Commensal intestinal bacterial strains trigger ankylosing enthesopathy of the ankle in inbred B10.BR (H-2(k)) male mice.

Joint disease ankylosing enthesopathy (ANKENT) naturally occurs in inbred mice with C57Bl/10 genetic background. ANKENT has many parallels to human ankylosing spondylitis (AS) and represents an animal model for AS. Environmental conditions (i.e., microbial load of the organism) are among the risk factors for ANKENT, similar to AS. The role of microflora in the development of ANKENT was investigated. ANKENT was tested in four experimental groups of germ-free mice associated with different numbers of various intestinal microbes and three control groups: germ-free, specific pathogen-free, and conventional (CV) mice. Mice were colonized either with anaerobic bacteria isolated from the intestine of a CV mouse or with bacterial strains obtained from the collection of microorganisms. Microbes were characterized and checked by microbiological cultivation methods and with the use of polymerase chain reaction amplification and rDNA sequence analysis. Joint disease developed in GF mice colonized with a mixture containing Bacteroides spp. and Enterococcus sp., and/or Veillonella sp. and Staphylococcus sp. No ANKENT appeared in males colonized with probiotic bacterium Lactobacillus sp. In control groups ANKENT occurred in SPF and CV animals; the GF animals remained healthy. The results confirmed that the germ-free conditions protect from joint inflammation, and thus microbes are necessary for ANKENT development. In colonized mice the ANKENT was triggered by luminal anaerobic bacteria, which are common components of intestinal microflora.

Psychiatrist's adherence: a new factor in relapse prevention of schizophrenia. A randomized controlled study on relapse control through telemedicine system.

ACCESSIBLE SUMMARY: Exposure to psychotic states has detrimental effects on the long-term outcome of schizophrenia and brain integrity. Therefore, improving relapse prevention is a key component of long-term management of schizophrenia. Previous studies using continuous monitoring of an individual's early signs of relapse and adopting preventative pharmacological interventions, when early signs are detected, showed promising clinical results in terms of relapse risk reduction. This 18-month multi-centre parallel randomized controlled, open label, trial with telemedicine relapse prevention programme ITAREPS failed to show superiority of maintenance plus prodrome-based targeted medication strategy over treatment as usual. The study, marked by low investigator's adherence, confirmed that absence of pharmacological intervention at early stage of prodrome, critically influenced the risk of relapse. This and previous randomized controlled trials with telemedicine programme ITAREPS suggested that substantial improvement in relapse prevention in schizophrenia is likely to be unattainable under current clinical settings. Future preventive strategies in schizophrenia would require rapid pharmacological intervention upon occurrence of subclinical prodromal symptoms that are undetectable under conventional outpatient practice. Studies with ITAREPS suggested that integration of telemedicine relapse prevention systems and visiting nurse service might together represent practical solution capable to address those requirements. ABSTRACT: The Information Technology Aided Relapse Prevention Programme in Schizophrenia (ITAREPS) presents a telemedicine solution for weekly monitoring and management of schizophrenia. This study aims to evaluate the effectiveness of the programme in reducing the number of hospitalizations during the 18-month multi-centre parallel randomized controlled, open label, trial. Outpatients with schizophrenia or schizoaffective disorder were randomized to the active (n = 74) or control group (n = 72). In the active arm, investigators increased the antipsychotic dose upon occurrence of prodrome announced by the system. Intention-to-treat analysis showed no between-group difference in the hospitalization-free survival rate [Kaplan-Meier method; hazard ratio (HR) = 1.21, 95% confidence interval (CI): 0.56-2.61, P = 0.6). In a post hoc multivariate Cox proportional hazards model, out of 13 potential predictors, only ITAREPS-related variables (number of alerts without pharmacological intervention/HR = 1.38, P = 0.042/ and patient non-adherence with ITAREPS /HR = 1.08, P = 0.009/) increased the risk of hospitalization. In this trial ITAREPS was not effective. The results in context with previous ITAREPS studies suggest non-adherence of both psychiatrists and patients as the main reasons for the failure of this preventive strategy. Tertiary prevention in schizophrenia have to be regarded a major challenge, warranting the need for implementation of strategies with more active participation of both patient and treating psychiatrist.

Germ-free mice do not develop ankylosing enthesopathy, a spontaneous joint disease.

Ankylosing enthesopathy (ANKENT) is a naturally occurring joint disease in mice with numerous parallels to human ankylosing spondylitis (AS). Similarities between AS and ANKENT include not only affected tissue (joint entheses) but also association of the disease with genetic background, including MHC genes, gender, and age. Young males with the C57Bl/10 background have been described to suffer from ANKENT and, among H-2 congenic strains, high frequency of afflicted joints has been recorded in B10.BR (H-2(k)) males. Interestingly, the incidence of ANKENT is higher in conventional (CV) males that in their specific-pathogen-free (SPF) counterparts. The latter finding suggests that microbes could play a role as an ANKENT-triggering agent. To further examine this hypothesis we have established a germ-free (GF) colony of B10.BR mice and observed ANKENT incidence in both GF males and their conventionalized (ex-GF) male littermates; 20% of ex-GF males developed ANKENT before 1 year of age. In contrast, no joint disease was observed under GF conditions (p < 0.0001). Our results show that live microflora is required in ANKENT pathogenesis.

HLA-B27 as a relative risk factor in ankylosing enthesopathy in transgenic mice.

HLA-B27 is a risk factor for several human diseases through a mechanism that is not yet understood. This article describes a naturally occurring joint disease in laboratory mice, ANKENT. ANKENT begins with mild inflammation and culminates in irreversible stiffening of the ankle and/or tarsal joints in one or both hind paws. The macroscopic and histologic features of ANKENT, its relationship to age, gender, and environment, and some immunologic aspects are considered. With respect to genetics, it is demonstrated that an HLA-B27 transgene is a relative risk factor for ANKENT. Its impact depends on the H-2 haplotype, reaching a relative risk value of 9.4 for C57Bl/10, H-2b males (p < 0.025). Several features of ANKENT are reminiscent of human AS: joint pathology, age and gender distribution, the presence of non-MHC as well as MHC risk factors (including HLA-B27), and the suspicion that environmental factors are involved.

Strong association of HLA-B27 heavy chain with beta 2-microglobulin.

Monoclonal antibody TG1 recognizes specifically antigens HLA-B27, B7, B22 and B17 on cell surface in cytotoxicity and cytofluorometry tests. When cell detergent extracts were subjected to SDS PAGE under mild conditions (no heating and no reduction of the sample) followed by Western blotting, TG1 detected exclusively a complex of B27 heavy chains with beta(2)-microglobulin (as a 50 kDa band) whereas the other B-locus antigens (B7, B22, B17) were detected as free 43 kDa heavy chains under the same conditions. When the samples were boiled prior to SDS PAGE, TG1 detected again the 43 kDa free heavy chains of B7, B22 and B17 but no zone corresponding to B27 could be detected indicating that the epitope in free B27 chains is more sensitive to denaturation by SDS. Thus, our main finding is that the interaction of HLA-B27 heavy chain with beta(2)-microglobulin appears to be stronger than that of the other HLA-B chains. The resistance of the HLA-B27/beta(2)-microglobulin complex to the SDS dissociation is strikingly similar to the behavior of MHC class II molecules under similar conditions. Thus, it may be speculated that HLA-B27 complexes can be also more stable than other MHC class I molecules under more physiological dissociative conditions (e.g. in endosomal compartments). This feature might potentially influence antigen presentation by HLA-B27 and contribute to the well known disease linkage of HLA-B27.

The cell growth inhibitory and antiviral effects of interferon in cloned transformed mouse cells.

Eight clones and two subclones of SV40- and seven clones and one subclone of 20-methylcholanthrene-transformed C3H mouse embryonic fibroblasts were compared in tests for sensitivity to the antiviral and cell-growth inhibitory activities of a partially purified mouse L-cell interferon. While the sensitivity of clones and subclones to the antiviral activity of interferon was comparable to that of parent lines, the cell-growth inhibitory activity of interferon in the SV40 clones showed more than 100-fold variation and the methylcholanthrene-transformed cells could be divided into two groups in this respect. No correlation of sensitivity to the cell-growth inhibitory effect of interferon with the chromosome number, interferon-producing capacity or tumorigenicity of the clones could be detected. However, the cells of the interferon-sensitive clones No. 36 of the methylcholanthrene-transformed line were destroyed by macrophages at higher percentage binding of 125I-labeled soybean lectin. These results suggest that (1) the cell-growth inhibitory effect of interferon might be mediated by a specific type of receptors, and (2) N-acetyl-galactosamine present on the surface of interferon-resistant cells in a higher concentration than on interferon-sensitive cells hinders the recognition of cells both by macrophages and by interferon.

A new H-2 congenic strain B10.ABQ/Ph carrying a recombinant H-2 haplotype derived from balanced lethal stock Ttf/t12.

A new H-2 congenic strain, B10.ABQ/Ph, has been established. It carries the recombinant H-2 haplotype derived from balanced lethal stock Ttf/t12. The recombinant H-2 haplotype was designated bq5 and possesses the following alleles at individual loci of the H-2 complex: Kb A beta b A alpha b E beta b or q E alpha b or q Sb or q Dq Lq.

H-2 influence on ankylosing enthesopathy of the ankle (ANKENT).

Joint disease, ankylosing enthesopathy (ANKENT) of the ankle, occurs naturally in mice. ANKENT depends on the genetical constitution, sex, and age, and its frequency varies among inbred strains of normal mice. We found the highest frequency of ANKENT in B10.BR (H-2k) males. The H-2k haplotype appears to be a relative risk factor, which increases the susceptibility to ANKENT. The aetiology of ANKENT is unknown but an involvement of microbial agents in the environment is supposed.

Monoclonal antibody to human sperm acrosomal protein.

A new monoclonal antibody designated Hs-14 was generated after immunization of BALB/c mice with the acid extract of human sperm. In indirect immunofluorescence Hs-14 mAb binds to the acrosome of permeabilized sperm cells and consequently recognizes some intra-acrosomal protein. Western blotting analysis revealed that under non-reducing conditions the Hs-14 mAb detects a protein with a molecular mass of 220 kDa. Under reducing conditions the Hs-14 recognizes several peptide bands within the range from 55 kDa to 110 kDa. Beside human sperm the antibody positively reacts also with sperm of some other mammalian species. Using Hs-14 mAb it is possible to evaluate the acrosomal integrity of spermatozoa and to reveal sperm pathology.

Recessive lethal t haplotypes increase the frequency of the partial trisomy of chromosome 17 (including the T-t complex) among offspring of T(16;17)43H female mice.

The male-sterile reciprocal autosomal translocation T(16;17)43H displays a high frequency of adjacent-2 disjunction in meiosis of translocation heterozygotes. One haploid product of this abnormal chromosome segregation leads to viable partial trisomy of chromosome 17 after fertilization. The frequency of trisomics increases in the progeny of T43H/+ females when the wild-type allelic form of T-t complex is substituted for t12, tw32 or t6 recessive lethal haplotypes. Indirect evidence supports the idea that the suppression of crossing-over by t haplotypes is linked with the increase in adjacent-2 disjunction and subsequent Ts43H trisomy. The possible use of Ts43H trisomy for genetic dissection of the T-t complex is briefly discussed.

Genetic conditions for the release of a lymphokine "lymph node activating factor".

The release of a lymph node activating factor is controlled by the genetic difference in the I region of the H-2 complex. Incompatibility at IA and IE loci is of decisive importance.

Preparation and characterization of a monoclonal antibody against boar acrosin.

BALB/c mice were immunized with the crude fraction of boar acrosin. Immune spleen cells were fused with myeloma cells SP2/0-Ag14. One of the 6 hybridomas produced was cloned and characterized by ELISA and SDS-PAGE. Possible uses of the monoclonal antibody against acrosin for immunological detection of the amount of acrosin liberated after manipulations with spermatozoa and for selection of undamaged spermatozoa for insemination are discussed.

Interferon-induced changes in the expression and topography of H-2 antigens and receptors for SRBC in tumour cells.

The expression of H-2 antigens has been investigated in methylcholanthrene-transformed C3H embryonic fibroblasts of clones 36 and 57. Private specificities H-2.23 and H-2.32, typical of the H-2k haplotype, were present on cells of the two clones but differed considerably in their expression. After interferon treatment, the expression of H-2 antigens increased in both clones, though more markedly in clone 57. Interferon treatment also affected the topography of H-2 antigenic determinants on the cell membrane. The blocking index for antigenic determinants coded by a single region of the H-2 complex increased in both clones. Between antigenic determinants controlled by two regions of H-2 the blocking index remained unaltered in clone 36 but decreased significantly in clone 57. Rosette formation with SRBC was consistent with the results of the blocking test between H-2K and H-2D antigenic determinants; rosette formation decreased slightly in clone 36 but increased in clone 57 after interferon treatment. All the results suggest that alterations in expression of H-2 antigens following interferon treatment are due to the rearrangement of the cell membrane rather than to the virtual increase in the number of H-2 antigenic determinants. Cells of clone 57 exhibiting more pronounced changes in the topography of H-2 antigens were more resistant to the inhibitory effect of interferon on cell growth.

Alloreactive cytotoxic anti-H-2 antibodies in sera of the syngeneically immunized C57BL/10ScSnPh, BALB/cJPh, and B10.A/SnPh mice.

Mice of the inbred strains B10.A (H-2a), B10 (H-2b), and BALB/c (H-2d) were immunized with the syngeneic cells, and the sera obtained from individual animals on day 10 after the 8th immunizing injection were assayed for the presence of alloreactive cytotoxic antibodies. Anti-H-2Kk antibodies were present in the sera from 50% of the syngeneically immunized BALB/c mice. The syngeneic-immune B10 sera contained weak anti-H-2Dd antibodies in 14% of the immunized animals. B10.A mice produced no antibodies after syngeneic immunization.

Monoclonal antibodies to intra-acrosomal proteins inhibit gamete binding in vitro.

The sperm-zona pellucida-binding assay in vitro was used as a functional test for zona pellucida-binding ability of boar spermatozoa after co-incubation with monoclonal antibodies against intra-acrosomal proteins. The effect of monoclonal antibodies ACR.2 against boar acrosin (55, 53, 45 and 38 kDa), and Hs-8 against boar intra-acrosomal protein (230, 110, 88, 60, 48 kDa) on boar spermatozoa-porcine oocyte binding was examined. The sperm-zona pellucida-binding was reduced when medium was supplemented with monoclonal antibodies during sperm-oocyte co-incubation, but not when capacitated spermatozoa were pretreated with monoclonal antibodies before incubation with oocytes. Our results show that the monoclonal antibodies (ACR.2, Hs-8) against intra-acrosomal proteins reduce the secondary sperm-zona pellucida-binding with statistically significant difference. This suggests the role of these proteins in the early phases of fertilization.

Irradiation of genetically modified plasmacytoma vaccines results in upregulation of CD80 molecule expression, IL-2 production and higher therapeutic efficacy of the vaccines.

It has been found previously that irradiated, IL-2 gene-modified plasmacytoma (X63-m-IL-2) vaccines are more efficient in the therapy of the parental (X63-Ag8.653) plasmacytoma than live plasmacytoma vaccines. In this communication, we have demonstrated that irradiation of murine IL-2-producing plasmacytoma vaccines resulted in upregulation of CD80 molecule expression and IL-2 production. The expression of MHC class I antigens was not altered. The upregulation of the CD80 membrane molecule expression in X63-m-IL-2 cells was higher after irradiation with 150 Gy than after irradiation with 50 Gy. Comparable upregulation of the CD80 molecule expression has also been demonstrated after irradiation of the parental murine X63-Ag8.653 plasmacytoma cells. The results indicate that upregulation of the CD80 molecule expression and enhanced IL-2 production in irradiated X63-m-IL-2 cells was responsible for the higher therapeutic effectiveness of the irradiated plasmacytoma vaccine.

Experimental colitis does not increase the prevalence of ANKENT, a spontaneous joint disease in mice.

A possible relationship between intestinal inflammation and joint disease development was investigated. Clinical symptoms of colitis--diarrhea and rectal bleeding--were confirmed by findings of inflammatory processes in the colon in dextran sodium sulfate-treated mice and joint ankylosing enthesopathy (ANKENT) developed in 12.8 % mice with chronic colitis and 13.6 % mice in the control group. Consequently no significant difference in ANKENT frequency was found between mice with and without chronic colitis and the occurrence of ANKENT in both groups was typical for conventional conditions. ANKENT cannot be triggered solely a generalized inflammatory process in the gut.

[Use of human sperm protein monoclonal antibodies in the diagnosis of sperm pathology and selection of a suitable assisted reproduction method for fertilization]. / Vyuzití monoklonálních protilátek proti proteinum lidských spermií k diagnostice patologie spermií a výberu vhodné metody fertilizace v asistované reprodukci.

OBJECTIVE: Use of monoclonal antibodies against sperm proteins in human medicine. DESIGN: Experimental and clinical studies. SETTING: Dep. Biology and Biochemistry of Fertilization, Institute of Molecular Genetics, Prague, Laboratory IVF, Iscare IVF, Prague, Dep. of Immunobiology, Institute for the Care of Mother and Child, Prague. METHODS: Monoclonal antibodies against human sperm intra-acrosomal and cell surface proteins were used for quantitative analysis of these proteins by the immunofluorescence test in samples of human sperm of good and poor qualities. RESULTS: The detection of intra-acrosomal proteins was decreased and, on the other hand, detection of surface proteins was the same or higher in pathological spermatozoa. CONCLUSIONS: Monoclonal antibodies can be used for diagnostics of sperm pathology (quantitative detection of proteins) and for evaluation of the physiological state of sperm cells (state of acrosome before or after acrosome reaction). Finally, monoclonal antibodies could be useful for selection of a suitable method of fertilization (IUI, standard IVF, ICSI) in the laboratories of assisted reproduction.

[Intraspecies identification of cultured murine cells by using H-2 antigen typing]. / Vnutrividovaia identifikatsiia kul'tiviruemykh kletok myshi s pomoshch'iu tipirovaniia H-2-antigenov.

17 mouse cell lines have been screened with specific sera against H-2 antigens. All the cell lines tested expressed H-2 antigens characteristic of the donor haplotype. The data obtained indicate that H-2 typing of cultured mouse cells can be used as an approach to control their intraspecies diversity.

[Recombinant haplotype H-2 in mice of the congenic resistant strain B10.D1(R108)/Y]. / Izuchenie rekombinantnogo gaplotipa H-2 myshei kongenno-rezistentnoi linii B10.D1(R108)/Y.

Recombinant H-2 haplotype of mouse strain B10.D1(R108)/Y (symbol R108) obtained in experiments with skin grafting in the course of developing the CR B10.D1/Y strain (strain DBA/LacY--the donor of H-2q) was studied. Strains with recombinant H-2 haplotypes a, h2, g1, i3, i5, i7, m, y1 were used. Alleles of different H-2 (K, I, D) regions were determined according to the presence or absence of genetic complementation in the F1 test with skin grafts. R108 recombinant was studied by serological methods with panel of anti-H-2 sera. Anti-H-2Kb (H-2.33) and anti-H-2Dq (H-2.30) monospecific antisera were used in microcytotoxicity test and in absorption experiments in vitro. It was concluded that crossing over between H-2b and H-2q chromosomes, which led to formation of recombinant H-2 haplotype of R108 mice, occurred at I region, between IA and IC subregions. The H-2 complex of R108 line has KbIAbIJ?IE?ICqSqDq alleles. bq1 symbol was proposed for the H-2 haplotype of B10.D1(R108)/Y strain.