High prevalence of latent tuberculosis infection in autoimmune disorders such as psoriasis and in chronic respiratory diseases, including lung cancer.

The early diagnosis and treatment of individuals harboring M. tuberculosis is key to ensuring the effectiveness of health programs aimed at the elimination of tuberculosis (TB). Monitoring for TB also has other important health care implications for the related immune pathology caused by the chronic inflammatory response to M. tuberculosis. Moreover, the recent introduction of biologic therapies for the treatment of several immune-mediated inflammatory diseases has shown unexpected high frequencies of reactivation of latent TB. The present cross-sectional study is aimed at estimating the prevalence of latent tuberculosis infection (LTBI) in different groups of subjects, either undergoing a routine program of screening for TB or a clinical monitoring of autoimmune or lung disorders, by analyzing their immune response in vitro to a pool of different M. tuberculosis antigens through an IFN-gamma-release assay (IGRA). We consecutively tested 1,644 subjects including health care workers (931), healthy immigrants from different countries (93), patients with a diagnosis of psoriasis (405), patients with lung inflammatory disease (60) or lung neoplasia (32) and a group of HIV-1 infected Italian subjects (120). The prevalence of IGRAs positive responses among health care workers was 8.9 percent. In comparison, significantly higher frequencies were found in healthy immigrant subjects (33.3%), similar to those found in inflammatory broncho-pneumopathies (34.5%) or lung cancer (29.6%). Interestingly, an unexpected high prevalence was also found in patients affected by psoriasis (18.0%), while HIV-infected subjects had values comparable to those of health care workers (10.8%). An age cut-off was determined and applied for each group by receiver operating characteristic (ROC) curves in order to perform the statistical analysis among age-comparable groups. Multivariate analysis showed that the age and clinical conditions such as having a diagnosis of psoriasis or a lung inflammatory disease were independent risk factors for developing an IGRA positive response. This study highlights an unprecedented high prevalence of IGRA positive responses among patients affected by psoriasis and emphasizes the need for a preliminary assessment of LTBI before the administration of any biologic therapy based on cytokine antagonists such as anti-TNF-alpha. Moreover, screening for LTBI should be routinely performed in the presence of a chronic pulmonary disease.

Epithelial lining fluid neutrophil-gelatinase-associated lipocalin levels in premature newborns with bronchopulmonary dysplasia and patency of ductus arteriosus.

Patency of the ductus arteriosus (PDA) and bronchopulmonary dysplasia (BPD) development represent severe affections for premature newborns, therefore the research of early markers for these two conditions is really important. The aim of this study is to analyze epithelial lining fluid (ELF) Neutrophil-gelatinase-associated lipocalin (NGAL) levels for prediction of lung injury or possible involvement of this molecule in PDA. Only scarce and contrasting results have previously been published in this field. In contrast, this molecule, included in a large macromolecular complex together with matrix metalloproteinase-9 (MMP-9), is considered an acceptable marker of infectious/inflammatory processes, cancer monitoring and induction of apoptotic pathway. NGAL was detected in 28 pre-term newborns by means of a commercially available kit in bronchoalveolar lavage fluid (BALF). The results have been corrected to ELF levels, by the urea method, to eliminate bias due to BALF collection. ELF NGAL levels were found significantly increased both in infants developing BPD or in those affected by PDA. By means of multivariate logistic regression analysis the significances were confirmed after adjusting for possible interfering variables such as gestational age and concomitant presence of both PDA and BPD. Our results stress the involvement of NGAL in the mechanisms leading to BPD and also suggest a possible association with PDA, which is often linked to prematurity and BPD development, probably due to the involvement of inflammatory and angiogenetic processes in both pathologies.

Slight association between type 1 diabetes and "ff" VDR FokI genotype in patients from the Italian Lazio Region. Lack of association with diabetes complications.

BACKGROUND: Vitamin D receptor (VDR) mediates the effects of vitamin D. Our paper evaluates the FokI and BsmI VDR genotypes in 246 Caucasian (Italian from Lazio Region) T1DM patients compared with 246 Caucasian healthy controls, sharing age and gender and regional provenience with the patients. In addition, T1DM patients without complications were compared with those carrying three complications. METHODS: Genotyping has been obtained by RFLP-PCR technique. RESULTS: A slight significant association of T1DM with FokI homozygous "f" genotype was observed. No association was observed with the presence of multiple complications by a multivariate analysis. CONCLUSION: T1DM patients showed slightly increased prevalence of "ff" VDR genotype.

Increased levels of IGF-1 and beta2-microglobulin in epithelial lining fluid of preterm newborns developing chronic lung disease. effects of rhG-CSF.

UNLABELLED: Insulin-like growth factor-1 (IGF-1) is involved in regulating the Th-1/Th-2 balance, favoring the development of the Th-2 compartment which enhances fibrosis, one of the main characteristics of Chronic Lung Disease (CLD) in premature newborns. Limited data is available concerning a possible association between early epithelial lining fluid (ELF) concentrations of IGF-1 (total and free forms), IGF-binding protein-3 (IGFBP-3), beta2-microglobulin and subsequent development of CLD in preterm neonates. If neutropenic, preterm neonates are frequently treated with recombinant human granulocyte colony stimulating factor (rhG-CSF). The objective of the study was to correlate ELF concentrations of IGF-1 and beta2 microglobulin during the first week of life both in non-neutropenic and in rhGCSF-treated neutropenic preterm neonates, with subsequent development in CLD. Thirty preterm neonates with Respiratory Distress Syndrome (6 with neutropenia) were studied. Eleven out of 24 non-neutropenic preterm infants (46%) and all of the six neutropenic subjects (100%) developed CLD. With the exception of first day values, there was a clear similarity in the behaviors of assayed molecules between non-neutropenic and neutropenic patients developing CLD. Non-neutropenic patients without CLD showed significantly lower values of free IGF-1 and beta2M both on days 1 and 3. Total IGF-I and cell counts were different only on the 3rd day. CONCLUSIONS: 1) the mechanisms leading to CLD might be mediated by high levels of IGF-family molecules soon after birth 2) beta2M could be a marker of increased bronchoalveolar lavage fluid cellularity with potential inflammatory properties 3) G-CSF treatment induces an increased synthesis of IGF-1 molecules by cells recruited in the lung, with possible enhancement of the fibrogenic mechanisms.

c-MYC deregulation is involved in melphalan resistance of multiple myeloma: role of PDGF-BB.

Oncogenes are important regulators of cancer growth and progression and their action may be modulated by proteins of the growth factor family, such as angiogenic cytokines, known to be strongly involved in neoplastic evolution. Reciprocal interactions between oncogenes and angiogenic modulators may represent, in haematological neoplasms, including multiple myeloma (MM), a possible mechanism of drug resistance. The aim of this work is to investigate in vitro and in vivo whether or not c-myc deregulation is involved in the melphalan resistance elicited by myeloma patients and consequently to clarify the role of the angiogenic factor PDGF-BB in modulating c-myc protein expression. Fifty-one MM patients on chemotherapy with melphalan were analyzed for structural alterations of the c-myc gene, c-Myc protein expression, as well as for serum PDGF-BB release. For the in vitro study, two M14-derived established cell clones, differing for the c-Myc protein expression (c-Myc low -expressing or constitutively expressing clones) were used. Our results show that PDGF-BB is able to up-regulate Myc expression and reduce melphalan sensitivity of tumor cell clones, constitutively expressing c-myc gene product. In addition, down-regulation of c-Myc protein induces the expression of PDGF-beta receptor molecules and reduces PDGF-BB release. In agreement with these results, in vivo data show that melphalan-resistant MM patients present overexpressed c-Myc protein and higher serum PDGF-BB receptor levels compared to minor responding patients.

Prevalence of human papilloma virus type 5 DNA in lesional and non-lesional skin scales of Italian plaque-type psoriatic patients: association with disease severity.

Human papilloma virus type 5 (HPV-5) has been associated closely with psoriatic skin in Polish patients, while findings from other countries have indicated a more limited prevalence. The results of the present study, in which a type-specific nested PCR was used, indicated that scales of plaque-type psoriatic skin from 54 Italian patients had a high prevalence (74.1%) of HPV-5 DNA in lesional areas, and a reduced prevalence (33.3%) in non-lesional skin (33.3%), compared to 0% of 20 healthy subjects and 3.6% in the lesional areas of 28 patients with various other dermatological diseases. Individuals negative for HPV-5 DNA had a less severe disease. No correlation was found between the presence of HPV DNA and a patient's age or sex. The data demonstrated a statistically significant association between psoriasis and HPV-5, although results in other geographical areas suggest variable virus spread or ethnic variation in virus colonisation.

Post-treatment changes of six cytokines in active pulmonary tuberculosis: differences between patients with stable or increased fibrosis.

SETTING: Little information is available regarding the relationship between the fibrotic evolution of pulmonary tuberculosis (PTB) and cytokine levels in human bronchoalveolar lavage fluid (BALF) and serum. OBJECTIVE: To evaluate correlations between profibrotic cytokine levels and post-treatment lung fibrotic evolution. DESIGN: BALF and serum amounts of pro- or anti-inflammatory cytokines were obtained by ELISA before and 6 months after the start of anti-tuberculosis chemotherapy in 13 subjects with PTB. BALF levels were recalculated as ELF (epithelial lining fluid) levels by the urea method. High resolution computed tomography (HRCT) of both lungs was performed at the same time. RESULTS: When comparing pre- and post-treatment radiological data, the scores for 2-10 mm nodules, consolidation and fibrosis presented significant differences (P < 0.05). Concomitantly, pre-treatment vs. 6 month concentrations of ELF IFN-gamma and TNF-alpha were decreased (P < 0.05), while those of IL-4 and IL-10 were increased (P < 0.012). At serum level, IFN-gamma decreased, as did TNF-alpha, TGF-beta1 and PDGF-BB (P < 0.05). When the patients were subdivided into two groups, 1) stable or 2) increasing HRCT fibrosis score, significant increases in the second group were observed for ELF/ serum values of TGF-beta1 and ELF PDGF-BB (P < 0.05) at 6 months post-treatment. Only serum TGF-beta1 values were significantly associated with the same group before treatment.

T-cell apoptosis induced by granulocyte colony-stimulating factor is associated with retinoblastoma protein phosphorylation and reduced expression of cyclin-dependent kinase inhibitors.

Peripheral blood progenitor cells (PBPC) mobilized by granulocyte colony-stimulating factor (G-CSF) promptly engraft allogeneic recipients after myeloablative chemotherapy for hematologic malignancies. Surprisingly, no exacerbation of acute graft-vs-host disease has been observed despite a 10-fold higher T-cell content in PBPC compared with bone marrow allografts. Because G-CSF can suppress T-cell proliferation in response to mitogens and enhance their activation-induced apoptosis, we examined the molecular mechanisms underlying G-CSF-induced immune dysfunction. Normal allogeneic lymphocytes were challenged with phytohemagglutinin in the presence of serum collected after G-CSF administration (postG) to healthy PBPC donors, and the expression of key components of the cell cycle and apoptotic machineries was investigated by flow cytometry and Western blotting. Lymphocyte stimulation was associated with collapse of mitochondrial transmembrane potential, hypergeneration of reactive oxygen intermediates, and activation of caspase-3 and DNA fragmentation. Lymphocytes were arrested in a G(1)-like phase of the cell cycle, as measured by G(1)-phase cyclin expression and bromodeoxyuridine (BrdUrd) incorporation. Cell tracking experiments confirmed the occurrence of a lower number of population doublings in postG compared with preG cultures. Unexpectedly, the phosphorylation state of the protein encoded by the retinoblastoma susceptibility gene (pRB) was unaltered in postG cultures, and the inhibition of cell cycle progression occurred without the recruitment of the cyclin-dependent kinase inhibitors p15(INK4B), p16(INK4A), and p27(Kip1). We eventually evaluated the ability of antioxidant/cytoprotectant agents to prevent the G-CSF-induced mitochondrial dysfunction and inhibition of cell cycle progression. Of interest, both N-acetylcysteine and amifostine reduced apoptotic cell death by 45% on average, inhibited the activation/processing of caspase-3, and increased BrdUrd incorporation in postG cultures. Based on these experimental findings, a model is proposed in which T-cell activation in the presence of serum immunoregulatory factor(s) induced by G-CSF is associated with a molecular phenotype mimicking the G(1)-S transition and consisting of pRB phosphorylation, lack of CDKI recruitment, and reduced cyclin-E expression. The putative relationship between lymphocyte mitogenic unresponsiveness and apoptosis induction would occur at the level of key molecules shared by the cell cycle and apoptotic machineries. Whether the G-CSF-mediated modulation of lymphocyte functions in vitro is beneficial in transplantation medicine remains to be determined.

Unidirectional budding of HIV-1 at the site of cell-to-cell contact is associated with co-polarization of intercellular adhesion molecules and HIV-1 viral matrix protein.

OBJECTIVES: To explore the possibility that HIV-1 budding and cellular adhesion molecules co-polarize at cell-to-cell contact sites. To investigate the incorporation of host-cell-derived adhesion molecules into HIV-1. METHODS: The cellular sites involved in HIV-1 budding were examined by transmission electron microscopy. Single and double immunocytochemistry staining was used to evaluate the cellular distribution of the viral matrix protein and adhesion molecules. Quantitative flow cytometry was used to measure the cellular expression of adhesion molecules. An immunocapture technique was used to measure the presence of cell-derived proteins on HIV-1. The captured virus was measured by a p24 antigen assay. The infectivity of virus captured by monoclonal antibodies was tested by measuring the virus antigen yield in supernatants after the addition of sensitive cells. RESULTS: Released and budding HIV-1 was mainly localized at the cell-to-cell contact regions. This feature was consistent with a polarized staining for the virus matrix protein p18 at cell-to-cell contact regions. Intercellular adhesion molecules (ICAM)-1 in HIV-1-infected cells were polarized on both isolated cells and syncytia, co-localizing with HIV-1 matrix protein. HIV-1 incorporated all the adhesion molecules expressed by the host cells, although without quantitative correlation with their cellular expression. CONCLUSIONS: HIV-1 is released at cell-to-cell membrane contact sites. Both ICAM-1 and virus matrix protein co-polarized on isolated cells and syncytia at the sites involved in the recruitment of uninfected cells. The impressive concentration of ICAM at cell sites where most virions are released may account for the acquisition of these membrane proteins by the HIV-1 progeny, and may be important for the cell-mediated spread.

Phenotypic and genotypic alterations characterize patients bearing plasma cell dyscrasias with a high M-component.

As at present only a long-term follow-up can fully determine whether monoclonal gammapathies of undetermined significance (MGUS) will evolve into multiple myeloma (MM), this study attempted to identify other variables connected with the amount of monoclonal component (MC), generally considered as the most reliable marker of malignant evolution. Thirty-four MGUS subjects showing a high MC (> or = 15.0 g/l) but without clinical evidence of MM (MGUS group b), were characterized for their phenotypic and genotypic profile by comparing them either with 40 MM patients or with 24 subjects affected by a benign form of monoclonal gammapathy (MGUS group a) according to the standard criteria. In addition to the usual laboratory markers, the levels of expression of a panel of CD membrane subsets were measured on B and T lymphocytes. Also, the serum level of the p53 mutant protein and the structural alterations of the c-myc oncogene were evaluated. The results show that for MGUS group b patients, an increased M-protein was accompanied by significantly increased levels of peripheral blood CD3+ T cells and oncogenetic aberrations in c-myc. Since a high serum MC level seems to indicate a greater likelihood of malignant transformation for MGUS patients, these findings suggest that this relationship may be a result of the concomitant alterations observed at a phenotypic and genotypic level. Such alterations may be potentially useful as surrogate markers for the transition of benign to malignant (MM) plasma cell dyscrasia.

Thrombocytopenia during immunotherapy with interleukin-2 by constant infusion.

PURPOSE: To elucidate some of the possible mechanisms that lead to interleukin-2 (IL2)-induced thrombocytopenia. PATIENTS AND METHODS: We evaluated retrospectively the effects of immunotherapy with IL2 in 76 patients with disseminated cancer. The lymphokine was administered by constant infusion, daily for 6 days a week for 4 consecutive weeks. RESULTS: A significant decrease in platelet counts was seen after the first 6 days of therapy in all but two patients: 14 patients experienced grade 2 or 3 toxicity, 21 had grade 1 toxicity, and although the decrease in platelet counts could not be graded as toxicity in the remaining 41 patients, there was an average decrease of 32% from baseline platelet counts in 39 (p less than 0.0001). Thrombocytopenia appeared to be secondary to peripheral platelet destruction, since bone marrow biopsy specimens obtained during thrombocytopenia showed hyperplastic megakaryocytopoiesis. IL2 is inactivated by tubular resorption, and severity of thrombocytopenia was strongly correlated with IL2-induced renal dysfunction (p = 0.0004). Additionally, both renal dysfunction and thrombocytopenia were related to total dose of IL2 and were more pronounced in patients with worse baseline renal function and lower baseline platelet counts. The incidence of thrombocytopenia increased with subsequent IL2 therapy: life-threatening thrombocytopenia (less than 25,000/microL) was seen in nine of 57 patients, five of whom required transfusional platelet support. CONCLUSION: On the basis of preliminary observations, we hypothesize that thrombocytopenia induced by IL2 is caused by accelerated clearance of platelets by the reticuloendothelial system.

Differences in the expression and release of DR, BR, and DQ molecules in human cells treated with recombinant interferon gamma: comparison to other interferons.

This study presents a comparative analysis of the effects of different interferons (IFN) on the three recognizable subsets of human HLA class II molecules: DR, BR, and DQ. Both cellular expression and shedding of class II molecules have been determined on three different cell types. The results can be summarized as follows: class II molecules are markedly increased by IFN gamma; IFN beta has a lower enhancing effect, and IFN alpha has only a slight, if any, effect. Kinetically, the action of IFN gamma is prompter and longer lasting than that of IFN beta. DQ expression is much more enhanced by IFN gamma than either DR or BR; IFN beta has the same effect on all three subsets. Parallel changes of the cellular expression and of the shedding of these molecules are observed. A melanoma and a lymphoblastoid cell line and peripheral blood mononuclear cells show qualitatively similar modifications.

Autoreactive response in seronegative homosexual men at high risk for HIV infection.

Peripheral autoreactive T cell response was evaluated by limiting dilution analysis of autologous mixed lymphocyte reaction cultures in 15 subjects at high risk for HIV infection and in 20 normal individuals. The two groups did not show a quantitative difference of peripheral autoreactive T cells, but they showed different kinetics. While controls provided a straight line passing through the origin, the majority of high risk individuals showed a curve with a limited linear portion at high cell concentration, indicating that different mechanisms regulate the autoreactive response in the two groups studied. A follow-up study performed in three high risk and three normal individuals revealed a time-dependent increase of peripheral autoreactive T cells only in high risk subjects. Such increase correlates with the decrease of CD4+ cell number and CD4+/CD8+ cell ratio. Furthermore, the proliferative response of the same three subjects to gp160 peptides suggests a specific cellular reactivity to HIV components. This work has potential importance in understanding some of the early events in HIV infection.

Serum reactivity against an immunoregulatory sequence of HIV p24 in HIV-1-infected subjects.

The aim of this study was to assess the antibody reactivity in HIV-infected subjects against an HIV-1 p24 sequence, p226 (aa226-237), including a seven amino acid epitope showing immunosuppressive activity in vitro and to evaluate the relationship between anti-peptide antibody levels and disease progression. Sera of HIV-infected subjects, at different stages of disease, were compared to control sera in a retrospective evaluation. Recombinant HIV-1 p24 and p24- and control-peptides were used in an enzyme immunoassay as targets for antibodies present in the sera. Antibodies directed against the whole p24 protein and its peptides were found in all the sera studied but at different levels. The anti-p226 reactivity was not significantly different at different clinical stages. Nevertheless, it was inversely correlated to the reactivity directed against the whole protein, that was lower in subjects characterized by low CD4 cell numbers.

Ferritin downregulation in HIV-infected cells.

Levels of serum ferritin are increased in AIDS patients in relation to the progression of the disease. To establish whether or not this in vivo increase could be due to a direct effect of the virus on the infected cells, three HIV-permissive cell lines, the CD4-positive HeLa-T4-6c and C8166 cells and the CD4-negative RD cells, were infected with HIV-1 strains. The expression of ferritin was followed during the course of acute infection, in parallel to other cellular components. Unexpectedly, all three cell lines showed a phase of decrease in their ferritin content after infection by HIV-1, not justified by the modest and late increase of ferritin in the fluids, due to disruption of infected cells. Since ferritin is involved in the control of cell growth and DNA synthesis, its downregulation may be implied both in cell toxicity and DNA abnormalities due to HIV infection.

Recombinant glycoprotein 120 of human immunodeficiency virus is a potent interferon inducer.

Cells infected with human immunodeficiency virus (HIV) induce antiviral activity in peripheral blood mononuclear cells (PBMC) from healthy donors. This activity is neutralized by anti-interferon-alpha antibody and partially destroyed at pH 2. Previous studies with enriched cell populations and monoclonal antibodies suggest that B lymphocytes are the main IFN-producing cells, and that both CD4 and HLA class II antigens are essential for IFN induction. Since the initial event of HIV infection of CD4+ cells is the interaction of the virus coat glycoprotein gp120 with CD4 molecule, we investigated whether gp120 is responsible for IFN induction. Using PBMC and recombinant gp120 obtained from a baculovirus expression system, dose-dependent induction of antiviral activity was observed with titers approaching 10(3) IU/ml. This induction was blocked in the presence of antibody to gp120. The antiviral activity was characterized as IFN-alpha by neutralization with IFN alpha-specific antibody. Preincubation of PBMC with anti-CD4 or the presence of soluble CD4 during incubation inhibited IFN induction, indicating that interaction of gp120 with cell-associated CD4 is responsible for this induction. Neither lymphoproliferation nor interleukin-2 (IL-2) production was observed during IFN induction. However, class G immunoglobulin secretion was enhanced by gp120, indicating that B cells are direct or indirect targets of gp120 stimulation in this experimental system. Since gp120 is shed from HIV-infected cells and occurs in the serum of acquired immunodeficiency syndrome (AIDS) patients, our data suggest that this glycoprotein is responsible for the induction of endogenous IFN and the polyclonal activation of B cells both of which are observed in AIDS patients.

Coordinate induction of interferon alpha and gamma by recombinant HIV-1 glycoprotein 120.

Similarly to HIV-infected cells, recombinant HIV-1 glycoprotein 120 induces acid-labile interferon production in peripheral blood mononuclear cells from healthy donors. Acid lability of this interferon is due to the presence of both IFN-alpha and -gamma molecules. In fact, although not revealed by neutralization of antiviral activity with antibody to IFN-gamma, the presence of IFN-gamma was shown both immunoenzymatically and by detection of specific mRNA in gp120-stimulated cells. The source of IFN-gamma appears to be a T cell present in the CD4-enriched subpopulation. Cultures treated with monoclonal antibodies to the ICAM-1 and LFA-1 adhesion molecules showed an impaired release of both IFN types after gp120 stimulation, suggesting a crucial role of cell-to-cell interactions in the process leading to IFN production. Our data suggest that the HIV envelope glycoprotein could be responsible for the induction of endogenous IFN-alpha and -gamma observed in AIDS patients.

HIV type 1 grown on interferon gamma-treated U937 cells shows selective increase in virion-associated intercellular adhesion molecule 1 and HLA-DR and enhanced infectivity for CD4-negative cells.

Cellular adhesion molecules, such as ICAM-1, -2, and -3; LFA-1; and HLA class I and II are incorporated into HIV-1 virions during budding from infected cells. These virion-associated molecules can be involved in the adsorption to susceptible cells displaying the corresponding counterligands. A number of cytokines have been shown to upregulate the cellular expression of adhesion molecules, such as ICAM-1 and HLA-DR. In this study we investigated the effects of IFN-gamma on the incorporation of ICAM-1, LFA-1, and HLA-DR into mature HIV-1 progeny from chronically infected cells. The ability of such virus progeny to infect either CD4-positive or -negative cells was also investigated. The results indicate that IFN-gamma stimulates the expression of ICAM-1 and of HLA-DR on HIV-1-infected cells, whereas LFA-1 expression is unaffected. The same modifications were also observed on virus progeny, because specific MAbs to ICAM-1 and HLA-DR captured infectious HIV-1 from IFN-treated cells with higher efficiency as compared to virus from control cells, whereas virus binding to anti LFA-1 MAb was unchanged. Moreover, the HIV-1 progeny released from IFN-treated cells showed an increased ability to bind to and to infect CD4-negative cells, whereas the infectivity was basically unchanged for CD4-positive cells. Our results suggest that cytokines, as well as other soluble factors, may expand the host cell range of HIV-1, possibly through modifications of the cell-derived surface molecules on the virions.(ABSTRACT TRUNCATED AT 250 WORDS)

DNA heterogeneity of Staphylococcus aureus strains evaluated by SmaI and SgrAI pulsed-field gel electrophoresis in patients with impetigo.

To our knowledge, no studies have previously been carried out on the heterogeneity and intrafamily colonization of impetigo Staphylococcus aureus strains obtained by powerful discriminating methods such as pulsed-field gel electrophoresis (PFGE). To explore this topic, macrorestriction patterns of S. aureus strains were analyzed after SmaI and SgrAI digestion. The two enzymes provided superimposable results. A total of ninety-seven S. aureus strains was found in the 26 families whose lesions and nasal and pharyngeal samples were examined. There were 39 strains which were different by PFGE, and of these, 24 were found in the lesions. Although 85% of impetigo patients showed nasal colonization and 58% showed pharyngeal colonization, only 54% of the patients had the same PFGE strain in the lesion and in the nose, and 35% in the lesion and the pharynx. In half of the 26 families, at least one member (mother, father, or relative) presented a S. aureus strain identical, by PFGE, to strains isolated in patients' lesions. Nineteen percent of mothers, 15% of fathers, and 19% of the other relatives presented nasal colonization with strains identical to those isolated in the children's lesions. Lesional strains showed higher antimicrobial resistance than nonlesional isolates.

Serum interleukin-10 levels in patients affected with multiple-myeloma - correlation with the monoclonal component and disease progression.

Using a commercially available, competitive ELISA kit based on a polyclonal anti-interleukin-10 antibody, serum interleukin-10 (IL-10) levels were quantified in samples of different groups of patients: 20 healthy controls (CTR), 11 monoclonal gammopathies of uncertain significance (MGUS), 17 multiple myelomas (MM), 10 cancer patients (CANCER), 13 cancer patients + MGUS (MGUS-CA) and 7 MGUS patients after surgical removal of concomitant cancer (MGUS-SRCC). Results show significant differences of both the median levels of IL-10 and the monoclonal component (MC) in CTR, MGUS and MM (patients with increasing concentrations in the mentioned order). The IL-10 levels found in the three groups of cancer patients showed serum levels higher than those observed in the controls. Moreover, the surgical cancer removal was related to an IL-10 decrease. A higly significant correlation between serum IL-10 levels and the corresponding MC was also found in the MM-bearing patients and to a lesser extent, in MGUS patients, indicating that serum IL-10 is parallel to the amount of the activated clone causing the monoclonal gammopathy. Since human myeloma lines, cultured in vitro may release significant amounts of IL-10, the data presented support the hypothesis that serum IL-10, measured in myelomatous patients may, at least in part, derive from the activated clone causing the monoclonal gammopathy.