Carbon monoxide inhibits T cell activation in target organs during systemic lupus erythematosus.

Systemic lupus erythematosus is characterized by the presence of circulating anti-nuclear antibodies (ANA) and systemic damage that includes nephritis, haematological manifestations and pulmonary compromise, among others. Although major progress has been made in elucidating the molecular mechanisms responsible for autoimmunity, current therapies for lupus have not improved considerably. Because the exposure of carbon monoxide (CO) has been shown to display beneficial immunoregulatory properties in different immune-mediated diseases, we investigated whether CO therapy improves lupus-related kidney injury in lupus mice. MRL-Fas(lpr) lupus mice were exposed to CO and disease progression was evaluated. ANA, leucocyte-infiltrating populations in spleen, kidney and lung and kidney lesions, were measured. CO therapy significantly decreased the frequency of activated B220(+) CD4(-) CD8(-) T cells in kidneys and lungs, as well as serum levels of ANA. Furthermore, we observed that CO therapy reduced kidney injury by decreasing proliferative glomerular damage and immune complexes deposition, decreased proinflammatory cytokine production and finally delayed the impairment of kidney function. CO exposure ameliorates kidney and lung leucocyte infiltration and delays kidney disease in MRL-Fas(lpr) lupus mice. Our data support the notion that CO could be explored as a potential new therapy for lupus nephritis.

Chemokine response induced by Chlamydia trachomatis in prostate derived CD45+ and CD45- cells.

The role of innate cells and their receptors within the male genital tract remains poorly understood. Much less is known about the relative contribution of different genital tract cells such as epithelial/stromal cells and resident leucocytes. In this study, we examined innate immune responses to Chlamydia trachomatis by prostate epithelial/stromal cells and prostate resident leucocytes. Murine prostate primary cultures were performed and leucocyte and epithelial/stromal cells were sorted based on surface protein expression of CD45 by magnetism-activated cell sorting or fluorescence-activated cell sorting. Prostate derived CD45- and CD45+ cells were infected with C. trachomatis and chemokine secretion assayed by ELISA. Similar experiments were performed using prostate CD45+ and CD45- cells from myeloid differentiation factor 88 (Myd88(-/-)) mice or toll-like receptor (Tlr2(-/-)) and Tlr4(mutant) double-deficient mice. Moreover, a TLR-signalling pathway array was used to screen changes in different genes involved in TLR-signalling pathways by real-time PCR. Prostate derived CD45- and CD45+ cells responded to chlamydial infection with the production of different chemokines. Both populations expressed genes involved in TLR signalling and required to respond to pathogen-associated molecular patterns and to C. trachomatis infection. Both populations required the adaptor molecule MYD88 to elicit chemokine response against C. trachomatis. TLR2-TLR4 was essential for chemokine production by CD45+ prostate derived cells, but in their absence, CD45- cells still produced significant levels of chemokines. We demonstrate that C. trachomatis is differentially recognised by prostate derived CD45+ and CD45- cells and suggest that diverse strategies are taking place in the local microenvironment of the host in response to the infection.

[The search for Yersinia enterocolitica in vegetables treated by a cold line in school cafeterias in Montpellier]. / Recherche de Yersinia enterocolitica dans des légumes traités par la chaîne de froid des restaurants scolaires de Montpellier.

From 134 samples of raw or cooked vegetables taken in two points of a cold line we isolated 19 strains of Yersinia enterocolitica, 3 of Y. intermedia 3 of Y. kristensenii. None of them belongs to serotypes usually pathogenic for man. The raw vegetables are most contaminated. Y. kristensenii and Y. intermedia were isolated especially from samples of green salad. The used method was a cold enrichment at 4 degrees C followed by a potassium hydroxyde treatment. The activity of 16 antimicrobial drugs was studied by the disk diffusion test at 28 degrees C. All strains were sensitive to aminoglycosides, minocycline, colistine and cotrimoxazole. The results obtained with chloramphenicol and beta-lactam antibiotics vary in terms of species and/or biotypes.

[Study of a brucellosis epidemic in a horticultural school]. / Etude d'une épidémie de brucellose dans une collectivité scolaire horticole.

An epidemiological survey was conducted after the observation of 4 cases of acute brucellosis in an horticultural school. Of a total number of 215 attendants, eventually exposed to a single contamination, 65 cases of brucellosis were thus identified. The source of contamination was probably a practical lesson where the pupils studied a bovine gravid uterus. More stringent regulations are needed in order to avoid further similar epidemics.

Cellular responses of the mouse to the peptidoglycan of a gram-positive bacterium (Streptococcus pyogenes).

The cellular responses and the stimulation of the reticulo-macrophagic system induced in the mouse by a purified bacterial peptidoglycan (PGL) as previously described, were studied by the changes in the peritoneal cytology, the macrophage-migration-inhibition test and the clearance of colloidal carbon. PGL was submitted to chemical and immunochemical characterization and was shown to be substantially free of contamination by polysaccharides, phospholipids, teichoic acid and nucleic acids, but to contain a detectable amount of peptide contaminants; N-acetylglucosamine and the tetrapeptide (with terminal D-alanine) were shown to be the main antigenic determinants. This substance had no action on polymorphonuclear leucocytes but induced an inhibition of the migration of macrophages. This was due to an immunological reaction rather than to direct cytotoxicity, as shown by the negative cytotoxicity tests and the age and life-environment-dependence of the phenomenon. The reticulomacrophagic system was significantly stimulated after primary inoculation, and still more so after a booster. The possible mechanisms of these activities, which are therefore independent from toxic and/or inflammatory responses, are discussed.

[Demonstration of brucella antigens in situ in mouse cells and spleen by electron microscopy]. / Mise en évidence des antigènes brucelliens in situ dans les cellules du ganglion et de la rate de souris par la microscopie électronique

The immunoglobulins of rabbit anti-brucella immunsera were conjugated to horse radish peroxydase and used to revell the bacterial antigens during the experimental infection of mice with Brucella suis. The technique seems to make possible the identification of bacteria that have undergone morphological modifications after phagocytosis.

Kinetics of the uptake of rifampicin and tetracycline into mouse macrophages. In vitro study of the early stages.

The in vitro uptake by mouse peritoneal macrophages, of chlortetracycline (by fluorescence microscopy) and of tetracycline and rifampicin (by scintillation spectrometry of radioactive antibiotics) has been studied over a six hours period, using various concentrations of the antibiotics, close to the therapeutic concentrations. The incidence of the conditions of the assays, especially that of the use of heterologous serum for the cultivation of cells, has been investigated; a medium supplemented with homologous serum at low concentration has been devised with the technique. The uptake of these antibiotics was a three-phases process suggesting the superposition to a passive diffusion of either an active incorporation, or a restriction of the outflow (perhaps associated). This led to a rather high concentration of the antibiotics into cells, although other studies have shown that this concentration is not as active on intracellular bacteria as one could expect from the in vitro sensitivity.

Response of the lymphoid organs of the mouse to the peptidoglycan of a gram-positive bacterium (Streptococcus pyogenes).

The response of the lymphoid organs of the mouse to the injection of a Gram-positive (Streptococcus pyogenes) bacterial peptidoglycan (PGL) has been investigated by several complementary techniques. Special cares were brought to: a) the extraction mode and the characterization of the bacterial fraction, b) the sanitary and microbiological status of the mice, and c) the lack of biologically active contaminants, even at a trace level. In these conditions, the local reaction at the site of inoculation was very moderate, transient and not very distinctive. No toxic reactions were observed, even with doses of several milligrams per mouse. Changes in lymphoid organs, as determined by changes in weight, incorporation of radioactive precursors and detailed histological study revealed that PGL did not induce the typical responses of ordinary antigens. Changes in the weight of organs were not significant and there was no dose-response relationship. In the lymph node, the response was confined to the paracortical area; the lymphocyte activation stood at an early stage and became characterized only after booster injection. There was no activation of antibody-producing cells. An important recruitment of active macrophages was observed; this could participate to the adjuvant activity of PGL, whose at least a part of the biological activity is therefore independent of the inflammatory and/or toxic response.

Withanolides from Vassobia lorentzii.

Eight new withanolides were isolated from the aerial parts of Vassobia lorentzii and characterized by spectroscopic methods and with the aid of molecular modeling. The compounds were identified as (17S,20R,22R)-5beta,6beta:18,20-diepoxy-18-hydro xy-1-oxowitha-2,5, 24-trienolide (1); (17S,20R,22R)-18,20-epoxy-4beta, 18-dihydroxy-1-oxowitha-2,5,24-trienolide (2); (17S,18R,20R, 22R)-4beta-hydroxy-18,20-epoxy-18-methoxy-1-oxowitha-2,5, 24-trienolide (3); (17S,18S,20R,22R)-4beta-hydroxy-18, 20-epoxy-18-methoxy-1-oxowitha-2,5,24-trienolide (4); (17S,20R, 22R)-4beta-hydroxy-18,20-epoxy-1,18-dioxowitha-2,5,24-tri enolide (5); (17S,18R,20R,22R)-18,20-epoxy-18-methoxy-1,4-dioxowitha++ +-2,5, 24-trienolide (6); (17S,18S,20R,22R)-18,20-epoxy-18-methoxy-1, 4-dioxowitha-2,5,24-trienolide (7); and (17S,20R,22R)-5beta, 6beta-epoxy-4beta,18,20-trihydroxy-1-oxowitha-2,24-die nolide (8). Compounds 1 and 2 were obtained as epimeric mixtures at C-18.

Protective effect of BCG against leprosy and its subtypes: a case-control study in southern Vietnam.

A case-control study was conducted to assess the protective effect of intradermal BCG against leprosy and its subtypes in southern Vietnam. A total of 177 cases were selected with a distribution by subtypes as follows: 38 TT, 23 BT, 51 BB, 36 BL, 22 LL, and 7 indeterminate. Two controls were matched with a case for age, sex, ethnic group, socioeconomic status, and district area. The odds ratio assessing the protective effect of BCG varied from 0.44 (0.19-1.03) in the BB subtype to 3.00 (0.24-37.5) in indeterminate leprosy; whereas its overall value was 0.71 (0.45-1.10) for leprosy per se. When all borderline leprosy types were pooled, the protective effect of BCG was found significant with an odds ratio of 0.48 (0.27-0.84). In the polar forms of leprosy, TT and LL, the odds ratio was > 1 with large confidence intervals. It is possible that BCG induces a shift in the immune response to a higher level of cell-mediated immunity. When BCG vaccination is given after primary infection with Mycobacterium leprae, this shift could be the cause of an increase in the risk of the occurrence of milder and transient forms of the disease. In TT forms BCG might reinforce the preexisting subclinical immunopathological reactions, and in stable LL forms BCG might be unable to induce any protective form of immunity. These results confirm the important variability in the protection offered by BCG with respect to the different types of leprosy, and may have important implications for the design and the interpretation of vaccine trials that should take into account the respective proportions of leprosy forms observed in the study region.

Granulomatous reaction to intradermal injection of lepromin (Mitsuda reaction) is linked to the human NRAMP1 gene in Vietnamese leprosy sibships.

The Mitsuda test, which measures the specific immune response against intradermally injected lepromin, has a high prognostic value for susceptibility or resistance to the lepromatous form of leprosy. A sib-pair linkage analysis between the Mitsuda response and the NRAMP1 gene was done among 20 nuclear families with leprosy (totaling 118 sibs) from Ho Chi Minh City, Vietnam. All family subjects were genotyped for several intragenic and flanking NRAMP1 markers, leading to the definition of a fully informative NRAMP1 haplotype. Significant linkage was observed between NRAMP1 and Mitsuda reaction when considered either as a quantitative (P<.002) or as a categorical (P=.001) trait. Separate analyses among healthy and affected sibs showed evidence for linkage in both subsamples, indicating that linkage between the Mitsuda reaction and NRAMP1 is independent of leprosy status. These results support the view that NRAMP1 plays a regulatory role for the development of acquired antimycobacterial immune responses as determined by in vivo Mitsuda test reaction.

Susceptibility to leprosy is linked to the human NRAMP1 gene.

Leprosy is a debilitating infectious disease of human skin and nerves. Genetic factors of the host play an important role in the manifestation of disease susceptibility. The human NRAMP1 gene is a leprosy susceptibility candidate locus since its murine homologue Nramp1 (formerly Lsh/Ity/Bcg) controls innate resistance to Mycobacterium lepraemurium. In this study, 168 members of 20 multiplex leprosy families were genotyped for NRAMP1 alleles and 4 closely linked polymorphic markers. Highly informative haplotypes overlapping the NRAMP1 gene were constructed, and the haplotype segregation into leprosy-affected offspring was analyzed. It was observed that the segregation of NRAMP1 haplotypes into affected siblings was significantly nonrandom. This finding is consistent with the hypothesis that NRAMP1 itself is a leprosy susceptibility locus.

Complex segregation analysis of leprosy in southern Vietnam.

To investigate the nature of the genetic component controlling susceptibility to leprosy and its subtypes, 402 nuclear families were ascertained through a leprosy patient followed at the Dermatology Hospital in Ho Chi Minh City, Vietnam; 285 families were of Vietnamese origin and 117 were of Chinese origin with a higher proportion of lepromatous forms among Chinese patients. Segregation analyses were conducted using the model developed by Abel and Bonney [(1990) Genet Epidemiol 7:391-407], which accounted for variable age of onset and time-dependent covariates. Three phenotypes were considered: leprosy per se (all forms of leprosy together), nonlepromatous leprosy, and lepromatous leprosy. For each of this phenotype, analyses were performed on the whole sample and separately on the Vietnamese and the Chinese families. The results showed that a single Mendelian gene could not account for the familial distributions of leprosy per se and its two subtypes in the whole sample. However, these results were different according to the ethnic origin of the families. In the Vietnamese subsample, there was evidence for a codominant major gene with residual familial dependences for the leprosy per se phenotype, and borderline rejection of the Mendelian transmission hypothesis for the nonlepromatous phenotype. In Chinese families, strong rejection of Mendelian transmission was obtained in the analysis of leprosy per se, and no evidence for a familial component in the distribution of the nonlepromatous phenotype was observed. For the lepromatous phenotype, the discrimination between models was poor, and no definitive conclusion could be reached. Referring to immunological data, we suggest that these results could be explained by a heterogeneity in the definition of the lepromatous phenotype. It is likely that progress in the understanding of the genetic components involved in the expression of leprosy will come from a better definition of the phenotype under study, and immunological studies are ongoing in this population to investigate this hypothesis.