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.

Community-acquired bacteremic Achromobacter xylosoxidans type IIIa pneumonia in a patient with idiopathic IgM deficiency.

A man was hospitalized with bacteremic Achromobacter xylosoxidans type IIIa pneumonia. The authors are aware of no previously reported similar infections caused by this bacterium. A clinical cure was achieved with a combination of carbenicillin and kanamycin therapy. Microtiter susceptibility testing revealed that carbenicillin was the antibiotic to which A. xylosoxidans IIIa was most sensitive (minimal inhibitory concentration, 1.6 microgram/ml) and that synergy between carbenicillin and kanamycin existed. During the patient's hospitalization, deficiency of IgM (21 mg/dl) was found. Specific serum activity against A. xylosoxidans IIIa was detected by the agglutination method. Specific anti-A. xylosoxidans IIIa IgG, but not IgM, was detected by indirect immunofluorescence. It appears that a defect in immunologic recognition of A. xylosoxidans IIIa as an invasive bacterium, a defect in synthesis of specific IgM, or both, contributed to this patient's infection.

Acinetobacter and similar organisms in ear infections.

Fifty-seven strains of acinetobacter-like organisms were isolated over a period of 26 months from the ears of 55 patients with acute or chronic otitis media, or otitis externa, and one strain was isolated in a survey of 50 normal ears. After comparison with eight reference strains, 32 of the isolates were identified as Acinetobacter anitratus, 22 as Acinetobacter Iwoffii, three as Moraxella spp. and one as Achromobacter sp. Analysis of the clinical findings suggests that although most of these organisms played little part in the disease process, a few strains were probably pathogenic in this situation.

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.

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.

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.

Field vaccination trials against turkey coryza using a temperature-sensitive mutant of Alcaligenes faecalis.

In field trials, 14 flocks of turkeys were vaccinated against coryza (rhinotracheitis) with a temperature-sensitive mutant of Alcaligenes faecalis via drinking water. Vaccine doses of 5 to 250 million bacteria per turkey were used. Turkeys ranged in age from 2.5 to 13 weeks. Doses of 5 to 7 million bacteria did not induce complete protection. Doses of 50 million bacteria or more appeared to induce substantial immunity in most flocks. Six-week-old birds receiving doses of 90 million bacteria developed a high degree of protection against challenge with virulent A. faecalis.

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.

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.

Development and use of amicroagglutination test to detect antibodies to Alcaligenes faecalis in turkeys.

A neotetrazolium-chloride-stained Alcaligenes faecalis antigen was developed for use in the microagglutination (MA) test. The test was used to detect serum antibodies in naturally and experimentally infected turkeys. The highest titer observed in naturally infected birds was 1:320. In one commercial flock, antibodies were detected at 12 and 15 weeks after the initial disease outbreak. Four experiments were conducted to study the serologic responses of turkeys to A. faecalis. Antibodies were first detected at 2 weeks postexposure (PE) in poults that were exposed to the organism at 1 week of age. Peak antibody titers were detected at 3 weeks PE; isolations of the organism then declined. No antibodies were detected at 7 weeks PE in these birds. Birds infected at 5 weeks of age via various routes developed maximum antibody titers 2 weeks PE. Birds inoculated subcutaneously had the highest titers, whereas those inoculated intramuscularly had the lowest titers. Antibodies were still detected at 56 days PE in some birds. Hens vaccinated with an inactivated A. faecalis bacterin developed antibody titers. Titers not higher than 1:40 were detected at hatching in progeny of these hens. However, these poults were not protected from disease after challenge. There was some evidence that birds exposed to live or inactivated A. faecalis develop some protection against challenge. Antigens were prepared using 4 Ohio A. faecalis isolates (A, B, C, and D) and 1 North Carolina isolate for use in the MA test. The results indicated that the 5 isolates were antigenically similar. Antigens prepared using isolate B reacted best in the MA test.

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.

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.

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.

Immunization against turkey coryza by colonization with mutants of Alcaligenes faecalis.

Temperature-sensitive mutants of Alcaligenes faecalis that exhibited reduced growth at 42 C but grew well at 30 C were induced by N-methyl-N'-nitro-N-nitrosoguanidine. Four of the 5 mutants that were studied readily colonized the nasal mucosa of turkeys when given intranasally or in drinking water. Several mutants caused no signs of coryza or depression and produced minimal colonization of the trachea. Colonization of the nasal mucosa lasted 3 to 4 weeks and induced moderate levels of serum antibodies. When challenged with virulent isolates of A. faecalis 4 to 6 weeks after having been exposed to the mutants, unstressed turkeys were protected; turkeys subjected to water deprivation exhibited some colonization with the virulent strain and showed slight signs of coryza.

[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.

[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.

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.