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Proteus penneri (P. penneri) is a bacillus-shaped, gram-negative, facultative anaerobe bacterium that is primarily an invasive pathogen and the etiological agent of several hospital-associated infections. P. penneri strains are naturally resistant to macrolides, amoxicillin, oxacillin, penicillin G, and cephalosporins; in addition, no vaccines are available against these strains. This warrants efforts to propose a theoretical based multi-epitope vaccine construct to prevent pathogen infections. In this research, reverse vaccinology bioinformatics and immunoinformatics approaches were adopted for vaccine target identification and construction of a multi-epitope vaccine. In the first phase, a core proteome dataset of the targeted pathogen was obtained using the NCBI database and subjected to bacterial pan-genome analysis using bacterial pan-genome analysis (BPGA) to predict core protein sequences which were then used to find good vaccine target candidates. This identified two proteins, Hcp family type VI secretion system effector and superoxide dismutase family protein, as promising vaccine targets. Afterward using the IEDB database, different B-cell and T-cell epitopes were predicted. A set of four epitopes "KGSVNVQDRE, NTGKLTGTR, IIHSDSWNER, and KDGKPVPALK" were chosen for the development of a multi-epitope vaccine construct. A 183 amino acid long vaccine design was built along with "EAAAK" and "GPGPG" linkers and a cholera toxin B-subunit adjuvant. The designed vaccine model comprised immunodominant, non-toxic, non-allergenic, and physicochemical stable epitopes. The model vaccine was docked with MHC-I, MHC-II, and TLR-4 immune cell receptors using the Cluspro2.0 web server. The binding energy score of the vaccine was - 654.7 kcal/mol for MHC-I, - 738.4 kcal/mol for MHC-II, and - 695.0 kcal/mol for TLR-4. A molecular dynamic simulation was done using AMBER v20 package for dynamic behavior in nanoseconds. Additionally, MM-PBSA binding free energy analysis was done to test intermolecular binding interactions between docked molecules. The MM-GBSA net binding energy score was - 148.00 kcal/mol, - 118.00 kcal/mol, and - 127.00 kcal/mol for vaccine with TLR-4, MHC-I, and MHC-II, respectively. Overall, these in silico-based predictions indicated that the vaccine is highly promising in terms of developing protective immunity against P. penneri. However, additional experimental validation is required to unveil the real immune response to the designed vaccine., Competing Interests: Declarations Conflict of Interest The authors declare no conflict of interest., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In the years 2006-2011, 617 Proteus spp. strains isolated mostly from urine and wounds or other clinical sources were collected in Łódź, Poland, to determine the offensive O serotypes frequently occurring among patients. P. mirabilis exhibited the most intensive swarming growth and was dominating species (86.9%), followed by P. genomospecies, P. vulgaris, and P. penneri. Ninety four per cent strains were recognized as S (smooth) forms. Serological studies (involving ELISA-enzyme-linked immunosorbent assay and Western blotting using native and adsorbed rabbit antisera) enabled classification of 80% S isolates into respective Proteus O serogroups among the 83 ones, described so far. The remaining strains seemed to be serologically unique. Despite the observed big serological variety of Proteus spp. isolates, we found the O78 serogroup recently described in Poland as dominating and identified other widespread serotypes: O3, O6, O10, O11, O27, O28, and O30 reported earlier as predominating also in other countries; O77 and O79 detected lately in Poland; O16, O18, O20, and O50. No unique structural feature of the prevalent O serotypes has been indicated. However, the prevalence of some O serogroups indicates that particular serotypes may be in some ways beneficial to the strains producing these kinds of O antigen., (© 2021. The Author(s).)

Proteus species are well-known opportunistic pathogens frequently associated with skin wound and urinary tract infections in humans and animals. O antigen diversity is important for bacteria to adapt to different hosts and environments, and has been used to identify serotypes of Proteus isolates. At present, 80 Proteus O-serotypes have been reported. Although the O antigen structures of most Proteus serotypes have been identified, the genetic features of these O antigens have not been well characterized. The O antigen gene clusters of Proteus species are located between the cpxA and secB genes. In this study, we identified 55 O antigen gene clusters of different Proteus serotypes. All clusters contain both the wzx and wzy genes and exhibit a high degree of heterogeneity. Potential functions of O antigen-related genes were proposed based on their similarity to genes in available databases. The O antigen gene clusters and structures were compared, and a number of glycosyltransferases were assigned to glycosidic linkages. In addition, an O serotype-specific suspension array was developed for detecting 31 Proteus serotypes frequently isolated from clinical specimens. To our knowledge, this is the first comprehensive report to describe the genetic features of Proteus O antigens and to develop a molecular technique to identify different Proteus serotypes.

We review the evidence that infectious agents, including those that become dormant within the host, have a major role to play in much of the etiology of rheumatoid arthritis and the inflammation that is its hallmark. This occurs in particular because they can produce cross-reactive (auto-)antigens, as well as potent inflammagens such as lipopolysaccharide that can themselves catalyze further inflammagenesis, including via β-amyloid formation. A series of observables coexist in many chronic, inflammatory diseases as well as rheumatoid arthritis. They include iron dysregulation, hypercoagulability, anomalous morphologies of host erythrocytes, and microparticle formation. Iron dysregulation may be responsible for the periodic regrowth and resuscitation of the dormant bacteria, with concomitant inflammagen production. The present systems biology analysis benefits from the philosophical idea of "coherence," that reflects the principle that if a series of ostensibly unrelated findings are brought together into a self-consistent narrative, that narrative is thereby strengthened. As such, we provide a coherent and testable narrative for the major involvement of (often dormant) bacteria in rheumatoid arthritis.

Proteus sp. and Klebsiella sp. mainly cause infections of the urinary and respiratory tracts or wounds in humans. The representatives of both genera produce virulence factors like lipopolysaccharide (LPS) or outer membrane proteins (OMPs) having much in common in the structures and/or functions. To check how far this similarity is revealed in the serological cross-reactivity, the bacterial masses of 24 tested Klebsiella sp. strains were tested in ELISA with polyclonal rabbit antisera specific to the representatives of 79 Proteus O serogroups. The strongest reacting systems were selected to Western blot, where the majority of Klebsiella masses reacted in a way characteristic for electrophoretic patterns of proteins. The strongest reactions were obtained for proteins of near 67 and 40 kDa and 12.5 kDa. Mass spectrometry analysis of the proteins samples of one Proteus sp. and one Klebsiella sp. strain showed the GroEL like protein of a sequence GI number 2980926 to be similar for both strains. In Western blot some Klebsiella sp. masses reacted similarly to the homologous Proteus LPSs. The LPS contribution in the observed reactions of the high molecular-mass LPS species was confirmed for Klebsiella oxytoca 0.062., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Proteus penneri, like the other seven species from the genus, are Gram-negative, peritrichously flagellated rods capable of swarming growth on humid solid media. These bacteria are human opportunistic pathogens involved in many infections but they mainly affect the urinary tract of hospitalized, long-term catheterized patients. P. penneri rods produce a lot of virulence factors, among which the lipopolysaccharide seems to be the most interesting due to its structural and serological diversity. From the three LPS regions of P. penneri strains only the core region and O-specific polysaccharide (OPS) were structurally and serologically examined. P. penneri LPS core region is characterized by a common inner part representing the III glycoform and a diverse distal part (12 different structures). The P. penneri O-antigens contain sugar and non-sugar compounds and some of them rarely occur in nature. In both P. penneri LPS regions putative epitopes have been pointed out. Serospecificity of OPS allowed classifying many P. penneri isolates to different Proteus sp. O-serogroups, among which 12 contain P. penneri strains only., (Copyright © 2013. Published by Elsevier Ltd.)

Lampreys, the surviving representative of jawless vertebrates, have been a focal point in the search for the evolutionary origin of adaptive immunity. They have independently evolved the variable lymphocyte receptor (VLR)-based adaptive immune system that protects themselves from infection by a variable of microorganisms. The standard immunization schedule for Japanese lamprey (Lampetra japonica) was established to prepare antisera by injection of Escherichia coli, Bacillus proteus, Staphylococcus aureus, Mycobacterium smegmatis, RRBCs, SRBCs, NB4 cells and Hela cells. In this study, we demonstrated the activities of lamprey antisera, which might be helpful to research the collaboration between VLR-based adaptive immune system and complement system in jawless vertebrates., (Copyright © 2012. Published by Elsevier Inc.)

This review is devoted to structural and serological characteristics of the O-antigens (O-polysaccharides) of the lipopolysaccharides of various Proteus species, which provide the basis for classifying Proteus strains to O-serogroups. The antigenic relationships of Proteus strains within and beyond the genus as well as their O-antigen-related bioactivities are also discussed.

Introduction: Both smooth and rough Proteus sp. strains can be found. The latter are characterized by their lack of an O-polysaccharide chain in the lipopolysaccharide (LPS) molecule, which makes them suitable for obtaining anti-core sera. Using this kind of material enables identifying fragments of the Proteus LPS core region that might be involved in cross-reactions. To date only a few similar epitopes have been established for the genus Proteus., Materials and Methods: Polyclonal rabbit antisera directed against three rough strains of Proteus sp. were tested by enzyme-linked immunosorbent assay (ELISA) with a set of LPSs. The reactivity of the selected cross-reactive and homologous systems was checked by the Western blot technique and by a passive immunohemolysis assay preceded by the absorption of each antiserum with appropriate cross-reactive and homologous alkalized LPSs., Results: On the basis of the ELISA results, 19 cross-reactive antigens were selected among which both smooth and rough LPS forms were found. All the observed reactions involved the core region of the LPS. Using the antisera absorbed with the appropriate LPSs allowed identification of four groups of antigens with serologically identical core regions., Conclusions: Comparing the results of the serological studies with the known chemical structures of the core regions of the LPSs used enabled the identification of a few core oligosaccharide fragments probably involved in the observed cross-reactions. All were located in the most distal part of LPS core region, which made them more easily recognized by specific antibodies.

Larrea divaricata is an abundant plant of northwest of Argentina used to treat different pathologies. We aimed to characterize the immunogenicity of proteins from a partially purified crude aqueous extract (JPCE) of jarilla. We evaluated the cross reaction between JPCE and whole cell-bacterial proteins (W-CBP) of Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris, and Klebsiella pneumoniae using a mouse anti-JPCE serum. Protein profiles of JPCE and W-CBP were analyzed. For JPCE, 18 bands were observed in a 20-176 kDa range. Levels of IgG against JPCE and W-CBP were determined. Bacterial proteins showed a strong reaction with the anti-JPCE serum. Plant proteins could be used as immune stimulants.

The serological cross-reactivity between lipopolysaccharides (LPS) of S. fidelis KMM3582(T) and rabbit anti-O P. mirabilis antibodies was tested. Using ELISA and Western blot cross-reactivity between S. fidelis LPS and antisera against P. mirabilis O14, O3 LPSs was found. The observed cross-reaction may suggest that anti-P. mirabilis S1959 (O3) antibodies may bind to the internal part of S. fidelis O-polysaccharides. A weak interaction between S. fidelis LPS and antiserum against P. mirabilis O13 in Western blot suggests that the absolute configuration of non-sugar "AlaLys" component (N(epsilon)-[(S)-l-carboxyethyl]-N(alpha)-(D-galacturonoyl)-L-lysine) may influence the affinity of antibodies for S. fidelis LPS.

Anti-tumor preventative effect of mono-therapy with the use of Proteus vaccine, Staphylococcus antitoxin and divaccine of Staphylococcus-Proteus has been studied. Experiments were carried out on 80 non-purebred laboratory white mice (age--3-3,5 months, body mass--18-20 g) and 60 rats (body mass--100-120 g) using intraperitoneal inoculation of Ehrlich's adenocarcinoma (ascitic form--EAT, in mice, cancer cells--3 x 10(6)), and subcutaneous inoculation of Sarcoma S-45 (in rats). Anti-tumor preventative effect of bacterial vaccines and immunization was evaluated according to the following parameters: Frequency of cancer development, Inhibition of cancer growth, Body mass index of experimental animals, Volume of ascitic fluid. Results of experiments have shown that use of bacterial polysaccharides with preventative purposes has better effect at S-45 growth than at EAT growth; Vaccination with Proteus prolongs lifespan mach more than vaccination with antitoxin of Staphylococcus; Vaccination with complex divaccine of Staphylococcus-Proteus causes complete resorption of tumors from 32 to 60 days; Development of experimental malignant tumors depends on type of anti-microbial vaccines and starting date of inoculation after completion of vaccination.

Bacterial strains have complex and individual antigenic structure, which provides basis for their serological identification. However, serological cross-reaction may occur when antibodies against a certain strain recognize other strains too. The molecular basis of this phenomenon is the expression of similar or identical antigenic epitopes on the surface of different bacterial cells. Such cross-reactions might harden the serological diagnosis of pathogenic bacteria. But it can be also advantageous, when antigens of non-pathogenic strains can be used in the serological examinations. Serological cross-reaction between three taxonomically different strains--Proteus morganii O34 (8662/64), Escherichia coli O111 and Salmonella Adelaide O35--have been described. It has been proven that it is based partially on the similar lipopolysaccharide structures of these pathogens. In this study the involvement of the outer membrane proteins of these strains in the serological cross-reaction is presented. Microfluidic chip technology was applied for the detection of common proteins, which provided fast and quantitative data about the proteins that might be responsible for serological cross-reaction. Two outer membrane proteins with apparent molecular mass of 36 and 41 kDa, respectively, could be detected in the profile of each strain, while individual dominating protein peaks have also appeared in the protein profiles. The presence of common protein antigens was proven by Western blotting.

Background: Both rheumatoid arthritis (RA) and ankylosing spondylitis (AS) are potentially disabling arthritic disorders for which as yet no highly sensitive and reliable diagnostic laboratory markers are available., Objective: The objective of this study was to evaluate the levels of antibodies against Proteus and Klebsiella antigenic peptides in an endeavor to develop diagnostic indices for the identification of patients with RA and AS, respectively., Methods: Sera from 50 patients with RA, 34 patients with AS, and 38 healthy subjects were screened for antibodies against "ESRRAL" and "IRRET" synthetic amino acid peptides obtained from Proteus hemolysin and urease (HU) as well as against "QTDRED" and "DRDE" peptides from Klebsiella nitrogenase and pullulanase (NP) proteins, respectively. Multiplication of the 2 antibodies against each organism produced indices for RA-HU and AS-NP., Results: Significantly increased levels of anti-HU antibodies (P<0.0001) were observed in patients with RA when compared with patients with AS or with healthy control subjects. Patients with AS were found to have significantly elevated levels of anti-NP (P<0.0001) antibodies when compared with patients with RA or with healthy subjects. Furthermore, all patients with RA were found to have values of anti-HU antibody (RA-HU) index above 95% confidence limit (CL) of the mean of healthy control subjects; meanwhile, all patients with AS were having values of anti-NP antibody (AS-NP) index above the 95% CL of the mean of healthy control subjects (100% sensitivity). However, the specificity of the RA-HU index in RA and the AS-NP index in patients with AS were 92% and 95%, respectively., Conclusion: The use of the RA-HU or AS-NP diagnostic index as a sole marker or in combination with other autoantibody markers could be used in the identification of patients with RA or AS, respectively. Longitudinal investigations starting with patients with early disease will be needed.

It has been suggested that Proteus infection may be involved in the pathogenesis of rheumatoid arthritis (RA). Bacterial and peptide immune responses in patients with RA and other control subjects were investigated in two geographically different populations. Serum samples from Finnish patients with early ( n=72) and advanced ( n=27) RA and 30 Finnish healthy controls, as well as from Japanese RA patients from two different locations: Tokyo ( n=30) and Otsu ( n=30), 18 patients with systemic lupus erythematosus (SLE) and 23 Japanese healthy controls were all screened for the total, and class-specific (IgG, IgA and IgM) antibodies against Proteus mirabilis, Escherichia coli and Serratia marcescens by indirect immunofluorescence assay. These samples were also tested for the determination of levels of isotypic antibodies against the shared epitope involving 16-mer synthetic peptides containing the EQRRAA or ESSRAL sequences and compared to scrambled control peptide by using an enzyme-labeled immunosorbent assay method. Significantly elevated levels of IgG and IgM antibodies to P. mirabilis and antibodies against both EQRRAA and ESSRAL peptides were detected in sera of Finnish patients with early and advanced RA, and in Japanese patients from Otsu or Tokyo compared to their corresponding control groups. In contrast, no difference either in the total or in any of the isotypic antibodies were observed between these groups when serum samples were screened against each of E. coli and S. marcescens or against the control peptide. Furthermore, there was a significant correlation between the antibody levels against Proteus bacteria only and both EQRRAA and ESRRAL peptides. Our findings support the possibility for specific involvement of P. mirabilis in the etiopathogenesis of RA even in early cases.

The lipopolysaccharides (LPS) of Proteus penneri 28 and Proteus vulgaris O31 (PrK 55/57) were degraded with dilute acetic acid and structurally identical high-molecular-mass O-polysaccharides were isolated by gel-permeation chromatography. Sugar analysis and nuclear magnetic resonance (NMR) spectroscopic studies showed that both polysaccharides contain D-GlcNAc, 2-acetamido-2,6-dideoxy-L-glucose (L-2-acetamido-2,6-dideoxyglucose (N-acetylquinovosamine)) and 2-acetamido-3-O-[(S)-1-carboxyethyl]-2-deoxy-D-glucose (N-acetylisomuramic acid) and have the following structure: [carbohydrate structure: see text] where (S)-1-carboxyethyl [a residue of (S)-lactic acid] (S-Lac) is an ether-linked residue of (S)-lactic acid. The O-polysaccharide studied is structurally similar to that of P. penneri 26, which differs only in the absence of S-Lac from the GlcNAc residue. Based on the O-polysaccharide structures and serological data of the LPS, it was suggested classifying these strains in one Proteus serogroup, O31, as two subgroups: O(31a), 31b for P. penneri 28 and P. vulgaris PrK 55/57 and O31a for P. penneri 26. A serological relatedness of the LPS of Proteus O(31a), 31b and P. penneri 62 was revealed and substantiated by sharing epitope O31b, which is associated with N-acetylisomuramic acid. It was suggested that a cross-reactivity of P. penneri 28 O-antiserum with the LPS of several other P. penneri strains is due to a common epitope(s) on the LPS core.

The influence of the newly developed complex vaccine Pyopol, containing the antigens of opportunistic microorganisms and polyoxydonium used as immunomodulator, on the oxygen-dependent metabolism of neutrophils was studied. The study revealed that the main components of the vaccine, both individually and in association, did not change cellular activity in the range of concentrations used in this study. The inhibition of the oxygen-dependent metabolism of neutrophil granulocytes in the presence of native or weakly diluted vaccine occurred due to the cytotoxic effect of thimerosal used as preservative.

The majority of anti-phosphocholine (PC) antibodies induced by the PC epitope in Proteus morganii (PM) express the M603 idiotype (id), which is characterized by an invariant Asp to Asn substitution at the V(H):D(H) junction. To elucidate the molecular basis by which M603-like B cells acquire the mutations resulting in this invariant substitution, we analyzed the immune response to PC-PM in terminal deoxynucleotidyl transferase (TdT) gene knockout (KO) mice. In the absence of TdT, T15-id antibodies comprised 80-100% of the primary response to PC-PM. Less than 10% of the response in wild-type mice is T15-id(+). In TdT KO mice, the secondary response to PC-KLH was higher than in wild-type mice and was dominated by the germ-line T15-id. About 10% of this response, in both TdT KO and wild-type mice, comprised M167-id(+) antibodies. Additionally, none of the functionally rearranged V1/DFL16.1/J(H)1 cDNA isolated from PC-PM-immunized TdT KO mice showed the Asp/Asn substitution characteristic of PC-binding, PC-PM-induced M603-like antibodies. These data indicate that production of M603-id antibody is TdT dependent, while generation of M167-id antibody is TdT independent, and that in the absence of competition from M603-like B cells, T15-id B cells can respond to PC-PM.

O-specific polysaccharides (O-antigens) of the lipopolysaccharides (LPS) of Proteus penneri strains 1 and 4 were studied using sugar analysis, (1)H and (13)C NMR spectroscopy, including 2D COSY, H-detected (1)H,(13)C HMQC, and rotating-frame NOE spectroscopy (ROESY). The following structures of the tetrasaccharide (strain 1) and pentasaccharide (strain 4) repeating units of the polysaccharides were established: [reaction: see text]. In the polysaccharide of P. penneri strain 4, glycosylation with the lateral Glc residue (75%) and O-acetylation of the lateral GalNAc residue (55%) are nonstoichiometric. This polysaccharide contains also other, minor O-acetyl groups, whose positions were not determined. The structural similarity of the O-specific polysaccharides was consistent with the close serological relatedness of the LPS, which was demonstrated by immunochemical studies with O-antisera against P. penneri 1 and 4. Based on these data, it was proposed to classify P. penneri strains 1 and 4 into a new Proteus serogroup, O72, as two subgroups, O72a and O72a,b, respectively. Serological cross-reactivity of P. penneri 1 O-antiserum with the LPS of P. penneri 40 and 41 was substantiated by the presence of an epitope(s) on the LPS core region shared by all P. penneri strains studied.

O-specific polysaccharide (O-antigen) of the lipopolysaccharide of Proteus penneri 20 was studied using sugar analysis along with various one- and two-dimensional NMR spectroscopy techniques. The following structure of the polysaccharide was established: [formula: see text] It has the same carbohydrate backbone structure as that described earlier for P. penneri 16, in which the positions of the O-acetyl groups have not been determined. P. penneri 20 O-antiserum showed a strong cross-reactivity with the lipopolysaccharides of P. penneri 10, 16, 18, 32, 45 and P. mirabilis O17. These data enable classifying these strains together with P. penneri 20 in one Proteus serogroup, O17.

A phosphorylated O-specific polysaccharide (O-antigen) was obtained by mild acid degradation of Proteus vulgaris O12 lipopolysaccharide and studied by sugar and methylation analyses, 1H-, 13C- and 31P-NMR spectroscopy, including two-dimensional COSY, TOCSY, NOESY, H-detected 1H, 13C and 1H, 31P heteronuclear multiple-quantum coherence experiments. It was found that the polysaccharide consists of pentasaccharide repeating units connected via a glycerol phosphate group, and has the following structure: where FucNAc is 2-acetamido-2,6-dideoxygalactose and the degree of O-acetylation at position 4 of GalNAc is approximately 25%. Immunochemical studies with P. vulgaris O12 O-antiserum suggested that the lipopolysaccharide studied shares common epitopes with the lipopolysaccharide core of P. vulgaris O8 and with the O-antigens of P. penneri strains 8 and 63.

A neutral O-specific polysaccharide (O-antigen) was isolated from the lipopolysaccharide (LPS) of the bacterium Proteus penneri 71. On the basis of sugar analysis and 1H- and 13C-NMR spectroscopic studies, including two-dimensional COSY, 13C,1H heteronuclear COSY and ROESY, the following structure of the trisaccharide repeating unit of the polysaccharide was established: -->3)-beta-D-GlcpNAc-(1-->4)-beta-D-GlcpNAc-(1-->3)-alpha-D-Galp-(1-- > The polysaccharide has the same carbohydrate backbone as the O-specific polysaccharide of P. penneri 19 and both are similar to that of P. penneri 62 studied by us previously. A cross-reactivity of anti-P. penneri 71, 19 and 62 O-antisera with 11 P. penneri strains was revealed and substantiated at the level of the O-antigen structures. These strains could be divided into three subgroups within a new proposed Proteus O64 serogroup containing P. penneri strains only.

An acidic O-specific polysaccharide was obtained by mild acid degradation of the Proteus penneri 8 lipopolysaccharide and found to contain D-glucose, D-galacturonic acid, 2-acetamido-2-deoxy-D-glucose, 2-acetamido-2-deoxy-D-galactose, 2-acetamido-2,6-dideoxy-L-galactose (L-FucNAc) and 2-aminoethyl phosphate (PEtn) in the ratios 2 : 1 : 1 : 1 : 1 : 1. 1H and 13C NMR spectroscopy was applied to the intact and dephosphorylated polysaccharides, and the following structure of the hexasaccharide repeating unit was established: The O-specific polysaccharide has a unique structure, and, accordingly, we propose for P. penneri 8 a new Proteus O67 serogroup, in which this strain is at present the single representative. The nature of epitopes on LPS of P. penneri 34, P. mirabilis O16, P. mirabilis O23 and P. vulgaris O22, which cross-react with O-antiserum against P. penneri 8, is discussed.

Mucosal immunity was established in the rat prostate by stimulating the common mucosal system through serosal exposure of formalin-killed Escherichia coli. Immunized but not sham-immunized rats developed bacterial specific IgG and IgA in prostatic fluid, and IgA in urine. Immunized (n = 21) and sham-immunized control rats (n = 30) were challenged by transurethral injection of E. coli into the prostate ducts. Mortality, gross and microscopic pathology, tissue bacterial counts, bacterial associated immunoglobulins, and antibody titers in serum and urine were assessed at 7 days following the challenge. Increased E. coli specific immunoglobulin titers were seen in immunized rats, and E. coli, but not Proteus, found in the prostates of immunized animals were coated with IgG and IgA. Immunization protected against toxaemia and septicemia, seen as a rare complication of acute prostatitis, but did not protect against acute prostatitis, nor alter the degree of tissue damage seen in the rat model.

Approximately 4% of peripheral blood T cells in humans express a T cell receptor with markedly restricted germline gene segment usage (V gamma 2 V delta 2). Remarkably, these T cells expand 2- to 10-fold (8%-60% of all circulating T cells) during many microbial infections. We show here that these T cells recognize a family of naturally occurring primary alkylamines in a TCR-dependent manner. These antigenic alkylamines are secreted to millimolar concentrations in bacterial supernatants and are found in certain edible plants. Given the large numbers of memory V gamma 2 V delta 2 T cells in adult humans, recognition of alkylamine antigens offers the immune system a response of the magnitude of major superantigens for alpha beta T cells and may bridge the gap between innate and adaptive immunity.

The acidic O-specific polysaccharide of Proteus vulgaris O22 was studied using 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, NOESY, and H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC) experiments, and the following structure for the branched pentasaccharide repeating unit was established: [sequence: see text] where Qui3NAc is 3-acetamido-3,6-dideoxyglucose, O-acetylation of QuiNAc at position 4 is stoichiometric and at position 2 nonstoichiometric. Serological relationships of P. vulgaris O22 with some other Proteus strains were substantiated on the level of the O-antigen structures.

The O-specific polysaccharide chain (O-antigen) of Proteus penneri strain 22 lipopolysaccharide was studied using chemical methods, including partial acid hydrolysis and Smith degradation, as well as one- and two-dimensional 1H and 13C NMR spectroscopy. The following structure of the pentasaccharide repeating unit was established: [sequence: see text] The O-specific polysaccharide contains a GalNAc residue in the furanose form which has not been hitherto found in bacterial polysaccharides. The O-antigen studied is serologically and structurally unique among Proteus strains and, therefore, a new Proteus serogroup O63 is proposed for P. penneri strain 22.

A neutral O-specific polysaccharide obtained from the lipopolysaccharide of Proteus penneri strain 26 was studied using sugar analysis and 1H and 13C NMR spectroscopy, including two-dimensional NMR techniques. The following structure of the trisaccharide repeating unit was established: -->6)-alpha-D-GlcpNAc-(1-->3)-alpha-L-QuipNAc-(1-->3)-alpha-D-Glcp NAc-(1--> where L-QuiNAc is 2-acetamido-2,6-dideoxy-L-glucose (N-acetyl-L-quinovosamine). Cross-reactivity of the Proteus penneri 26 anti-O serum with other strains of P. penneri isolated in Poland and USA and one strain of P. vulgaris is discussed.

O-specific polysaccharide of Proteus penneri strain 41 was studied using 1H- and 13C-NMR spectroscopy, including two-dimensional COSY, heteronuclear 13C,1H-correlation (HETCOR) and one-dimensional NOE spectroscopy, and the following structure of a non-stoichiometrically O-acetylated hexasaccharide repeating unit was established:[structure: see text] where RGlcNAc is 2-acetamido-4-O-[(S)-1-carboxyethyl]-2-deoxyglucose. Cross-reactivity of anti-P. penneri 41 O-serum with other P. penneri strains is discussed, and a new, separate O62 serogroup is proposed which is the next Proteus O-serogroup containing P. penneri strains only.

A total of 257 Proteus strains isolated from urinary tract infection, blood, wound and faeces were studied. Of the strains tested 31 (12 percent) were serum sensitive, 182 (71 percent) were serum resistant and the remaining 44 (17 percent) showed intermediate sensitivity to the pooled normal human serum (PNHS). Strains isolated from adult urines and blood cultures were significantly more sensitive than strains of faecal origin (p < 0.01). No significant difference was seen between strains from faeces and wounds.

On the basis of sugar analysis and 1H- and 13C-NMR spectroscopy, it was shown that the O-specific polysaccharide of Proteus penneri strain 15 has a trisaccharide repeating unit, including an acetal-linked pyruvic acid residue, and is structurally identical to the capsular polysaccharide of Proteus vulgaris strain ATCC 49990. Serological studies supported this conclusion and demonstrated the presence in the homological antiserum of both anti-core and anti-O chain antibodies reacting with a lipopolysaccharide (LPS) epitope containing N-acetylglucosamine and galactose residues.

Objective: To determine whether patients with ankylosing spondylitis (AS) and patients with rheumatoid arthritis (RA) from Japan have antibodies to Klebsiella pneumoniae and Proteus mirabilis and to assess whether such antibodies are activated against peptides sharing sequences with HLA-B27., Methods: Serum samples from 152 Japanese patients, 52 with AS, 50 with RA, and 50 healthy controls, were tested against 3 bacteria (K. pneumoniae, P. mirabilis, and Escherichia coli) and 3 synthetic peptides (HLA-B27, pullulanase-D, and scrambled pullulanase-D control peptide) by ELISA under coded conditions. Samples were tested for elevations in IgG, IgA, and IgM antibody classes in patients with active AS or RA, in patients with RA with probable disease, and in patients with inactive AS. Disease activity was determined by an elevated serum C-reactive protein (> 10 mg/l) level and elevated erythrocyte sedimentation rate (> 20 mm/h)., Results: Patients with active AS showed specific elevations in serum IgA antibody levels against K. pneumoniae compared to patients with RA and controls (p < 0.001). No such elevation was seen in the IgG and IgM antibody classes. Patients with inactive AS showed no elevation in any class of antibody against K. pneumoniae compared to controls or patients with RA. Patients with active or probably active RA showed significant elevations in IgG antibody levels against P. mirabilis compared to AS and controls (p < 0.001). Patients with AS (active or inactive), RA (active or probably active), and controls showed no elevations in any antibody class to E. coli. Both active and inactive AS patients had specific autoantibodies against HLA-B27 peptide compared to patients with RA and controls (active AS: IgG, IgA, IgM, p < 0.001; inactive AS: IgG and IgA, p < 0.001). Patients with active AS had IgG and IgA antibodies against pullulanase-D peptide, which contains a sequence that cross reacts with HLA-B27 compared to controls (p < 0.001)., Conclusion: These results provide the first evidence of AS and RA patients in Japan having specific elevations of antibody to K. pneumoniae and P. mirabilis, respectively. This suggests that K. pneumoniae in AS and P. mirabilis in RA may play a role in triggering and/or exacerbating these diseases.

Immunological investigation (1-2 level tests with monoclonal antibodies) of 246 OMPC patients has discovered a relationship between the degree of immunological disorder, blood group and microflora. Patients carrying AII and ABIV, Pseudomonas aeruginosa, Staphylococcus aureus, Proteus were more immunologically compromised. These findings may help in the choice of profile immunocorrector.

An acidic O-specific polysaccharide isolated from Proteus penneri 52 contains D-galacturonic acid, 2-acetamido-2-deoxy-D-glucose, and 2-acetamido-2-deoxy-D-galactose in the ratio 1:2:2. On the basis of sugar analysis and NMR spectroscopy, which included one-dimensional TOCSY, two-dimensional COSY, heteronuclear 13C, 1H-COSY, and rotating-frame NOE spectroscopy, the following structure of the pentasaccharide repeating unit of the O-specific polysaccharide was established: [sequence: see text] Cross-reactivity of the anti-P-penneri 52 O-serum with other strains of P. penneri isolated in Germany. Poland, USA, and Canada is discussed and a new Proteus serogroup is proposed.

Objectives: To investigate the role of potential immunoselected phages displaying random peptides in addition to possible antigen leads in rheumatoid arthritis (RA) by assaying the levels of synovial fluid (SF) and serum antibodies to synthetic peptides., Methods: Serum and SF antibodies from patients and controls were measured using an enzyme linked immunosorbent assay (ELISA)., Results: Sera and SF from RA patients reacted significantly more strongly to a 12 amino acid peptide, EFHELGDIAIAA, that shares a significant homology with collagen type IX, than did SF and sera from control groups (p < 0.0209 and p < 0.0115, respectively). In addition, the humoral responses to a 15 amino acid peptide, GGYGDGGAHGGGYGG, derived from the glycine-rich cell wall protein (GRP) 1.8, and to a 16 amino acid synthetic peptide, LGSISESRRALQDSQR, derived from the Proteus haemolysin protein were significantly stronger in RA patients compared with healthy individuals (p < 0.0001 and p < 0.0011, respectively)., Conclusion: Our data indicate that peptide phage libraries can be used as tools for the identification of the (auto)antigen leads that may be responsible for the initiation, perpetuation, or both, of the immune response in patients with RA.

The bacteria Proteus, Serratia, Escherichia and Pseudomonas possess sequences resembling the rheumatoid arthritis susceptibility sequence EQRRAA, but antibodies were elevated only to Proteus in 66 RA patients (P<0.001) when compared to 61 active ankylosing spondylitis patients and 60 controls.

The treatment of 17 patients having chronic obstructive bronchitis with a combined vaccine resulted in a rise in the number of T lymphocytes and T suppressors, as well as a simultaneous increase in their functional activity, in these patients. Moreover, an increase in the ingestive activity of neutrophils and in the concentration of immune complexes was registered. Treatment with the vaccine containing Staphylococcus aureus, Klebsiella pneumoniae, Proteus and Escherichia coli antigens facilitated an increase in the titers of antibodies to most antigens contained in the vaccine and in the resistance of the body to infection. Vaccinal therapy made it possible to achieve a significant prolongation of the period of remission.

The chemical structure of the O-specific polysaccharide chain of Proteus penneri 62 lipopolysaccharide (LPS) containing N-acetylisomuramic acid was established using acid hydrolysis, solvolysis with anhydrous hydrogen fluoride and 1H and 13C NMR spectroscopy. Cross reactivity of the anti-O-serum P. penneri 62 with a number of other strains of the same species isolated in the USA, Canada, Germany and Poland is discussed.

The reactivity of anti-Rickettsiae human antibodies with Proteus OX cells is used as a presumptive rickettsial diseases diagnostic test Weil-Felix reaction. In presented studies we compare the reactivity of human anti-Rickettsiae and rabbit anti-Proteus antibodies with series of Proteus OX2, OX19 and OXK lipopolysaccharides (LPS). Polyclonal rabbit anti-OX2, anti-OX19 and anti-OXK sera reacted only with the homologous LPS--OX2, OX19 and OXK, respectively. The antibodies of Japanese spotted fever patients were less specific and reacted with OX2 as well as OX19 LPS. The antibodies of scrub typhus patients recognized Proteus OXK LPS, only. The serological reactions of O-antigen of P. mirabilis S1959 indicated that this, previously serologically not classified, strain may belong to the OXK group. Bacteria, used in the studies, came from the American, Japanese and European strain collections. The series of OX2, OX19 and OXK LPS, isolated from these Proteus strains, presented pattern of electrophoretic mobility and serological reactivities specific for each of OX-group types.

We wished to resolve a paradox of how the response to the phosphocholine (PC) determinant of Proteus morganii could be initiated from a precursor B cell whose receptor, in unmutated form as Ab, appears to be unable to bind Ag. Unmutated VH and unmutated VL constructs were co-transfected into the B cell lymphoma M12.4 to study stimulation via membrane lg (mlg). The same VH construct was expressed in an L+, H- hybridoma line to characterize Ab binding. The unmutated Ab showed no detectable binding in ELISA to the PC-containing Ag from P. morganii PC(PM). By contrast, the unmutated mlg mediated mobilization of calcium in response to the PC(PM) Ag. Single-positive B cell lines of mlgM, mlgD double-positive lines were all capable of responding. The degree of signaling depended greatly on high receptor number, and only a fraction of cells in the population responded. Inhibition of the PC(PM)-induced calcium response by free PC indicated that the response was Ag-specific. A transfectant B cell line expressing moderate levels of a high affinity, mutated mlgM readily responded to PC(PM). These observations indicate that the unmutated lg as a receptor is capable of interacting with PC(PM) and suggest that the immune response to PC(PM) could originate from the precursor B cell expressing the unmutated mlg. The role of mlgD vs mlgM is discussed in terms of the requirement for high receptor number in the signaling process.

Biochemical and antigenical characteristics of tannin-protein complex degrading enterobacteria (T-PCDE) isolated from Koalas, Phascolarctos cinereus, were investigated. T-PCDE had a specific profile of characteristics, and T-PCDE was distinguished from those of 12 type strains of Enterobacteriaceae used.

In the approbation of the enzyme-linked immunosorbent assay (ELISA) system on the basis of F(ab)2 fragments of antistaphylococcal antibodies on 307 cultures of the representatives of the genera Staphylococcus, Pseudomonas and Proteus high sensitivity, specificity and effectiveness of ELISA for the quantitative and qualitative evaluation of the alpha-hemolytic activity of S.aureus were established. The ELISA system has made it possible to additionally detect alpha-hemolysin in 62% of S.aureus strains classified with as nontoxigenic strains using hemolysis test in Petri dishes. The sensitivity limit of this method is 0.0005 binding units or 1.0 ng in terms of protein content. The use of the ELISA system may be recommended for the study of the toxigenic properties of staphylococci.

Peripheral blood leukocytes of intact dogs, when transplanted to sublethally irradiated mice, have been found to suppress endogenous colony formation. The inactivation effect in hemopoietic stem cells of mice is enhanced with an increase in the number of effector cells in the graft. Radiation exposure results in failure of the effector function of leukocytic cells at the peak of acute radiation sickness. Nine months after irradiation, the inactivating capacity of dog leukocytes is restored and exceeds that of the control. Administered to dogs 9 months after irradiation, proteus vaccine stimulated the effector function of leukocytes at days 7 and 28 and suppressed it at days 14 and 62 post-administration.

Sera from 4 out of 19 patients with the Japanese spotted fever were negative to OX2 antigen of Weil-Felix (WF) test. These WF test negative sera were analyzed by ELISA and immunoblot used whole cells and lipopolysaccharides (LPS) of rickettsiae and Proteus strains as antigens. These acute-phase sera have already possessed the IgG antibodies against LPS of Proteus OX2 strain, whereas IgM antibodies in these acute- and convalescent-phase sera did not react with this LPS. On the other hand, the reactivity of IgM antibodies of the convalescent-phase sera in the 2 patients with LPS of Proteus OX19 strain increased as compared with that of the acute-phase sera by ELISA, and these IgM antibodies also showed the reactivity with bands of OX19-LPS in the immunoblot. On the basis of these results, it is interpreted that the WF test negative sera from patients with Japanese spotted fever are due to the presence of IgG antibodies against OX2-LPS in the sera.