Lymphoid tissue-resident Alcaligenes LPS induces IgA production without excessive inflammatory responses via weak TLR4 agonist activity.

Alcaligenes are opportunistic commensal bacteria that reside in gut-associated lymphoid tissues such as Peyer's patches (PPs); however, how they create and maintain their homeostatic environment, without inducing an excessive inflammatory response remained unclear. We show here that Alcaligenes-derived lipopolysaccharide (Alcaligenes LPS) acts as a weak agonist of toll-like receptor 4 and promotes IL-6 production from dendritic cells, which consequently enhances IgA production. The inflammatory activity of Alcaligenes LPS was weaker than that of Escherichia coli-derived LPS and therefore no excessive inflammation was induced by Alcaligenes LPS in vitro or in vivo. Alcaligenes LPS also showed adjuvanticity, inducing antigen-specific immune responses without excessive inflammation. These findings reveal the presence of commensal bacteria-mediated homeostatic inflammatory conditions within PPs that produce optimal IgA induction without causing pathogenic inflammation and suggest that Alcaligenes LPS could be a safe and potent adjuvant.

Innate lymphoid cells promote anatomical containment of lymphoid-resident commensal bacteria.

The mammalian intestinal tract is colonized by trillions of beneficial commensal bacteria that are anatomically restricted to specific niches. However, the mechanisms that regulate anatomical containment remain unclear. Here, we show that interleukin-22 (IL-22)-producing innate lymphoid cells (ILCs) are present in intestinal tissues of healthy mammals. Depletion of ILCs resulted in peripheral dissemination of commensal bacteria and systemic inflammation, which was prevented by administration of IL-22. Disseminating bacteria were identified as Alcaligenes species originating from host lymphoid tissues. Alcaligenes was sufficient to promote systemic inflammation after ILC depletion in mice, and Alcaligenes-specific systemic immune responses were associated with Crohn's disease and progressive hepatitis C virus infection in patients. Collectively, these data indicate that ILCs regulate selective containment of lymphoid-resident bacteria to prevent systemic inflammation associated with chronic diseases.

Antigenic determinants of the membrane-bound hydrogenase in Alcaligenes eutrophus are exposed toward the periplasm.

Electron microscopic immunogold labeling experiments were performed with ultrathin sections of plasmolyzed cells of Alcaligenes eutrophus and "whole-mount" samples of spheroplasts and protoplasts. They demonstrated that antigenic determinants of the membrane-bound hydrogenase are exposed, at the outside of the cytoplasmic membrane, to the periplasm.

A monoclonal antibody-based latex bead agglutination test for the detection of Bordetella avium.

The purpose of this study was to develop a rapid method to distinguish Bordetella avium from closely related Bordetella avium-like and B. bronchiseptica bacteria. A monoclonal antibody of the IgM isotype was produced in Balb/c mice against live B. avium strain 75. The monoclonal antibody, in the form of ascites fluid, was added to a bovine serum albumin-glycine buffer (pH 8.6) and adsorbed to 3.03-microns-diameter latex beads. Optimum concentrations of antibody, beads, and bacteria were determined. The latex bead conjugate was tested against 40 isolates of B. avium, 24 isolates of B. avium-like bacteria, 17 isolates of B. bronchiseptica, two isolates of Alcaligenes faecalis, and several other common genera. Strong agglutination occurred with all B. avium isolates and the two isolates of A. faecalis. Weak agglutination occurred with Staphylococcus aureus and two isolates of B. bronchiseptica. There was no agglutination with any of the B. avium-like isolates. The latex bead agglutination test may be useful as an aid in the identification of B. avium when used in conjunction with other criteria.

[Effects of indices of the test-system "age--normal antibodies--leukocytes--lymphocytes" on immunological parameters under the action of cancer risk factors]. / Vliianie pokazatelei test-sistemy "vozrast--normal'nye antitela-- leikotsity--limfotsity" na immunologicheskie parametry organizma pri deistvii faktora onkologichedskogo riska.

It is established that application of the test-system "age-normal antibodies-leucocytes-lymphocytes" permits refusing in certain cases from the generally-accepted laboratory immunological tests and prognosticate their values depending on the test-system indices when using the regression models. The great incongruity of the predicted and real values of the concrete immunological index may be the manifestation of the risk factors associated with the oncological and other diseases.

Purification and properties of a protein linked to the soluble hydrogenase of hydrogen-oxidizing bacteria.

In Alcaligenes eutrophus, the formation of the hydrogenases and of five new peptides is subject to the hydrogenase control system. Of these, the B peptide was purified to homogeneity. This protein (Mr, 37,500) was composed of two identical subunits (Mr, 18,800). Antibodies against the B protein were used for its quantification by rocket immunoelectrophoresis. About 4% of the total protein consisted of the B protein; its molar ratio to the NAD-linked hydrogenase was about 4:1. The B protein appeared to be associated with the NAD-linked hydrogenase, as shown by gel filtration analysis with Sephadex G-200. The B protein was not detected in cells that had not expressed the hydrogenase proteins or that lacked the genetic information of the hydrogen-oxidizing character; it was also not detected in Tn5 insertional mutants that were unable to form soluble hydrogenase antigens. Immunochemical analysis of other species and genera than A. eutrophus revealed that only strains able to form a NAD-linked hydrogenase also formed B-protein antigens. The B protein is not required for the catalytic activity of soluble hydrogenase in vitro; its function is at present unknown.

[Antigenic polysaccharides of bacteria. 19. The structure of the O-specific polysaccharide chain of Alcaligenes faecalis lipopolysaccharide]. / Antigennye polisakharidy bakterii. 19. Stroenie O-spetsificheskoi polisakharidnoi tsepi lipopolisakharida Alcaligenes faecalis.

On mild acid hydrolysis of Alcaligenes faecalis lipopolysaccharide, the O-specific polysaccharide containing D-rhamnose and D-xylose in the 3:2 ratio was obtained. Solvolysis of the polysaccharide with hydrogen fluoride in methanol resulted in methyl glycoside of a branched tetrasaccharide including three rhamnose and one xylose residues. Smith degradation of the polysaccharide led to the glycoside of disaccharide composed of two rhamnose residues and glycerol. On the basis of identification of the oligosaccharide fragments, methylation, 1H and 13C NMR analysis (including nuclear Overhauser effect data), it was established that the polysaccharide linear chain is a rhamnan, both xylose residues being attached to one of the rhamnose residues as two branches. The repeating unit of the polysaccharide has the following structure: (Formula: see text).

[Physico-chemical properties and heterogeneity of Alcaligenes faecalis polysaccharide]. / Fiziko-khimicheskie svoistva i geterogennost' lipopolisakharida Alcaligenes faecalis.

Two species of O-antigenic molecules with following sedimentation characteristics S(o)20, W 1,25.10(-13) S, D(o)20, W 9,7.10(-7) cm2/s, M 8000 and S(o)20, W 2,5.10(-13) S, D(o)20, W 5.10(-7) cm2/s, M 23,000-30,000 were detected in the cell wall of the strain Alcaligenes faecalis, a representative species of conditionally pathogenic microorganisms with unidentified taxonomic position. "Light" and "heavy" types of molecules have a lipopolysaccharide nature and show no differences in the monosaccharide composition of the polysaccharide moiety, structural organization of O-chain, or lipid A fatty-acid composition.

The efficacy of a temperature-sensitive mutant vaccine against Northwest Arkansas isolates of Alcaligenes faecalis.

The efficacy of a commercially produced temperature-sensitive mutant Alcaligenes faecalis vaccine was evaluated in turkeys contact-challenged with one of three strains of A. faecalis. In the vaccinated control group, the vaccine strain of A. faecalis colonized the nasal turbinates but not the trachea, caused no clinical signs of turkey coryza, and induced humoral antibodies. In the vaccinated challenged groups, the vaccine reduced both the severity of lesions and the number of birds exhibiting lesions compared with unvaccinated challenged groups, but it did not prevent colonization of challenge strains of A. faecalis.

Immune response of poults following intranasal inoculation with Artvax vaccine and a formalin-inactivated Bordetella avium bacterin.

Poults 3 weeks and older developed temporary tracheal resistance to intranasal challenge following inoculation of either Artvax vaccine or formalin-inactivated Bordetella avium bacterin by the intranasal and eyedrop routes. Resistance usually persisted for 3-4 weeks after B. avium challenge. However, with constant exposure to infected controls, the vaccinated birds eventually developed tracheal infection. Day-old poults did not respond to either the Artvax or the bacterin and were completely susceptible to challenge. Two-week-old poults responded to some degree, but poults 3 weeks old and older responded best. Poults inoculated with bacterin by aerosol or by drinking water did not respond as well as those inoculated by the intranasal and eyedrop routes. When poults were given a single subcutaneous injection at 3 weeks of age and challenged 2 weeks later, three of five resisted infection for 18 days.

Evidence for adherence-dependent cytotoxicity of Alcaligenes faecalis in turkey tracheal organ cultures.

A virulent isolate of Alcaligenes faecalis was examined in turkey tracheal organ cultures (TTOC) for adherence using immunofluorescent staining and for cytotoxicity using light microscopic observation. Treatment of the bacterial culture with trypsin, antiserum specific for A. faecalis, and N-acetylneuraminic acid inhibited the ability of the organism to adhere to TTOC. Treatment of the bacterium with D-galactose partly decreased adherence of the bacterium. Those treatments that inhibited the adherence of A. faecalis also inhibited the cytolytic activity in TTOC. Treatment of the bacterial culture with D-galactose only partly decreased the cytolytic activity. These data indicate that adherence of the organism to TTOC is necessary for the cytolytic activity characteristic of A. faecalis isolates capable of causing alcaligenes rhinotracheitis in turkeys.

Partial characterization of the hemagglutinin of Alcaligenes faecalis.

The hemagglutinin of Alcaligenes faecalis was partially characterized. Hemagglutination (HA) was blocked by enzymes inactivating proteins, by heat, and by antisera but not by sugar-blocking substances. Pili were not determined to be a factor in HA activity. There was no connection between virulence and HA activity.

In vitro cytotoxicity of an Alcaligenes faecalis and its relationship in in vivo tracheal pathologic changes in turkeys.

Two isolates of Alcaligenes faecalis from turkeys with respiratory disease were indistinguishable physically, biochemically, and for specific agglutinating antibodies. The isolates differed in in vitro cytotoxicity for turkey tracheal organ cultures and in ability to induce clinical rhinotracheitis in poults. The isolate designated NCDp induced in vitro cytotoxic changes in turkey tracheal organ cultures. Additionally, poults inoculated with NCDp developed severe clinical signs of rhinotracheitis, flaccid (collapsing) trachea, bacterial colonization of the cilia, and degeneration and loss of the columnar epithelium from the anterior one third to one half of the trachea. The isolate designated NCDm induced little or no cytotoxic changes in turkey tracheal organ culture. Isolate NCDm caused mild clinical signs of rhinotracheitis and colonized the trachea of inoculated poults, but it caused no other observable changes. A correlation seems to exist between in vitro cytotoxicity and in vivo pathogenicity for these isolates of A. faecalis.

Immunosuppressive properties and circulating life of Achromobacter glutaminase-asparaginase covalently attached to polyethylene glycol in man.

The immunosuppressive effects and circulating life of Achromobacter glutaminase-asparaginase (GA) covalently attached to polyethylene glycol (PEG) were examined in human subjects following a single iv dose of 1000 IU/m2. Plasma half-life of PEG-GA was 72 hours. Skin test reactivity to recall antigens (mumps and tuberculin) was lost in all four patients tested. In vitro phytohemagglutinin-induced blastogenesis, "natural killing," and phytohemagglutinin-induced cell cytotoxicity was diminished as long as enzyme levels were detectable. In vivo and in vitro activities returned to normal following total plasma clearance of enzyme.

Effect of humidity on infection of turkeys with Alcaligenes faecalis.

Turkeys maintained at 75% to 80% relative humidity were more adversely affected by Alcaligenes faecalis infection than turkeys maintained at 20 to 35% relative humidity. Alcaligenes faecalis was reisolated earlier and more often from turkeys maintained at the higher humidity. Clinically, the turkeys maintained at high humidity exhibited both sinusitis and conjunctivitis earlier than the turkeys at low humidity. In both groups, antibody titers as determined by a microagglutination test developed by 2 weeks postinoculation and started to decline after the third week, lymphocytosis was demonstrated at 1 week postinoculation, and a lymphopenia developed at 5 weeks postinoculation.

Control of turkey Alcaligenes rhinotracheitis in Utah with a live vaccine.

During the 1980 growing season in the Sanpete Valley of Utah, about half of the 250 to 300 flocks of turkeys were vaccinated with an oral vaccine against alcaligenes rhinotracheitis (ART). The vaccine consisted of a temperature-sensitive mutant of Alcaligenes faecalis. Most vaccinated birds developed serum antibodies. No outbreaks of ART occurred in vaccinated flocks, although some outbreaks occurred in unvaccinated flocks. During 1979, when on flocks were vaccinated, over 90% of the flocks experienced outbreaks of ART during late summer.

Negative findings concerning Alcaligenes faecalis as an etiologic agent in acute respiratory disease of turkeys.

An acute respiratory disease of turkeys in Israel was first reported in November 1978. Alcaligenes faecalis was isolated from sick turkeys and from chickens not affected by the disease. Plate agglutination tests with A. faecalis antigen of 1,067 turkey and 494 chicken serum samples gave variable results: healthy turkeys gave positive reactions and sick turkeys sometimes gave negative ones. All isolated strains were highly sensitive in vitro drug sensitivity tests, but chemotherapy failed in the field. Pathogenicity trials with A. faecalis, given alone or in combination with Yucaipa virus to 8-day-old turkey poults, failed to reproduce the disease.