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The clinical management of patients affected by systemic diseases, including cancer and autoimmune diseases, is generally founded on the evaluation of the only markers related to the single disease rather than the biological immuno-inflammatory response of patients, despite the fundamental role of cytokine network in the pathogenesis of cancer and autoimmunity is well known. Cancer progression has appeared to be associated with a progressive decline in the blood levels of the main antitumor cytokines, including IL-2 and IL-12, in association with an increase in those of inflammatory cytokines, including IL-6, TNF-alpha, and IL-1-beta, and immunosuppressive cytokines, namely TGF-beta and IL-10. On the other hand, the severity of the autoimmune diseases has been proven to be greater in the presence of high blood levels of IL-17, TNF-alpha, IL-6, IL-1-beta, IFN-gamma, and IL-18, in association with low levels of TGF-beta and IL-10. However, because of excessive cost and complexity of analyzing the data regarding the secretion of the single cytokines, the relation between lymphocyte-induced immune activation and monocyte-macrophage-mediated immunosuppression has been recently proven to be expressed by the simple lymphocyte-to-monocyte ratio (LMR). The evidence of low LMR values has appeared to correlate with a poor prognosis in cancer and with a disease control in the autoimmune diseases. Moreover, since the in vivo immunoinflammatory response is physiologically under a neuroendocrine modulation, for the evaluation of patient biological response it would be necessary to investigate the function of at least the two main neuroendocrine structures involved in the neuroendocrine modulation of the immune responses, consisting of the hypothalamic-pituitary-adrenal axis and the pineal gland, since the lack of physiological circadian rhythm of cortisol and pineal hormone melatonin has appeared to be associated with a worse prognosis in the human systemic diseases.

Background: Several clinical studies have clearly demonstrated that the immune status is one a major prognostic factor for the survival time in cancer patients. However the main clinical problem is to identify the most prognostically important index within the great number of immune parameters. Recently the evaluation of regulatory T (T-reg) (CD4CD25) lymphocyte count and function with respect to the T helper (TH) (CD4) number has been shown to represent the main immune parameters capable of representing the functional status of the anticancer immunity in cancer patients. This study evaluated the influence of the four main conventional anticancer therapies (surgery, chemotherapy, radiotherapy, immunotherapy) on the CD4/CD4CD25 ratio., Patients and Methods: The study included 70 patients. The oncological treatments consisted of surgery in 14, chemotherapy in 36, radiotherapy in 12 and immunotherapy (subcutaneous low-dose, S.C.-low, interleukin, IL-2) in 8 patients. The normal value of the CD4/CD4CD25 ratio was greater then 4.0., Results: Surgery induced a significant decline in the CD4/CD4CD25 mean ratio. Radiotherapy also induced also a dramatic significant decrease in the CD4/CD4CD25 ratio, whereas the effect of both chemotherapy and immunotherapy reflected the clinical response to the treatments. The CD4/CD4CD25 mean ratio was significantly enhanced in the patients who obtained control of the neoplastic growth, whereas it diminished in progressing patients., Conclusion: The commonly used anticancer therapies profoundly modify the levels of amounts of T-reg lymphocytes. Because of the fundamental role of T-reg cells in suppressing the anticancer immunity, thus diminishing survival, the monitoring of the CD4/CD4CD25 ratio could constitute an important clinical index during conventional anticancer therapies to predict the prognosis of cancer patients.

A phase-II pilot clinical study was performed to evaluate the effects of low-dose subcutaneous IL-2 with the pineal hormone melatonin (MLT) in AIDS patients with CD4 counts below 200/mm3. The study included 11 patients. IL-2 was given subcutaneously at 3 million IU/ day in the evening for 6 days/week for 3 weeks. MLT was given orally at 40 mg/day in the evening every day, starting 7 days prior to IL-2. The treatment was substantially well tolerated, and in particular no cardiovascular or pulmonary complication occurred. An increase in CD4 cell number greater than 30% occurred in 4/11 (36%) patients, and CD4 cell mean values observed during the study were significantly higher with respect to those found before. In addition, the treatment induced a significant increase in mean number of lymphocytes, eosinophils, T lymphocytes, NK cells, CD25- and DR-positive lymphocytes. Finally, CD4/CD8 mean ratio significantly increased during the study. This preliminary clinical study suggests that the combined neuroimmunotherapy with low-dose subcutaneous IL-2 and MLT may improve the immune status also in AIDS patients with CD4 cell counts below 200/mm3, who generally do not respond to IL-2 alone.

Suppressive events induced by macrophages and TH2 lymphocytes would represent the most important factors responsible for the in vivo reduced efficacy of IL-2 cancer immunotherapy. Previous studies have shown that IL-3 or the pineal hormone MLT may abrogate macrophage-related suppressive events during IL-2 immunotherapy, while TH2-mediated immunosuppression is not neutralized by MLT or IL-3. On the basis of previous experimental data suggesting the inhibitory effect of IL-12 on TH2 activation, this preliminary study has been performed in an attempt to evaluate the influence of IL-12 on TH2 stimulation induced by IL-2 alone or IL-2 plus MLT, by evaluating the release of IL-10, which represents the main suppressive factor produced by TH2 lymphocytes. Pure lymphocyte cultures were incubated for 4 days with IL-2 (100 Cetus U/ml), MLT (100 pg/ml), IL-12 (1 ng/ml), IL-2 plus MLT, IL-2 plus IL-12 or IL-2 plus MLT and IL-12. Mean medium concentrations of IL-10 were measured by Elisa. IL-2 alone significantly stimulated IL-10 secretion with respect to the control medium alone, while no difference was observed with MLT alone or IL-12. IL-2-induced stimulation of IL-10 secretion was not abrogated by a concomitant MLT incubation. On the contrary, IL-12 significantly diminished IL-10 release in response to IL-2, and this inhibitory effect was more pronounced when IL-2 was added in association with both IL-12 and MLT. This preliminary study would suggest that the two most important immunosuppressive events occurring during IL-2 therapy, which are mediated by macrophages and TH2-lymphocytes, may be abrogated by a concomitant administration of MLT and IL-12, respectively. Therefore, the association of IL-12 could further amplify IL-2 efficacy with respect to IL-2 alone or IL-2 plus MLT.

The Sceptor system and the susceptibility of Gram-positive cocci and Gram-negative rods to several antibiotics were presented. The accuracy of methicillin and gentamicin MIC of Staphylococcus aureus was determined. The MIC of methicillin-resistant strain was less than 8 mg/l. Finally the MIC accuracy to 29 Pseudomonas aeruginosa strains was compared with the results obtained by the standard disk diffusion method.

An immunoglobulin M immunosorbent agglutination assay (IgM ISAGA) was tested in 1804 outpatients' samples. The test was negative in all 1091 sera from individual negative in the total indirect immunofluorescence antibody (IFA tot) test and in the passive hemagglutination (HA) test and in 15 sera with inconclusive results in these two tests. The 698 sera positive in the IFA tot and HA tests were also tested with direct IgM enzyme-linked immunosorbent assay (IgM ELISA). The 74 sera positive in the IgM ISAGA and/or IgM ELISA were also tested in the IgM immunofluorescent (IgM IFA) test. These sera belong to 42 individuals, 26 of whom were followed for previous positive results in the IgM ELISA test. 51 samples (68.9%) were found positive in both IgM ISAGA and IgM ELISA tests. Only 13 (25.5%) of these 51 samples were found positive in the IgM IFA. 9 samples (12.2%) were positive only in the IgM ISAGA test. Conversely another 9 samples (12.2%) were positive only in the IgM ELISA test, performed on serum as well as on chromatographic IgM fraction. Finally five samples (6.7%) yielded false positive results in the IgM ELISA test, and three of them were found false positive also in the IgM IFA test. All these five samples in fact were found negative on IgM chromatographic fraction in both tests. In conclusion, IgM ISAGA appears to be more specific than direct IgM ELISA or IgM IFA test for the detection of Toxoplasma gondii IgM. Sensitivity of IgM ISAGA test seems to be as good as in the direct IgM ELISA and better than in the IgM IFA test for the diagnosis of acute acquired toxoplasma infection.

Four different assays for detection of rubella IgG antibodies, two latex agglutination (Rubalex and Rubascan), one hemagglutination inhibition (HAI) and one enzyme-linked immunosorbent assay (ELISA IgG), were used to test 372 human serum samples. All samples were also tested with a rubella ELISA IgM test and all of them were found negative. The results obtained in 358 (96.2%) out of 372 samples tested were identical with all procedures. Inconclusive results were obtained in 14 (3.8%) specimens, in that there were positive and negative results for the different assays. These 14 specimens were sent to the Institute of Virology in Turku (Finland) and tested by using a hemolysis in gel and a non commercially-available ELISA IgG test. Results obtained with the two last tests were considered definitive. The two latex tests were found equal or better than HAI and ELISA IgG for sensitivity and specificity. Furthermore they were also cost effective and more simple to perform.