumoniae bacteria: antibody response to capsular polysaccharides in bronchoalveolar lavage fluids (BALF) and sera of pigs.

To investigate the antibody response after local application of lung-pathogenic bacteria, pigs were immunized with viable or inactivated Actinobacillus pleuropneumoniae by the oral and aerogenous route. After 3 weeks class-specific immunoglobulins against purified A. pleuropneumoniae capsular polysaccharides (CP) were determined in serum and BALF by ELISA. A significant increase of IgA antibodies was found in BALF but not in sera of all immunized pigs. Oral immunization with viable A. pleuropneumoniae and aerosol immunization with either viable or inactivated bacteria resulted in a significant increase of IgG antibodies to the CP antigen in BALF, whereas only aerosol exposure to viable bacteria resulted in a significant increase in IgG antibodies in serum. A significant increase in anti-CP IgM in BALF was observed after aerosol exposure but not after oral immunization. IgM antibodies towards CP increased significantly by both routes of immunization with viable bacteria. The anti-CP activity of all three isotypes in sera and BALF was low in all groups compared with the positive controls, although inoculation of viable A. pleuropneumoniae led to higher levels of antibody concentration than inactivated bacteria. Our results indicate a traffic of primed lymphocytes from the gut into the bronchoalveolar airways and further support the hypothesis that polysaccharide-specific B cells may functionally mature at the mucosal surfaces.

Actinobacillus pleuropneumoniae↗
PubMed1994

Influence of Serum and Glucose Additives on Survival of Actinobacillus pleuropneumoniae Aerosolized from the Freeze-Dried State.

Serum and/or glucose added to Actinobacillus pleuropneumoniae suspensions before freeze-drying significantly increased survival rates of bacteria in aerosols. Aerosols with predictable numbers of viable bacteria can be made as required in an aerosol infection model. Sucrose supplementation of impinger fluids increased recovery of viable A. pleuropneumoniae.

Journal Article↗
PubMed1994

Prevalence of aerobic bacteria in bronchoalveolar lavage fluids from healthy pigs.

Fiberoptic bronchoscopy was performed in pigs to assess bacterial contamination of bronchoalveolar lavage fluids (BALF) obtained by use of the method and to determine the aerobic bacterial species in bronchoalveolar airways of healthy pigs. Bacterial contamination of BALF caused by insertion of the bronchoscope was evaluated, using a chromogenic bacterial tracer strain, and was found to be 0.22% of total colony-forming units (CFU), with range between 0 and 1.6%. A total of 164 pulmonary-healthy pigs from 6 closed herds were selected. The BALF obtained from these pigs were examined bacteriologically. Bacteria could not be isolated from 10.4% of all BALF; 5.5% of the BALF samples yielded pure cultures; and 84.1% yielded mixed aerobic bacterial growth. In BALF from 29.2% of the pigs, < or = 5 x 10(2) CFU of bacteria/ml were isolated. The total number of bacteria in BALF from 50% of the pigs varied between 5 x 10(2) and 10(3) CFU/ml; 10.4% of BALF samples contained between 10(3) CFU/ml and 5 x 10(3) CFU/ml. More than 1 bacterial species were isolated from a single lung lavage of 84.1% of the pigs. Up to 6 species were isolated from a single BALF sample. A total of 443 bacterial isolates were differentiated into 25 bacterial genera and species. Samples of BALF yielded staphylococci (67.6%: Staphylococcus hyicus from 13.4% of the samples and S aureus from 2.4%), alpha-hemolytic streptococci (49.4%), Escherichia coli (42.1%), non-hemolytic streptococci (26.2%), Klebsiella spp (18.3%), micrococci (12.8%), and Coryneformes (11.0%). Other bacterial species were found, but less frequently.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Cellular changes in the bronchoalveolar lavage (BAL) of pigs, following immunization by the enteral or respiratory route.

Normal young pigs were immunized by the oral or aerogenic route with the viable or inactivated lung-pathogenic bacterium Actinobacillus (Haemophilus) pleuropneumoniae. Three weeks later the cellular composition as well as the lymphocyte subset composition of the bronchoalveolar space were examined by BAL. Lymphocytes in the lavage increased significantly, including CD4+ and CD8+ T cells. After oral immunization a dramatic increase of plasma cells and lymphoid blasts was found. Among immunoglobulin-positive lymphocytes IgG+ cells showed the most pronounced increase. For most lymphocyte subsets there was no difference between viable and inactivated bacteria. Oral immunization with a lung-pathogenic bacterium results in increased numbers of lymphocytes in the bronchoalveolar space and might play a critical role in protection against lower respiratory tract infections.

Actinobacillus pleuropneumoniae↗
PubMed1992

Effects of microbial stimulation on the number, size and activity of bronchus-associated lymphoid tissue (BALT) structures in the pig.

The development of bronchus-associated lymphoid tissue (BALT) was investigated in the pig, which is a species in which BALT is not found constantly. Different routes of contact with a specifically lung-pathogen bacterium Actinobacillus (Haemophilus) pleuropneumoniae were tested. Pigs, selected by bacteriological screening methods and the number of granulocytes in the bronchoalveolar lavage (BAL), were infected by aerosol. They were compared to previously enterally immunized pigs using active and inactivated bacteria. The development of BALT after the infection was compared to that in pigs with a single enteral, or no, contact with the bacterium. BALT was less frequent in these groups than in the infected pigs. Previously immunized pigs developed the highest number and the largest BALT with the most prominent morphological signs of activation. Immunization with living or inactivated bacteria did not cause histological differences. BALT was preferentially located around bronchioli and small bronchi. Additional BALT predominantly occurred in the walls of larger bronchi. Definite compartments of T and B lymphocytes were not found in immunohistological studies of BALT. It was concluded that the development of BALT can be induced by different modes of microbial stimulation.

Actinobacillus Infections↗
PubMed1991

Solubilization and properties of GDP-fucose: xyloglucan 1,2-alpha-L-fucosyltransferase from pea epicotyl membranes.

GDP-fucose:xyloglucan 1,2-alpha-L-fucosyltransferase from pea (Pisum sativum) epicotyl microsomal membranes was readily solubilized by extraction with the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (Chaps). When using GDP-[14C]fucose as fucosyl donor and tamarind xyloglucan (XG) as acceptor, maximum activation was observed at 0.3% (w/v) Chaps and the highest yield of solubilized activity at 0.4%. The reaction product was hydrolyzed by Trichoderma cellulase to yield labeled oligosaccharides that peaked on gel permeation chromatography at the same elution volume as pea XG nona- and decasaccharide subunits. The apparent Km for fucosyl transfer to tamarind XG by the membrane-bound or solubilized enzyme was about 80 microM GDP-fucose. This was 10 times the apparent Km for fucosyl transfer to endogenous pea nascent XG. Optimum activity was between pH 6 and 7, and the isoelectric point was close to pH 4.8. The solubilized enzyme showed no requirement for, or stimulation by, added cations or phospholipids, and was stable for several months at -70 degrees C. Solubilization and gel permeation chromatography on columns of Sepharose CL-6B enriched the specific activity of the enzyme by about 20-fold relative to microsomes. Activity fractionated on columns of CL-6B with an apparent molecular weight of 150 kDa. The solubilized fucosyltransferase was electrophoresed on nondenaturing polyacrylamide slab gels containing 0.02% (w/v) tamarind XG, and its activity located by incubation in GDP-[14C]fucose, washing, and autoradiographing the gel. A single band of labeled reaction product appeared with an apparent molecular weight of 150 kDa.

Chromatography, Gel↗
PubMed1990

Yeast KRE genes provide evidence for a pathway of cell wall beta-glucan assembly.

The Saccharomyces cerevisiae KRE1 gene encodes a Ser/Thr-rich protein, that is directed into the yeast secretory pathway, where it is highly modified, probably through addition of O-linked mannose residues. Gene disruption of the KRE1 locus leads to a 40% reduced level of cell wall (1----6)-beta-glucan. Structural analysis of the (1----6)-beta-glucan fraction, isolated from a strain with a krel disruption mutation, showed that it had an altered structure with a smaller average polymer size. Mutations in two other loci, KRE5 and KRE6 also lead to a defect in cell wall (1----6)-beta-glucan production and appear to be epistatic to KRE1. These findings outline a possible pathway of assembly of yeast cell wall (1----6)-beta-glucan.

Amino Acid Sequence↗