Evidence for a positive correlation between serum cortisol levels and IL-1beta production by peripheral mononuclear cells in anorexia nervosa.

A hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis has been reported in anorexia nervosa (AN), together with some immunological abnormalities, involving citokine - and particularly Tumor Necrosis-Factor-alpha (TNF-alpha) - production by polymorphonuclear cells. The ability of pro-inflammatory cytokines to activate the HPA axis is well known; however, there are no data demonstrating an interdependence between immunological and endocrine response in AN. To investigate the presence of a correlation between immune response and pituitary-adrenal function, plasma ACTH and serum cortisol concentrations were measured in 13 AN patients and in the same number of controls. TNF-alpha and interleukin (IL)-1beta production by ex-vivo unstimulated and LPS-stimulated peripheral mononuclear cells was also assessed. Circulating cortisol concentrations were higher (p<0.01) in AN (156.7 +/- 45.1 microg/l, mean +/- SD) than in controls (105.9 +/- 25.7 microg/l). Unstimulated IL-1beta release in supernatants of mononuclear cell cultures was slightly but not significantly higher in AN than in controls, while TNF-alpha release was similar in the two groups. A positive correlation was found between IL-1beta concentrations in unstimulated culture supranatants and serum cortisol levels in AN (r=0.782, p=0.002), while in normal subjects there was a trend toward a negative correlation; a slight positive correlation, while not significant, between IL-1beta and plasma ACTH, as well as between TNF-alpha and serum cortisol was also found in AN. These data suggest that the normal relationship between pro-inflammatory cytokines release, particularly IL-1beta, and cortisol secretion is deranged in AN.

Up-modulation of interferon-gamma mediates the enhancement of spontanous cytotoxicity in prolactin-activated natural killer cells.

Prolactin (PRL) has been shown to participate in lymphocyte activation. In particular, the constitutive natural killer (NK) and the lymphokine-activated killer (LAK) cytotoxicity of CD56+ CD16+ cells is increased by its physiological to supraphysiological concentrations. As PRL has been shown to up-regulate the production of interferon-gamma (IFN-gamma) by peripheral blood mononuclear cells, we studied its effect on IFN-gamma production by NK cells as a possible mechanism of autocrine activation of cytotoxicity. Released and intracellular IFN-gamma, as well as IFN-gamma mRNA expression, were increased by pituitary and recombinant human PRL, which stimulated optimal NK and LAK cytotoxicity. Treatment with blocking anti-IFN-gamma monoclonal antibody (mAb) selectively affected PRL-increased killing of K562 targets, demonstrating that PRL-mediated enhancement of spontaneous cytotoxicity depends, at least in part, on up-regulation of IFN-gamma.

Detection of Pneumocystis carinii by DNA amplification in human immunodeficiency virus-positive patients.

The opportunistic pathogen Pneumocystis carinii (PC) is a frequent cause of a life-threatening pneumonia in human immunodeficiency virus (HIV)-infected individuals and in other immunocompromised hosts. Specimens obtained from 128 bronchoalveolar lavage (BAL) fluid samples from 123 HIV-positive patients with pulmonary disease and undergoing a diagnostic bronchoscopy were evaluated to detect this organism. We have developed a rapid DNA extraction procedure for nested polymerase chain reaction (PCR) using two sets of primers (pAZ102-E, pAZ102-H and P1 = 5'-CTAGGATATAGCTGGTTTTC-3' and P2 = 5'-TCGACTATCTAGCTTATCGC-3'). The results were compared using cytological techniques (direct wet mount, Giemsa, toluidine blue O) and related to the clinical follow-up of patients. The nested PCR had a 91% sensitivity and a 93% specificity. The effect of chemoprophylaxis and the evaluation of the follow-up of patients are discussed. Nested PCR may represent an important additional tool, along with current cytological methods, for the detection of P. carinii; however, at present it cannot replace routine microbiological methods more simple and less expensive.

Phytosterol compounds having antiviral efficacy.

The present study is focused on the antiviral action patterns obtained in vitro with synthetic sterolester comprising compositions on virus-bearing host cell-lines. Appropriate cell-lines were infected with HIV-1, human Cytomegalovirus (HCMV) and Herpes simplex virus (HSV). There appears to exist a clear anti-infective efficacy for a selected number of such ester compounds, provided they are formulated into spontaneously dispersible concentrates, which in aqueous dilution engender ultramicro-emulsions having micelles in the lowest nanosize region. A significant protection against HIV-induced cytopathogenic effect was demonstrated employing a methyltetrazolium salt reduction assay on HIV-infected MT4 cells when they were incubated with such concentrates. A similar effect was evidenced with the same concentrates, when preincubating concentrated virus, but not the target cells. Antiviral activity appeared to be remarkable also on HCMV infections in vitro, where a blocking effect on immediate-early antigen expression in fibroblast monolayers could be observed. Similarly, HSV-associated glycoprotein antigen in VERO cells also suggests that virus-cell interaction and/or virus multiplication could have been blocked at a very early point of time. This would be quite different from antiviral action-patterns studied so far and imputed into the current models of explanation. Proper solubilization of the employed phytosterol compounds is essential for achieving the described activity modes. The often recommended liposome formulations would not be well suited for such compounds and such purpose, since after dilution they produce aqueous macro-emulsions, only. Furthermore, liposome formulations tend to coalesce and exhibit Marangoni effects.

Serum cytokine profiles in acute primary HIV-1 infection and in infectious mononucleosis.

Serum cytokine profiles, T-cell subsets, and general parameters of immune activation were evaluated in 15 patients with acute primary HIV-1 infection, and compared with those obtained from 18 patients with acute primary Epstein-Barr virus (EBV) infection and from 18 control subjects in order to elucidate possible defects of immune response to HIV in early phases of virus-host interaction. Mean CD4+ cell count, serum concentrations of interleukin (IL)-2, IL-4, soluble IL-2 receptor (sIL-2R), tumor necrosis factor (TNF)-alpha, 5'-neopterin, and beta 2-microglobulin were significantly lower in acute HIV-1 infection than in EBV infection. Both acute HIV-1 and EBV infections were characterized by significantly higher mean CD8+ cell count and soluble CD8 antigen (sCD8) levels compared to control subjects, while acute HIV-1 infection was accompanied by the highest interferon (IFN)-gamma serum concentrations. In primary HIV-1 infection, significant impairment of CD4+- mediated T-helper function may lead to viral escape and persistence of infection despite an early and vigorous CD8+ T-lymphocyte activation.

Clinical and Diagnostic Aspects of Travelers' Diarrhea due to Cyclospora Organisms.

Cyclospora sp, a recently described protozoan, is associated with prolonged self-limiting and relapsing diarrhea. The species has a worldwide distribution and a high prevalence in tropical countries. Some reports suggest that the agent is a common cause of travelers' diarrhea.

Clinical and microscopical features of small-intestinal microsporidiosis in patients with AIDS.

Intestinal microsporidiosis by Enterocytozoon bieneusi is an increasingly recognized infection in AIDS patients. We report eight cases of microsporidiosis. All patients were severely immunodepressed. Clinical features were highly variable. Patients were followed up for a mean period of 7.8 months. All patients had persistent infection during the follow-up and spore excretion remained constant. Two patients became asymptomatic during the follow-up. None of the patients presented clinical and echographic signs of biliary involvement. Treatment with albendazole, metronidazole or paromomycin failed to produce a durable clinical response or to eradicate the organism. Cases were identified by stool examination and additionally investigated with light and electron microscopy. It was found that light microscopy was a sensitive method, while electron microscopy was less sensitive but allowed the definition of the infecting species. The modified trichrome stain was a satisfactory method for diagnosis on fecal smears. The calcofluor stain and the combination of DAPI with calcofluor was a rapid and simple staining method for screening.

Altered adrenocorticotropin and cortisol response to corticotropin-releasing hormone in HIV-1 infection.

Alterations of the hypothalamic-pituitary-adrenal (HPA) axis are common in HIV infection. To characterize further the site of these derangements and their possible causes, eight male drug addicts with symptomatic HIV infection (stage IV C2) underwent the following investigations: repeated baseline determinations of cortisol, adrenocorticotropin (ACTH), interleukin 1 beta (IL-1 beta), IL-6 and interferon alpha (IFN-alpha); and ovine corticotropin-releasing hormone (CRH) test (100 micrograms IV) for ACTH and cortisol determinations. Baseline cortisol levels were either normal or elevated in all patients. A significant linear correlation was found between baseline levels of cortisol and both IL-6 (r = 0.955; p < 0.001) and IL-1 beta (r = 0.863; p < 0.005), but not between cortisol and ACTH or between ACTH and circulating cytokines. Both ACTH and cortisol responses to CRH were nearly absent in six out of eight patients, and delayed in the others. The areas under the curves of both ACTH and cortisol after CRH were significantly lower in HIV patients than in a group of eight healthy control subjects (p = 0.0157 for ACTH and p = 0.046 for cortisol). Out data suggest the possibility of an inappropriate stimulation of the HPA axis in symptomatic HIV infection by HIV-induced release of cytokines, with a blunted pituitary and adrenal response to CRH.

Interaction of human plasma fibronectin with viral proteins of human immunodeficiency virus.

Fibronectin (FN) is present in soluble and matrix forms in various body fluids and tissues, and has been shown to bind to several pathogens, including viruses. The interaction of FN with viral proteins of human immunodeficiency virus (HIV-1) was investigated by immunofluorescence technique using a cell line chronically infected with HIV-1 (H9-V). The results of this study showed that FN binds to HIV-1 infected cells, especially at FN concentration of 5 micrograms/ml. In addition, FN-pentapeptide has shown the ability to bind to HIV-1 infected cells. On the other hand, preincubation with antibodies against FN abolished the binding of FN to HIV-1 infected cells. Finally, FN has shown to bind to HIV-1 glycoproteins, including gp41 and gp120. In contrast, no binding to HIV-1 core proteins, including p15 and p24, was noted. We suggest that FN, in binding HIV-1 particles, may reduce viremia and thus may be involved in the clearance of viral proteins from the cells.

Cytokine network and acute primary HIV-1 infection.

OBJECTIVE: To investigate the relationship between cytokine serum levels, peripheral blood lymphocyte subsets and clinical picture in acute primary HIV-1 infection. PATIENTS AND METHODS: Absolute number/microliters total lymphocytes, CD4+, CD8+ and natural killer (NK) cells, as well as serum levels of soluble CD8 receptor, interleukin (IL)-1 beta, IL-2, IL-4, IL-6, tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, beta 2-microglobulin and 5'-neopterin were determined in 15 patients with acute primary HIV-1 infection, 16 asymptomatic HIV-1-seropositive individuals and 18 HIV-1-seronegative individuals at risk for HIV-1 infection. RESULTS: Acute primary HIV-1 infection was characterized by significant CD4+ lymphocytopenia with low IL-2 serum concentrations, and by high absolute number of circulating CD8+ and NK cells, with elevated serum levels of soluble CD8 receptor, IL-1 beta, IFN-gamma and 5'-neopterin. Follow-up of acute seroconverters showed a significant decrease in NK cell counts and IL-1 beta levels, with an increase of IL-6. CONCLUSIONS: In acute primary HIV-1 infection, significant alteration of cytokine release, possibly induced by viral antigens, could be responsible for both clinical picture and activation of cytotoxic cells through abnormal mechanisms.

Alveolar immune mediators in HIV-related pneumonia. Different role of IL-2 and IL-1 in inducing lung damage.

In order to elucidate the role played by alveolar cytokines in the pathogenesis of HIV-related lung damage, levels of interleukin (IL) 1 beta, IL-2, IL-6, tumor necrosis factor (TNF)-alpha, and interferon (Ifn) were assessed on supernatant of bronchoalveolar lavage fluid from 30 consecutive HIV-1 seropositive (HIVAb+) patients with clinical and radiologic evidence of pneumonia, from 20 HIV- seronegative (HIVAb-) patients with pulmonary sarcoidosis, and from 10 HIVAb- healthy control subjects. Cytokine levels were expressed as picogram (IL-1, TNF), nanogram (IL-6), and international unit (IL-2, Ifn) per milligram of albumin per deciliter. Total and differential cell counts, cytofluorimetric enumeration of CD3+, CD3+/DR+, CD4+, CD8+, and CD8+/CD16+ cells, as well as microbiologic investigations for opportunistic agents were performed on lavage pellets. HIV-related pneumonia was characterized by higher mean alveolar level of IL-2 (12 +/- 5 IU), and by more elevated mean counts of T cells (109 +/- 16), activated T cells (60 +/- 12), and CD8+ cells (90 +/- 13)/microliters if compared with both active sarcoidosis and control subjects, where respective values of 0.2 +/- 0.1 and 0.3 +/- 0.2 IU IL-2/mgAlb/dl, of 52 +/- 11 and 7 +/- 2 T cells, of 20 +/- 5 and 1.2 +/- 0.3 activated T cells, and of 11 +/- 2 and 3 +/- 0.6 CD8+ cells per microliter were found. HIV-infected patients with opportunistic lung infections (OIs) showed the highest mean IL-2 level (21 +/- 4 IU), and higher counts of both CD8+ (117 +/- 20) and CD8+/CD16+ (36 +/- 7) cells per microliter if compared with patients without evidence of OIs (respectively, 62 +/- 13 CD8+ and 18 +/- 3 CD8+/CD16+ cells per microliter). By contrast, extremely high IL-1 levels (1,463 +/- 760 pg), and IL-2 levels similar to control subjects (3.4 +/- 1.2 IU), were found in the absence of OIs. Different mechanisms depending respectively on IL-2-mediated cytotoxic cell recruitment and activation, or IL-1-mediated tissue injury may account for HIV-related lung damage, depending on the presence or absence of opportunistic agents.

Meningoencephalitis caused by Toxoplasma gondii diagnosed by isolation from cerebrospinal fluid in an HIV-positive patient.

Toxoplasmosis of the central nervous system is a frequent opportunistic infection in AIDS patients, usually presenting as a mass brain lesion detected by computerized axial tomography scanning or magnetic resonance imaging. A case of diffuse meningoencephalitis with no radiological evidence of brain lesions is described. Diagnosis was made by culturing cerebrospinal fluid (CSF) on THP1 cells where tachyzoites of Toxoplasma gondii were demonstrated after 8 days of incubation by both direct observation and immunofluorescence. CSF examination with culture should be considered in AIDS patients with neurological signs and symptoms but without radiological evidence of cerebral lesions.

Spontaneous release of interferon as a predictor of clinical evolution in HIV-positive subjects.

In order to establish a correlation with disease progression we prospectively evaluated ten clinical and immunologic parameters in 102 consecutive HIV-positive subjects. The eight immunologic variables were: in vitro spontaneous interferon release by peripheral blood monocytic cells, alpha- and gamma-interferon production induced by Newcastle Disease Virus and PHA, Multitest Mérieux score, PHA- and CON-A-induced lymphocyte transformation, absolute number of CD4+ cells and CD4/CD8 ratio, respectively. The two baseline clinical variables were risk factor and disease presentation. Generalized Wilcoxon analysis indicated a significant correlation of one clinical (disease presentation at entry) and three immunologic variables (spontaneous interferon release, CD4+ cell number, Multitest Mérieux) with disease progression. Baseline spontaneous release of alpha, acid-labile interferon showed the best correlation with disease progression, and appeared to be significantly associated with CD4+ cell loss. Spontaneous release of acid-labile alpha interferon by mononuclear cells in vitro could be highly predictive of disease evolution in HIV-Ab positive, AIDS-free subjects.

Effects of long-term zidovudine treatment on cell-mediated immune response and lymphokine production.

The effects of long-term zidovudine treatment on functional parameters of cell-mediated immunity were investigated in 15 symptomatic HIV-antibody-positive patients with clinical evidence of opportunistic infections. Mononuclear leukocytes were obtained before administering the drug, and after 3 and 6 months of treatment. The cells were stimulated with lectins in order to assess variations of mitogen-induced lymphocyte proliferation and production of gamma-interferon (IFN) and interleukin-2 (IL-2), and with Newcastle disease virus (NDV) to assess variations of alpha-IFN production. Mean proliferative responses to PHA and Con-A did not show any significant change between baseline, 3 months, and 6 months values (25,158 +/- 11,763, 24,662 +/- 8,955, and 34,924 +/- 16,283 D-cpm, respectively, for PHA, p greater than 0.05; and 5,470 +/- 1,890, 4,953 +/- 2,518, and 4,539 +/- 3,286 D-cpm, respectively, for Con-A, p less than 0.05). Mean response to PWM (9,707 +/- 4,429 D-cpm at entry) increased significantly after 3 months, but returned to baseline values at 6 months (17,039 +/- 5,123 and 10,314 +/- 3,855 D-cpm, respectively, p = 0.016). IL-2 production (5.51 +/- 4.0 I.U. at entry) rose particularly at 3 months and persisted at the same levels at 6 months (9.6 +/- 4.8 and 9.5 +/- 6 I.U., respectively, p less than 0.05). By contrast, a moderate but significant decrease in gamma- and alpha-IFN production was observed (65 +/- 2.2, 35.1 +/- 1.6, and 22 +/- 2 I.U., respectively, for gamma-IFN, p less than 0.05; 60 +/- 3, 64 +/- 2.6, and 12 +/- 1.5 I.U., respectively for alpha-IFN, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of ofloxacin on cell-mediated immune response and lymphokine production.

The effects of orally administered ofloxacin on functional parameters of cell-mediated immunity were investigated in 15 patients with respiratory or urinary tract infections. Mononuclear leucocytes were obtained before administering the drug, 1 h after the first dose, and five days later. The cells were stimulated with lectins, tetanus toxoid and Newcastle Disease virus in order to assess mitogen- and antigen-induced lymphocyte proliferation and production of gamma-interferon, interleukin-2 and alpha-interferon. An increase in proliferative response to pokeweed mitogen and a slight but significant decrease in alpha-interferon production were observed, while other parameters remained unaffected by treatment.

[Biological properties of an alcoholic extract of Aspergillus terreus]. / Proprietà biologiche di un estratto alcoolico di Aspergillus terreus.

The following effects of Aspergillus terreus alcoholic extract are investigated: antiblastic and antiviral effect, and the ability of modification of interferon production and lymphocyte blastic transformation. In particular the extract showed more evident antiproliferative effect on transformed or EBV immortalised cells and lower effect on normal cells. Moreover doses from 12.5 +/- micrograms/ml depressed significatively the in vitro interferon production and lymphocyte blastic transformation induced by PHA on human cells. Mengo and Semliki Forest viruses replication was reduced, although temporarily, in the presence of 50 +/- micrograms/ml of the extract. Eventually the study of some fractions separated on chromatographic column demonstrated the contemporary presence of fractions able to inhibit or stimulate K-562 erythroleukemic cells replication.

Effect of erythromycin on the immune response and interferon production.

The influence of erythromycin on some aspects of humoral and cell-mediated immunity has been examined employing human as well as murine models, both in vivo and in vitro. No significant differences in antibody synthesis, alpha- and gamma-interferon yield, cutaneous delayed hypersensitivity or lymphocyte blastogenic response to mitogens have been detected between erythromycin-treated subjects and controls. Similarly, in vitro tests on interferon production and blastogenic response to mitogens showed no significant differences when performed with and without erythromycin. Therefore, in contrast with many other antibacterial drugs, erythromycin seems to be devoid of any adverse effects on the immune system.

[Viral infections and diabetic disease]. / Infezioni virali e malattia diabetica.

Authors report recent data from medical literature on virus infections effect on diabetogenesis. Particularly they examine Rubella, Mumps, Coxsackie, Encephalomyocarditis and Herpesvirus infections which seem be more active on this process. Finally authors analyze the possible biological mechanism of viral diabetogenesis.