[Acquired C1 esterase inhibitor deficiency via bradykinin-mediated angioedema: Four cases]. / Angiœdème bradykinique par déficit en C1 Inhibiteur acquis : quatre cas.

BACKGROUND: Acquired C1-esterase inhibitor (C1-INH) deficiency angioedema (C1-INH-AAE) is a form of bradykinin-mediated angioedema. This rare disorder is due to acquired consumption of C1-INH, hyperactivation of the classic pathway of human complement, and potentially fatal recurrent angioedema symptoms. Clinical symptoms of C1-INH-AAE are very similar to those of hereditary angioedema (HAE) but usually appear after the fourth decade of life and induce abdominal pain less frequently. Laboratory tests are essential in establishing the diagnosis with low levels or abnormal structure and function of C1-INH. Most patients present C1-INH autoantibodies. Furthermore, C1q is reduced in AAE, contrary to HAE. The long-term prognosis is determined by associated hematologic malignancies. PATIENTS AND METHODS: We report 4 cases of C1-INH-AAE associated with lymphoproliferative disorders referred to the Reference Centre for Angioedema of Besançon, France. The patients were aged between 60 and 77 years. C1 INH antibodies were found in three patients. Symptoms were triggered by angiotensin-converting enzyme (ACE) inhibitors or angiotensin II receptor blockers (ARBs) in 3 patients. Hematologic malignancy was present at diagnosis (one case of chronic lymphoid leukemia) or was diagnosed during follow-up (one case of indolent marginal zone non-Hodgkin lymphoma and two cases of monoclonal gammopathy). DISCUSSION: C1-INH-AAE induced by ACE inhibitors or ARBs may be associated with hematologic malignancies. This form of revelation does not necessarily indicate a diagnosis of ACE or ARBs angioedema, and screening should therefore be performed for C1 Inh and C1q. An underlying hematologic malignancy should be routinely sought and the long-term prognosis determined.

[Immune monitoring of kidney transplant recipients: can markers predictive of over-immunosuppression be identified?]. / Surveillance biologique des patients transplantés rénaux : vers une prévision des complications associées à l'immunosuppression ?

While the use of nonspecific immunosuppressive drugs has significantly reduced the incidence of acute graft rejection, the benefits of such therapies on chronic rejection and overall long-term graft survival are uncertain. Persistent excessive immunosuppression after immunosuppressive drug treatment is associated with long-term toxicity including increased incidence of cancers, severe infectious complications and metabolic diseases (for example, diabetes, atherosclerosis). One of our team's aims is to identify immunological factors that can predict such toxicities. We have previously demonstrated that CD4T cell cytopenia was correlated with high risk of cancers and infections as well as atherosclerosis in renal transplant recipients. Now, we are investigating the mechanisms involved in CD4T cell cytopenia. We are also exploring how inflammation and cells from the innate immunity influence the complications associated with kidney transplantation. This was performed through the analysis of gene polymorphism on TLR-4, NOD2/CARD15 receptors and IL-6 promoter and correlation with transplantation outcome. We already correlated IL-6 promoter gene polymorphism at position -174 with new-onset diabetes after transplantation in overweight patients. Identification of gene polymorphisms or factors associated with complications after transplantation may help physicians to determine high-risk recipient profiles and optimize pre- and post-transplantation treatment strategies.

Intravenous apoptotic spleen cell infusion induces a TGF-beta-dependent regulatory T-cell expansion.

Apoptotic leukocytes are endowed with immunomodulatory properties that can be used to enhance hematopoietic engraftment and prevent graft-versus-host disease (GvHD). This apoptotic cell-induced tolerogenic effect is mediated by host macrophages and not recipient dendritic cells or donor phagocytes present in the bone marrow graft as evidenced by selective cell depletion and trafficking experiments. Furthermore, apoptotic cell infusion is associated with TGF-beta-dependent donor CD4+CD25+ T-cell expansion. Such cells have a regulatory phenotype (CD62L(high) and intracellular CTLA-4+), express high levels of forkhead-box transcription factor p3 (Foxp3) mRNA and exert ex vivo suppressive activity through a cell-to-cell contact mechanism. In vivo CD25 depletion after apoptotic cell infusion prevents the apoptotic cell-induced beneficial effects on engraftment and GvHD occurrence. This highlights the role of regulatory T cells in the tolerogenic effect of apoptotic cell infusion. This novel association between apoptosis and regulatory T-cell expansion may also contribute to preventing deleterious autoimmune responses during normal turnover.

Polymeric Ig receptor expression in hepatocellular carcinoma.

The cellular localisation of the polymeric Ig receptor (pIg-R) and carcinoembryonic antigen (CEA), hepatic and biliary cell markers, were investigated in patients with hepatocellular carcinoma (HCC) and high serum levels of secretory component. Serum SC were increased 6-20-fold in 8 HCC patients compared with normal subjects. Serum free SC was positively correlated bilirubin (r = 0.95, P less than 0.04). In normal liver tissue, cytokeratin (CK) 8 and 18 were localised in hepatocytes and biliary cells while pIg-R and CK 19 expression was restricted to biliary cells. In tumoral liver tissue, malignant cells expressed CK 8 and 18 weakly; pIg-R and CK 19 were not detected in tumoral cells. CEA was expressed by biliary cells in normal and proliferating ducts. In peritumoral fibrosis, proliferating biliary cells were strongly stained by anti-cytokeratins and anti-pIg-R antibodies. In one case, pIg-R was localised in isolated cells close to fibrosis without co-staining of anti-CK 19. Thus increased serum SC is not associated with pIg-R expression by tumoral cells, and pIg-R may be considered an additional marker of biliary cells. High SC might be explained either by reflux from bile to serum and/or release of unbound SC from the vascular pole of non-functional, proliferating biliary structures.

High-risk papillomavirus infection is associated with altered antibody responses in genital tract: non-specific responses in HPV infection.

In order to gain more information about local humoral immune responses to HPV infection, we quantified IgG, IgM, secretory-IgA (S-IgA), and total-IgA by ELISA, and lysozyme and lactoferrin by TR-IFMA, in cervical and cervicovaginal secretions of 40 healthy women and 28 high-risk HPV infected patients (11 were HPV16+). IgG, total-IgA, and S-IgA concentrations in cervicovaginal secretions (p < 0.0001) and high IgG and total-IgA concentrations (p < 0.001 and p < 0.01, respectively) in endocervical secretions were significantly higher in HPV+ patients than in the healthy group. Since the S-IgA/total-IgA ratio was significantly lower in cervicovaginal (7.5%) and endocervical secretions (36.5%) in HPV+ women compared to the control group (p < 0.003 and p < 0.001, respectively), HPV could be responsible for an increase in local production of non-secretory IgA (monomeric and dimeric forms). IgG and total-IgA concentrations in cervicovaginal and endocervical secretions fell in the same general percentage range in both HPV16+ and HPV+ groups (80% and 15%, respectively). However, the S-IgA/total-IgA ratio was much lower in HPV16+ than in HPV+ women, in both cervicovaginal secretions (3.4%) (p < 0.003) and in endocervical secretions (23.3%) (p < 0.001). Innate immunity proteins and local S-IgA response could not stop the spread of HPV infection in spite of high lysozyme and lactoferrin concentrations. HPV16+ disturbed the local humoral immune system, which could partly explain its low clearance.

Genital human papillomavirus infection among women recruited for routine cervical cancer screening or for colposcopy determined by Hybrid Capture II and polymerase chain reaction.

The purpose of this study was to evaluate the clinical use of the Hybrid Capture (HC)-II system for the detection of human papillomavirus (HPV) DNA to identify women at risk of progression to high grade squamous intraepithelial lesions (HGSIL) and carcinomas by differentiating low risk (LR) HPV types (6, 11, 42, 43, 44) and high/intermediate risk (HR) HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68). Five hundred and ninety-six women were enrolled in the study. Among them, 466 attended the hospital for routine cytologic screening and 130 were referred for colposcopy because of an abnormal Pap smear. The presence of HPV DNA was tested in cervical samples collected with the Digene Cervical Sampler in Digene Specimen Transport Medium (Digene Corporation, Silver Spring, MD, U.S.A.) using the HC-II assay. Results were compared with those obtained by polymerase chain reaction (PCR) using the MY09-MY11 primers followed by several hybridizations with specific probes. The overall HPV positivity was 32.9% by HC-II and 37.8% by PCR. Among cytologically normal smears, 19.5% were positive by HC-II (14.3% HR) and 25.1% by PCR. Of the atypical squamous cells of undetermined significance samples, 52.9% were positive by HC-II (41.1% HR) and 55.9% by PCR. Of the low grade SIL, 64.5% were positive by HC-II (59.4% HR) and 68.7% by PCR. The HPV positivity rate was found identical by both techniques in high grade smears (81.6%) and squamous cervical carcinomas (100%). By using PCR as the reference method, the sensitivity of HC-II was higher among women with abnormal cytology than with normal cytology (87.3% vs. 70%). Specificity was 80.8% and 97.5%, respectively. In summary, these results indicate that the HC-II method and MY-PCR identified nearly equivalent prevalences of HPV in cervical smear specimens.

Light chains of immunoglobulins in human secretions.

Immunoglobulin kappa light chains are predominant in normal human serum and a kappa/lambda ratio of 1.7 to 2 has been reported. However, little is known of the partition of light chains in secretions. The levels of IgA, kappa and lambda were assayed by nephelometry in a series of secretions and in normal human serum. Although kappa chains remained predominant, the different secretions studied could be classified according to their kappa/lambda ratio. In saliva (P < 0.001), nasal fluid (P = 0.01) and tears (P = 0.04) the kappa/lambda ratio was significantly lower than in serum. Conversely, higher kappa/lambda ratios were obtained in gastric juice (P = 0.005) and hepatic bile (P = 0.004) and no significant difference was noted between serum and gall bladder bile (P = 0.62). The lower ratios were all observed in fluids produced by glands surrounded by lymphoid tissue included in the mucosae-associated lymphoid tissue. These data strengthen previous observations that suggest a preferential production of lambda chains in human mucosae.

Triazinic herbicide determination by gas chromatography-mass spectrometry in breast milk.

A solid-phase extraction procedure using a graphitized carbon black cartridge for extraction and cleaning of a series of five triazines (atrazine, deethylatrazine, deisopropylatrazine, ametryne and prometryne) from breast milk samples was developed. Using a chemometric methodology, the optimisation of both the analysis time and the triazinic herbicide separation by gas chromatography-mass spectrometry (GC-MS) was then carried out with only 18 experiments. Detection and quantification limits for 1ml breast milk sample were, respectively, 0.3 and 1 ppb for each studied compound. The variation coefficients were less than 5% over the concentration range from 1 to 100 ppb. The accuracy was between 98.63 and 104.62% for each triazinic herbicide. The recovery was between 58.64 and 63.22% for the concentration range from 1 to 100 ppb for each triazinic herbicide. The assay was successfully applied to the analysis of several breast milk samples.

Serum secretory immunoglobulins in ankylosing spondylitis.

Humoral mucosal immunity may be implicated in pathophysiology of ankylosing spondylitis (AS). The aim of the study was to evaluate serum levels of IgA, IgM and secretory IgA (sIgA), secretory IgM (sIgM) as well as free secretory component (FSC) in patients with AS compared to controls and rheumatoid arthritis (RA) patients. Levels of sIgA, sIgM and FSC were measured with a specific ELISA in 37 AS patients, 45 controls and 27 RA. The results were as follows: Serum levels of IgA were higher in AS vs controls and in RA vs controls (p = 0.01). Levels of sIgA were higher in AS vs controls (p = 0.01), but higher in RA vs AS (p = 10(-4)). There was no difference of sIgM in AS vs controls, FSC levels were higher in AS vs controls, and higher in AS patients with elevated CRP. In view of elevated FSC, this increase of sIgA in AS may have been due to excessive production of mucosal IgA after bacterial stimulation according to the current hypothesis of the disease.

Human papillomavirus detection by non isotopic in situ hybridization, in situ hybridization with signal amplification and in situ polymerase chain reaction.

Classical in situ hybridization (ISH) with biotinylated probes makes it possible to detect and localize human papillomavirus (HPV) nucleic acid sequences in cytological and histological materials. This method is however of limited value in the detection of a few copies of the virus. Moreover the specificity of such a technique is not always convincing when ISH signals are small and/or of low intensity. Recently, much attention has been focused on the utility of the in vitro polymerase chain reaction (PCR) and especially on PCR-single strand conformation polymorphism (SSCP) to amplify small amounts of viral DNA with accurate hybrid specificity. But the latter method requires nucleic acid extraction and tissue destruction. Thus, correlation between the PCR results and histological findings is not possible. Hence, the aim of our current study was to apply to HeLa cells and cervical formalin-fixed and paraffin-embedded biopsies, a novel procedure of ISH signal amplification, the catalyzed signal amplification (CSA). Such a procedure is based on the deposition of streptavidin-horseradish peroxidase catalyzing the deposition of biotinylated tyramide molecules on the location of the probed target. The biotin accumulation is then detected with streptavidin peroxidase and diaminobenzidine. The results were compared with those obtained by direct and indirect in situ PCR. The catalysed signal amplification successfully increased the sensitivity and efficiency of ISH for the detection of rare sequences in HPV infected cells and histological materials. Such a method was found simpler and faster than in situ PCR and tissue morphology was better preserved.

[IgA and its different molecular forms in the mesenteric, portal and peripheral venous blood in man]. / L'IgA et ses différentes formes moléculaires dans le sang veineux mésentérique, portal et périphérique chez l'homme.

The aim of this study was to assess the role of mesenteric blood in polymeric IgA (p-IgA) and IgA2-transport from the intestinal mucosa into plasma and the role of the liver in the clearance of these molecular forms of IgA. The concentrations of IgA, p-IgA and IgA2 were measured in mesenteric, splenic, portal, and hepatic veins of 7 control subjects without liver disease and in portal and peripheral veins of 4 patients with alcoholic cirrhosis. In control subjects, the concentration of the different molecular forms of IgA were not significantly different in mesenteric and in splenic vein. No significant decrease of IgA concentrations was observed in hepatic vein, as compared with portal vein. In cirrhotic patients IgA concentrations were significantly higher than in control subjects, but concentrations of IgA, p-IgA and IgA2 were not different in portal and peripheral blood. These results show that mesenteric vein is not a major way for p-IgA and IgA2 from the gut lamina propria to plasma, and suggest that the origin of a significant part of these molecular forms of IgA could be peripheral lymph-nodes more than gut-associated-lymphoid-tissue. The absence of significant clearance of p-IgA by the liver in normal subjects suggests that abnormalities of hepato-biliary transport of p-IgA is not responsible for the increased IgA levels observed in cirrhotic patients.

[The biology of papillomavirus infections. III. Immune response]. / Biologie des infections à papillomavirus. III. Réponse immunitaire.

Infection with the human papillomaviruses, especially with oncogenic HPVs increases the risk for development of precancerous and cancerous lesions of the cervix. The immune response of the host is likely to be an important factor in determining regression or progression of papillomaviruses-associated lesions. Systemic IgG and IgA response is classically associated with current or past papillomavirus infections. A deficiency in local cellular immune response is however frequently observed and linked to a decrease of cytokine synthesis by infected cells, a reduction or loss of MCH I molecules and a defect in antigen presentation to cytotoxic T lymphocytes. Although secretory immunoglobulins are generated locally in response to papillomavirus infections, humoral immunity in the female genital reproductive tract seems to be inefficient. The papillomavirus infections would lead to a decrease in cellular immunity which could be favourable to viral latency and/or precancerous and cancerous lesion development.

[Biology of the papillomavirus infections: V. Vaccine developments]. / Biologie des infections à papillomavirus: V. Perspectives vaccinales.

Several genotypes of human papillomaviruses (HPV) are recognised as aetiologic factors for cervical cancer, and viral DNA account for more 99% of cases. Thus, prevention of HPV infection by, for example, types 16 and 18, should reduce the world-wide incidence of cervical cancer. Many strategies are being developed for the control of HPV-associated lesions of the uterine cervix: prophylactic vaccines which elicit neutralizing antibodies to prevent HPV infection, and therapeutic vaccines which induce a T-cytotoxic response to early viral oncoproteins. Experimental trials are being conducted to test mucosal immunization with an ideal antigen delivery system. Vaccination strategies elicit a protective antibody response in animal species, but in humans, strategies which are likely to be effective in the control of HPV-associated preneoplastic and neoplastic lesions of the uterine cervix are still under investigation.

[Biology of papillomavirus infections. IV. Sero-epidemiological data]. / [Biologie des infections à papillomavirus: IV. données séro-épidémiologiques.

Serological assays using synthetic peptides or recombinant proteins to identify specific HPV infection either fail to correlate with the type of infection or are only able to identify a small percentage of infected individuals. But genetically engineered Virus Like Particles (VLP) seem useful to develop Elisas for serological diagnosis of HPV infection and antibodies to VLP appear to correlate well with HPV DNA presence. The levels of secretory immunoglobulins in genital secretions would provide a better indicator of HPV infection, but reproducibility and standardization of the detection methods are unresolved. The clinical relevance of serologic responses is still questionable, since frequency and titer of several types of antibodies generated against HPV show a great variability which is dependent on the HPV type specificity, on the recognized epitopes and on the type of samples. However the detection of neutralizing antibodies associated with disease recurrence and antibodies raised against HPV16 E6 and E7 peptides found to react with sera of cancer patients, needs to pay attention.

[Immunity of the female genital tract mucosa and mechanisms of papillomavirus evasion]. / Immunité muqueuse du tractus génital féminin et mécanismes d'évasion des papillomavirus.

Human papillomaviruses (HPV) are responsible for many cutaneous and mucosal lesions. Some viral genotypes are considered to be the causal agents of cervical cancer. Natural genital HPV infection seems to be poorly immunogenic because of its nonproductive and noninflammatory characteristics and also because of the different mechanisms developed by the virus to counteract the immune response. Knowledge of the immune system organization and its regulation in the human female genital tract needs to be clarified. It is mostly "programmed" to ensure a humoral response. Nevertheless, secretory IgA, that are particularly efficient for anti-infectious mucosal immunity are poorly present in physiological vaginal secretions. These distinctive features could explain part of the relative immune deficiency against HPV. Moreover, reduction or loss of MCH1 molecules and a defect in antigen presentation to cytotoxic lymphocytes could in part explain the cytotoxicity deficiency. There is however clear evidence that cellular immune response plays a major role in the control and course of HPV infection. This response varies according to the grade of the lesion and to the oncogenic potential of the infecting HPV. A deficiency in induction of cellular cytotoxicity mechanisms seems to be involved in the persistence of HPV infection and so in carcinogenesis. Finally, worldwide cervical cancer incidence (5000000 new cases per year) warrants effective vaccine developments. Two strategies, one preventive and one therapeutic, are now under study. Vaccine adjustments are based first on humoral immunity induction with production of neutralizing antibodies for prophylaxis and second on cellular immunity induction to kill cells with viral oncoprotein expression for therapy.

[Serum and salivary immunoglobins A in atopic dermatitis. Prospective and comparative case control study]. / Immunoglobulines A sériques et salivaires dans la dermatite atopique. Etude prospective et comparative témoins-malades.

UNLABELLED: IgA system has been poorly studied in patients with atopic dermatitis (AD). Previous studies have showed that a transient serum IgA deficiency in infancy could lead to atopic disease. In addition, decrease in salivary IgA has been demonstrated in patients with AD. The purpose of our work was to study the IgA system both in serum saliva in patient with AD. PATIENTS AND METHOD: We conducted a controlled prospective study from January 1994 to May 1996. 46 patients with AD and 52 healthy volunteers matched for sex and age were included. Atopic patients fulfilled at least three major and three minor features defined by Hanifin and Rajka. None above atopic criteria were present in the control group. Saliva was collected using a small cylinder of a cotton-wool-like substance (Salivette) kept in the buccal fold. Serum and saliva samples were assayed for IgA using standard nephelometric method and time-resolved immunofluorometric assay. Secretory IgA were assayed by a sandwich-type enzyme linked immunosorbent assay. Blood eosinophils and serum IgE were also evaluated. RESULTS: IgA and secretory IgA were detected in all serum and saliva collected. No statistically significant difference were observed in serum or in saliva for both IgA and secretory IgA between patients with AD and controls. As expected, blood eosinophils and serum IgE were significantly increased in patients with AD. DISCUSSION: None patients (atopic or control) exhibited IgA deficiency. Although no statistically significant, a trend to higher concentrations of serum and salivary IgA was observed in patients with AD suggesting a stimulation of mucosa-associated lymphoid tissue in these patients.

[Imidazo(2,1-b)thiazole XII derivatives. Synthesis and research of immunoactivity in vitro on human T lymphocyte of 3-aroylmethyl and 2-aroyl-3-methyl(aryl)-5,6-dihydroimidaz o(2,1-b)thiazoles]. / Dérivés de l'imidazo[2,1-b]thiazole XII. Synthèse et recherche d'une immunoactivité in vitro sur le lymphocyte T humain de 3-aroylméthyl et de 2-aroyl-3-méthyl(aryl)-5,6-dihydroimidazo[2,1-b]thiazoles.

To estimate the influence of aryl group position on the immunostimulant properties of imidazo[2,1-b]thiazole derivatives, several compounds were obtained and tested, versus tetramisole hydrochloride, on the mobilisation of CD2 receptor by human T lymphocyte. The synthesis use the action of monobrominated beta-diketones on the 2-mercaptoimidazoline. So, 1-aryl-4-bromobutane-1,3-diones lead to 3-aroylmethyl-5,6-dihydroimidazo[2,1-b]thiazoles 4. Same, 1-aryl-2-bromobutane-1,3-diones and 1,3-diaryl-2-bromopentane-1,3-diones give respectively 3-aroyl-2-methyl-5,6-dihydroimidazo[2,1-b]thiazoles 5 and 3-aroyl-2-aryl-5,6-dihydroimidazo[2,1-b]thiazoles 6. Better yields are obtained when the reaction presents two steps. The first one, realized in acetone at room temperature, leads to an intermediate S-substituted 4,5-dihydroimidazole which, in second step, is cyclized in imidazo[2,1-b]thiazole compound via an unisolated carbinolamine. This one explains the univocal formation of 5 derivatives and the feasible blending in case of 6. The immunoactivity of several imidazothiazoles 4, 5 and 6 is lower that them of 6-aryl substituted compounds and particularly that the levamisole which is the reference product in this series.

[Immunomodulator structure-activity relationships: contribution of molecular modeling]. / Relations structures-activités des immunomodulateurs. Apport de la modélisation moléculaire.

Molecular modeling used to compare 64 immunostimulant compounds with pyrrolie quinolein or purine nuclei has pointed out that a common spatial structure is found in most of the active compounds. An additional study of immunostimulants (levamisole, muramyldipeptide) or immunosuppressive molecules (rapamycin) was performed. A common pharmacophore was found on every studied compound. It was composed of three neighboring electroattractive atoms and a further fourth atom. The favorable conformation of rapamycin for immunosuppressive action, which is not the more stable conformation, could explain the loss of its activity, or those of related macrolides, when some minor chemical modifications are tested. These findings validate the proposed concept and provide a view of the mechanism of action of most of the immunomodulator compounds for preparing novel compounds

Phosphatidylserine-expressing cell by-products in transfusion: A pro-inflammatory or an anti-inflammatory effect?

Labile blood products contain phosphatidylserine-expressing cell dusts, including apoptotic cells and microparticles. These cell by-products are produced during blood product process or storage and derived from the cells of interest that exert a therapeutic effect (red blood cells or platelets). Alternatively, phosphatidylserine-expressing cell dusts may also derived from contaminating cells, such as leukocytes, or may be already present in plasma, such as platelet-derived microparticles. These cell by-products present in labile blood products can be responsible for transfusion-induced immunomodulation leading to either transfusion-related acute lung injury (TRALI) or increased occurrence of post-transfusion infections or cancer relapse. In this review, we report data from the literature and our laboratory dealing with interactions between antigen-presenting cells and phosphatidylserine-expressing cell dusts, including apoptotic leukocytes and blood cell-derived microparticles. Then, we discuss how these phosphatidylserine-expressing cell by-products may influence transfusion.