Search

Zusammenfassung Die zellvermittelte Immunitat bei der mannlichen Fertilitat Der Leukocyten-Migrations-Hemmtest auf Agarose wurde bei 210 Mannern verschiedener Provenienz durchgefuhrt (Oligozoospermie als Folge von einem primaren Hodenschaden; Oligozoospermie als Folge von primaren, nicht testikularen Schaden; idiopathische Oligozoospermie). Es kann gezeigt werden, das in keinem Falle von normaler Fertilitat und von Mannern mit Oligozoospermie (nachgewiesene Fertilitat) ein positiver Test-Befund zu erheben war. Bei einseitigem und beiderseitigem Hodenhochstand fand sich in 48% ein positiver Test. Bei Hodengewebsveranderungen war der Test in 53% positiv, bei Storungen der Funktion des Keimepithels in 81%, bei Infektionen des Genitaltraktes in 39%, bei einseitiger Varikocele in 31% und bei idiopathischer Oligozoospermie in 33%. Es hat demnach den Anschein, das die zellvermittelte Immunitat involviert ist bei der Pathogenese der Consequenzen der genannten Storungen.

Recent data have shown the presence of immunological alterations in adult patients suffering from obsessive-compulsive disorder (OCD). The objective of this study was to examine the possible effects of 12 months of treatment with different serotonergic drugs, such as clomipramine and selective serotonin reuptake inhibitors (SSRIs) on peripheral immunological cells of 18 OCD patients. Both the absolute number and percent of CD4+, CD8+, CD3+, CD19+and CD56+cells were measured in peripheral blood before and after treatment by means of a Facstar Flow Sorter apparatus. At baseline, all patients showed a significant increase of CD8+and decrease of CD4+lymphocytes when compared with a similar group of healthy control subjects; after the treatment, CD8+and CD4+cells, respectively, decreased and increased significantly, and the CD4+/CD8+ratio increased, when compared with baseline values, in parallel with the clinical improvement. These data suggest that the alterations of immune cells reported in patients with OCD at baseline may be reverted by treatment with SRIs and should be considered a state-dependent marker, perhaps related to a condition of stress.

The presence of Gc (vitamin D binding protein) has been consistently demonstrated on the membrane of B lymphocytes. This protein appears to be spatially associated with surface immunoglobulins. The origin of this surface protein has not yet been determined and the purpose of the present paper was to investigate if Gc may bind to human lymphocytes after immunoglobulin (Ig) capping. For this purpose the presence of Gc on B lymphocytes was examined by three different approaches. First, when cells were examined by immunofluorescence and quantified by flow cytometry, membrane Ig capping was followed by a dramatic decrease in positivity for Gc when compared to native cells. In addition, incubation of capped cells with purified Gc was followed by a significant increase in fluorescence, indicating that this protein had been able to bind again. Second, analysis of solubilized lymphocytes by Western blotting showed that native lymphocytes and capped cells incubated with purified Gc contained a large quantity of a 56kDa protein which was immunoreactive with anti Gc antibodies. This protein band was much weaker on blots from capped cells not treated with Gc. Third, radiobinding assays indicated that, following capping, cells were able to bind Gc in saturable fashion. These results suggest that membrane Gc could play a role in the entry of vitamin D metabolites into lymphocytes.

With the conventional methods used in current practice, the results of human granulocyte motility assays are expressed by a numerical value that defines the leading front of cells into micropore filters. The authors have developed a fast procedure which allows the complete curve of polymorphonuclear leukocyte (PMN) migration to be obtained, from the initial plane of a filter to the maximum propagation depth. In this way, more information is given than that expressed by a single parameter. The paper describes the procedure used to determine the normality curves both for random and stimulated migration, according to a simple model which defines the PMN propagation in micropore filters. A normality band that defines variations due to the stochastic process is also reported. © 1995 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.

Several studies have shown that sera from patients with Hodgkin's disease contain factors capable of inhibiting polymorphonuclear functions, among them chemotaxis. In the present study, we investigated whether these sera, which were able to inhibit PMN chemotaxis in the agarose test, were also able to affect the 3H-NECA binding to PMN membrane obtained from healthy donors. Control experiments were carried out using PMN incubated with a pool of sera from healthy volunteers. No significant difference was found in the maximum number of binding sites; on the contrary, the equilibrium dissociation constant was significantly increased in the membrane preparation of PMN incubated with pathological serum.

Vitamin D binding protein (Gc) is present on the surface of several blood cells and may interfere with the activity of 1,25(OH)2D3. It has previously been reported that Gc may bind to the U-937 line which is known to differentiate upon exposition to 1,25(OH)2D3. In the present paper, we evaluate the expression of Gc on the surface of U-937 and HL-60 lines. Both cell lines did not express Gc on their surface but U-937 cells were able to bind human purified Gc added to the medium whereas HL-60 were not. After culturing with 1,25(OH)2D3, HL-60 became able to bind Gc. This property seems to be related to the monocytic differentiation induced by 1,25(OH)2D3. Conversely, when present together, 1,25(OH)2D3 reduces binding on U-937.

The identification of Gc (vitamin D binding protein) with the anionic polypeptide cochemotaxin has recently been reported. In this paper we investigate its dose dependent cochemotactic activity and report the inhibition of Gc enhanced chemotaxis by vitamin D3. These results further support the role of immunomodulating hormone played by vitamin D.

It has been shown that low-frequency electromagnetic fields may inhibit or enhance the lymphocyte response to many mitogens. How this happens is not known, and many reports suggest that alterations of surface receptors may be involved. Results presented here indicate that cellular activation is inhibited by a high-intensity EMF after the early phases of signal transduction, but it may be enhanced by a low-intensity EMF.

Controversies exist regarding the impact of psychological stress on the functioning of the immune system in humans. The aim of the present study, therefore, was to evaluate whether the condition of a pre-exam stress may or not modify resting lymphocyte subsets, as well as blood pressure and heart rate. About 22 medical residents of both sexes not suffering from any medical or psychiatric disorder were included in the study. Anxiety levels were measured by means of the Hamilton rating scale for anxiety (HRSA) and anxiety traits by means of the panic-agoraphobic spectrum self-report (PAS-SR) version and the obsessive-compulsive spectrum self-report (OBS-SR) version. The results showed that systolic blood pressure and heart rate increased significantly just before sitting an examination (t(1)) in all subjects, as compared with a calm situation (t(2)), in parallel with the increase in the HRSA total score, while no significant difference was observed in lymphocyte subsets at the two assessment times. However, men had a higher number of CD4+ cells than women at t(1) and t(2), while women showed a higher heart rate at t(1). In addition, significant correlations between CD4+ lymphocyte count and heart rate at t(1) or HRSA at t(2) were detected. These findings indicate that the acute stress determined by sitting for examination provokes changes in autonomic nervous system parameters, such as blood pressure and heart rate, as well as in the subjective feeling of anxiety, as shown by the increased HRSA total scores, which were not paralleled by modifications of lymphocyte subsets. However, individual differences, related to both sex and personality traits yet to be identified, seem to have an impact in shaping the stress response.

Allergic rhinitis is regulated by the local production and release of several cytokines. The levels of Th2 cytokines IL-4, IL-6, IL-10 and the Th1 cytokine IFN-gamma were studied in nasal mucus from 30 subjects with allergic rhinitis and 45 non-atopic healthy controls. In this study a sampling technique for collecting nasal mucus, well tolerated by the subjects and with a minimal stimulation of the mucosa, was performed. The cytokine concentrations in nasal mucus samples were detected and quantitated by a new paramagnetic particle-based immunofluorescent assay system more sensitive than the conventional ELISA techniques. The new technique showed reliable values of the measured parameters. The nasal mucus from allergic patients contained significantly higher concentrations of IL-4 (25.5 +/- 3.6 pg/ml; P0.001) and IL-10 (1300 +/- 190 pg/ml; P0.05) compared to the nasal mucus from control subjects (15.2 +/- 2.3 and 532 +/- 28 pg/ml, respectively, for IL-4 and IL-10). No significant modification in IFN-gamma levels of allergic patients was found when compared to control group (respectively, 19.9 +/- 3.3 vs. 25.7 +/- 5.1 pg/ml; P0.05). Moreover, the allergic patients showed lower levels of IL-6 concentrations in the nasal mucus compared to control subjects (64.8 +/- 9.1 vs. 129.0 +/- 18.1 pg/ml; P = 0.0099). These data can be interpreted by the hypothesis that in response to environmental allergens there is a preferential Th2 polarity by activated CD4+ T cells and that the cytokines IL-6 and IL-10 have, respectively, an important anti-inflammatory and counterregulatory action in the pathogenesis of allergic rhinitis.

Background: Some recent findings suggest the involvement of autoimmune mechanisms in childhood onset of obsessive-compulsive disorder (OCD), on the basis of a parallel drawn with Sydenham's chorea, a manifestation of rheumatic fever. A monoclonal antibody called D8/D17 characterizing a B-lymphocyte antigen, present in almost all patients with rheumatic fever, has been found also in children affected by OCD, Tourette syndrome, and chronic tics to a greater degree than in healthy control subjects. The few observations of disturbances of some immunologic parameters in adult OCD patients, prompted the authors to investigate and compare subsets of peripheral immunological cells for differences in adult patients with OCD and healthy control subjects. Methods: Twenty patients suffering from OCD, with no comorbidity for other psychiatric disorders, were compared with a similar group of healthy control subjects. The immune subsets were measured by flow cytometry. Results: The CD8 + lymphocytes were significantly increased and CD4 + lymphocytes significantly decreased in OCD patients, while the other cells did not differ between the two groups. No correlation was found between immunologic and clinical parameters. Conclusions: These data indicate that patients with adult OCD showed increased CD8 + , i.e., suppressor T lymphocytes, and decreased CD4 + , which identify helper T lymphocytes, as compared with a similar group of healthy control subjects. The findings appear peculiar to patients with OCD and are suggestive of an immunologic imbalance, which might be related to the stress deriving from the frustrating situation determined by the disorder itself.

We used low-dose anti-D immunoglobulins for home treatment of Rh+ adult patients with chronic immune thrombocytopenic purpura (ITP). After informed consent, 15 unselected outpatients (ten males and five females, aged 22 to 72), affected by chronic ITP with negative HIV test, were given intramuscular injection of 900-1500 micrograms of anti-Rh0 (D) IgG over 3 days every month for 2 or 3 consecutive months. Platelet count (mean +/- SD) significantly increased from basal value of 17,000 +/- 9,000/microL to 72,000 +/- 55,000/microL at the end of treatment. Eight patients achieved a rise in platelet count above 50,000/microL (five above 100,000/microL) and two of them maintained the increase longer than 6 months without further anti-D administration. Three patients responsive to the first cycle responded to further treatment with substantially identical results. Seven patients had no response. Four of them had not responded to previous glucocorticoid and intravenous IgG therapy. Direct antiglobulin test became strongly positive in all patients and mean serum haptoglobin decreased from a basal value of 118 +/- 59 to 61 +/- 43 mg/dL; nevertheless no clinically overt hemolysis was observed in any patient, there was no rise of serum indirect bilirubin and hemoglobin level was unchanged (mean +/- SD basal level 13.6 +/- 2.2 g/dL; after anti-D 13.9 +/- 1.2 g/dL). No hematoma developed at the injection site, and no other side effects occurred. Our results show that anti-D therapy is effective in the majority of patients well tolerated, and feasible as home treatment: thus it can be recommended as a cheap and safe alternative treatment in ITP Rh+ patients.

A flow cytometric method to detect and study human eosinophils in whole blood was established. Normal subjects and patients with various types of eosinophilia (hypereosinophilic syndromes, allergic diseases, dermatitis, Hodgkin's Disease, parasitosis) were studied. Whole blood samples were treated for 10 minutes at room temperature with a commercially available reagent (FACS Lysing Solution, Becton Dickinson) which acts both as a fixative and as a lysing agent. Eosinophils were identified as a granulocytic subpopulation with higher SSC and FSC properties. This cell population was characterized by evident autofluorescence and hypodiploid DNA features after propidium iodide staining. The purity of the eosinophil population sorted after electronic gating was close to 100%. A very significant correlation between eosinophil counting by our whole blood method and other two assays, namely routine automatic counting by the H*3 Bayer System and eosinophil detection by depolarized SSC, was obtained. The phagocytic properties of eosinophils were also studied by means of a commercially available diagnostic kit, thus demonstrating that our method is also suitable for the study of those granulocytic functions which can be evaluated by flow cytometry.

In this study F-actin polymerization in neutrophils from 21 patients affected by myelodysplastic syndromes (MDS) was evaluated by means of a flow cytometric assay. Neutrophils were stimulated with formyl-methionyl-leucyl-phenylanaline (fMLP; 10(-8) M final concentration) for 15, 30, 60 and 120 sec, and F-actin content was determined using fluorescein-isothiocyanate phallacidin as a specific probe. Eight normal subjects were studied as controls. We found that F-actin polymerization was defective in ten patients, with very impaired values after 60 and 120 sec of stimulation with fMLP. The remaining 11 patients showed a prevalent neutrophil population with normal F-actin polymerization and neutrophil sub-populations with either defective or undetectable F-actin polymerization. In the first group, patients with very poor prognosis (refractory anemia with excess blasts, refractory anemia with excess blasts in leukemic transformation, trisomy 8, multiple karyotypic abnormalities) were present, although patients with aberrations of karyotype were present in the second group. It is possible that defects in neutrophil F-actin polymerization may be responsible for neutrophil dysfunction, which has frequently been observed in MDS.

The motility of circulating neutrophils from seven patients affected by intermediate and high-grade non-Hodgkin's lymphoma was investigated before and after rhG-CSF administration (5 micrograms/kg/d for 5 d subcutaneously) in the course of chemotherapy. Random motility and bacterial lipopolysaccharide-induced chemotaxis were studied by the micropore filter technique in a Boyden chamber. These functions were evaluated by a very sensitive technique, based on a computer-assisted image processing system, capable of giving several parameters about the kinetics of cell migration. Along with a significant increase in neutrophil number, a significant decrease both in random and stimulated motility was found. The kinetics of cell migration showed that the cells maintained the typical gaussian pattern of random motility. On the contrary, neutrophils were found to have lost the typical stimulated migration peak. These findings are consistent with a rhG-CSF-induced impairment of the directional movement, rather than of the ability of moving at random. These effects were found in patients who, in the same experimental conditions, had displayed an enhanced phagocytosis and phagocytosis-associated chemiluminescence along with an enhanced CD32 expression, not due to an aspecific cell manipulation. Two hypotheses may be taken into account: (i) an increased adhesiveness due to a direct or an indirect activity of the cytokine; (ii) an abnormality in the cytoskeleton maturation and/or rearrangement during the accelerated bone marrow transit of myeloid cells. These findings emphasize that rh-GCSF administration can modulate several functions which play an important role in host defence, and suggest the utility of carrying out further studies to investigate the optimum dosage both to correct neutrophil number and preserve neutrophil functional activities.

In clinical practice, reports have been made on immunosuppression after surgical excision of primary tumor or at relapse. However, the relationship between undefined or overt metastases and the host immune system has not been sufficiently examined over a prolonged period. These aspects were investigated in 160 breast cancer patients followed up post-operatively with serial controls over a long period. One hundred and thirty-four cases (91 node negative, (N-), 43 node positive (N+)) were disease-free and 26 relapsed. In all patients, serum T cell populations, serum B lymphocytes and skin reaction of delayed hypersensitivity (SRDH) were serially determined for 39 +/- 12 months (m +/- SD). The reference values for these parameters were assessed as follows: T populations were evaluated in 24 healthy donors and SRDH in 95 healthy females. In non-relapsed patients, constant CD8+ T cell decrease and T4/T8 ratio increase were observed; the T4/T8 ratio was significantly higher (ranging from P0.05 - P0.001) than in the control group. The mean values of NK cells and B lymphocytes, the former parameter being highly significant (P0.001), were higher than in controls. In the 26 metastatic patients, the T4/T8 ratio from 20 months before to 30 months after the first sign of relapse decreased from 3.2 to about 1 (r = -0.256, P0.05) and from 30 to 92 months after relapse progressively increased to 2. Similarly, in the former subinterval a progressive decrease in the number of positive antigens and score was found (from 2.4 to 0 and from 10 to zero respectively). A significant inverse correlation between these two parameters and observation time occurred (P0.01 and P0.001 respectively). From 30 to 86 months after relapse, a progressive increase in the number of positive antigens and scores up to 2 and 12 were observed. A significant direct correlation (P0.05) was noted. In conclusion, these data indicate significant changes in T populations during the disease-free interval in breast cancer patients. The decrease in circulating CD8+ T cells is compatible with the hypothesis of CD8+ T cell localization at the site of the micrometastases. The increase in circulating B lymphocytes and NK cells suggests activation of aspecific humoral immunity and NK function. In addition, they show that progressive deficiency in cell-mediated immunity appears many months before and that recovery continues for a long time after overt metastatic disease.

Isolated resting platelets are able to limit neutrophil activation and then can control the tissue-damaging potential of activated neutrophils. In the present study, platelet-neutrophil interactions have been evaluated in 10 uremic patients; the blood samples have been collected before the hemodialysis session. Twelve normal subjects served as controls. Platelets and neutrophils have been isolated and recombined in an autologous ex vivo system. Anion superoxide production and chemiluminescence (which is related to hypochlorous acid production) have been evaluated after stimulation with N-formyl-methionyl-leucyl-phenylalanine. Coincubation of platelets from normal subjects with autologous neutrophils led to a dose-dependent inhibition of both superoxide anion generation induced by N-formyl-methionyl-leucyl-phenylalanine and chemiluminescence. Instead, platelets from uremic patients have not affected superoxide anion production by autologous neutrophils. The chemiluminescence was reduced by coincubation with autologous platelets only at the highest platelet-neutrophil ratio (100:1). In conclusion, the modulation exerted by platelets towards neutrophil activation can be impaired in chronic uremia. Therefore, the tissue-damaging potential of circulating neutrophils, due to toxicity by superoxide anion and hypochlorous acid, may be increased.

Essential thrombocythaemia (ET) is frequently associated with neutrophil and platelet dysfunction, and with increased incidence of vascular complications (thrombosis, haemorrhage). Several interactions between platelets and neutrophils have been reported, and the reciprocal actions between these cells may have an important role both in thromboregulation and in diseases such as those caused by uncontrolled neutrophil activation. In the current paper the authors studied 15 patients affected by ET and 10 normal subjects as controls. Circulating neutrophils and platelets were purified and were recombined in constant ratios (50 : 1, 100 : 1 and 200 : 1) and the individual platelet to neutrophil ratio. Superoxide anion (O 2 - ) generation and luminol-enhanced chemiluminescence (CL) were studied after neutrophil stimulation with fMLP. In normal subjects both O 2 - generation and CL were inhibited by autologous platelets in a dose-dependent manner. In ET patients, on the contrary, platelet-dependent inhibition of O 2 - generation did not occur, while a dose-dependent inhibition of CL was observed. Two groups of ET patients were found : patients with neutrophil O 2 - generation and CL within the normal range, and patients with significantly reduced neutrophil respiratory burst. However, no differences were found between these two groups of patients in terms of platelet effects towards fMLP-stimulated neutrophils. Therefore, platelets from ET patients were not able to exert the homeostatic control towards neutrophil O 2 - generation shown by platelets from normal subjects, and this phenomenon may have a role in the clinical setting. In fact, O 2 - has been shown to be a very strong direct platelet activator, is able to inactivate nitric oxide (which is a powerful inhibitor of platelet aggregation and adhesion to endothelium), and is directly involved in neutrophil-mediated tissue damage.

Luminol-enhanced whole blood chemiluminescence of human neutrophils was studied using opsonized zymosan as a stimulus. Heparinized blood (0.5 microliters) was used, and the chemiluminescence signals were recorded by a very sensitive, automated, and computer-assisted luminometer (LB 950, Berthold, Wildbad, Germany). The following parameters were provided: integral values over the total measuring time, peak values, the time to reach maximum value, and the time to reach half maximum value. Normal subjects, neutropenic patients, subjects with total or partial myeloperoxidase deficiency, patients with recurrent infections, phagocytic defects, thrombocythemic patients and those with non-Hodgkin's lymphomas undergoing therapy with recombinant human granulocyte colony-stimulating factor were studied. The integral response of chemiluminescence and the time of reach half maximal value were useful indicators of chemiluminescence defects; the assay, was able to detect chemiluminescence responses in neutropenic subjects with neutrophil levels as low as 0.6 x 10(9)/l; differences between cellular and plasma defects could be identified; the quenching effect exerted by erythrocytes was negligible.

FcRIII (CD16) expression an neutrophils from 17 patients with chronic myeloid leukemia (CML) was studied by flow cytometry using monoclonal antibodies. A variable proportion of CD16-negative neutrophils were found both in CML patients in chronic phase (3 out of 8 patients) and in CML patients in hematological remission (3 out of 9 patients). Neutrophils with reduced FcR111 expression showed more defective chemiluminescence and phagocytosis than neutrophils with normal FcRIII expression. Circulating myeloid cells from three patients in chronic phase, showing a normal percentage of CD16-positive neutrophils, were isolated and fractionated by discontinuous Percoll gradients. This study showed that CD16 appears at the stage of metamyelocyte, that band cells and segmented neutrophils display an identical pattern of membrane FcRIII, and that the fluorescence intensity shown by metamyelocytes is different from that displayed by more mature cells. The association between low FcRIII expression and function abnormality could be suggestive of a defect in CML neutrophil maturation.

Polymorphonuclear (PMN) motility is currently determined by a method using chemotactic chambers with micropore filters. PMN locomotive capacity is measured by evaluating the maximum distance travelled by the cells, or by counting the number of cells which have moved through the filter. Although in recent years some attempts to improve the analysis of chemotaxis data by computer-assisted systems have been made no technique has been shown capable of both measurement and correlation of these parameters in real time. In the present paper we describe an automated technique, based on a workstation capable of exploring microporous filters employed in chemotactic chambers and of measuring PMN motility in a reliable, reproducible, rapid manner, independent of the subjectivity of the operator. The measurement is carried out by custom software capable of undertaking computerized image analysis of microscopic fields acquired by a TV camera and of driving the motorized microscopic table. Depth of migration, cell distribution at each plane, correlation index of the random behaviour with the model described by a gaussian distribution and data about the patient or assay under study, are computed, displayed, stored in a data base and printed in real time.

The effect of lithium treatment on the impaired neutrophil chemotactic function of a patient affected by Shwachman-Diamond syndrome is reported. We found that (1) a cytoskeletal cellular defect seems to be involved in the impairment of neutrophil function (and perhaps of cellular secretion and chondrocyte function) in the syndrome; (2) intermittent neutropenia is always present in the syndrome, and (3) lithium seems capable, in addition to its capacity of inducing leukocytosis, of modulating leukocyte functions by modulating the microtubular system. The drug, at usual therapeutic dosage, was able to normalize neutrophil functions without side effects. As no therapy is available in this syndrome to date, our data suggest the therapeutic use of lithium in order to improve these cytoskeleton-mediated functions and the degree of neutropenia.

The effects of lithium on the microtubular and microfilament systems of neutrophil granulocytes is described. The phagocytic capacity of human neutrophils was reduced or inhibited by using vinblastine or cytochalasin B. Lithium at concentrations ranging from 0.5 to 2.0 mEq/l proved to be capable of antagonising the action of vinblastine, whereas the action of cytochalasin B was found to be unaffected by lithium. Based on the antagonising effect of lithium on vinblastine-induced inhibition of phagocytosis, it is suggested that lithium acts--at therapeutically achievable levels--on the microtubular system of neutrophils, possibly through modulation of cyclic AMP levels.

Serum levels of total cholesterol and triglycerides were studied in 202 patients affected by various hematological malignancies at the time of diagnosis. A hypocholesterolemia was found in 44% of patients affected by lymphoproliferative diseases and acute lymphoblastic leukemia, with an evident correlation with the clinical stage (5.7% of patients in nonadvanced stages, 67.8% in advanced stages). In acute and chronic myeloproliferative diseases, the overall incidence of hypocholesterolemia was 71%. In particular, a greater incidence of low cholesterol values was found in chronic myeloid leukemia and in idiopathic myelofibrosis than in polycythemia vera. No significant correlation was found in this group of diseases between the values of cholesterol and the main hematological parameters studied (WBC, number of circulating blasts, degree of splenomegaly, levels of hemoglobin, hematocrit). The incidence of significant alterations of triglycerides appeared negligible. It is thus possible to affirm that hypocholesterolemia constitutes an interesting biological aspect in hematological malignancies, and that total cholesterol could represent a parameter, even though secondary, in the follow-up of hematological neoplastic pathologies.

A patient with copper deficiency and renal failure was suffering from pancytopenia. Marrow examination showed cytoplasmic and nuclear vacuolizations of the erythroid and myeloid series. These abnormalities disappeared after oral copper therapy. Possible mechanisms causing the pancytopenia are discussed. Copper supplementation is needed for patients receiving dietary changes.

Leukocyte migration inhibition was studied by employing the agarose plate method. The inhibitory effect of blood sera from healthy subjects, from patients affected by lymphomas or other immunoproliferative diseases and from patients with diseases not involving the immuno-system has been investigated. No inhibitory effect was detected in the sera of patients affected by various pathology including four solid neoplasia. It was possible to demonstrate the presence of an inhibitory factor in the sera of patients affected by Hodgkin's disease as in sera of patients affected by other immunoproliferative diseases.

ADA (E.C.3.5.4.4.) and PNPase (E.C.2.4.2.1.) are enzymes which have been extensively studied by imnunologists ever since it was noted that a deficiency of either one of these enzymes could be associated with severe congenital immunodeficiency. Though the catalytic actions of these enzymes are sequential, the absence or the alteration of one of the two enzymes causes different types of congenital immunodeficiencies. ADA insufficiency causes a severe combined immunodeficiency while PNPase insufficiency alters delayed immunity (1,2). JVbreover it has been demonstrated that these enzymes are involved in lymphocytic ontogeny both in animals (3) and in man (4).

The effects of lithium carbonate on leukocyte functions in a case of chronic benign neutropenia are presented. Lithium was able to induce leukocytosis and to bring about increases in chemotaxis, marrow granulocyte reserve test and phagocytosis. After lithium interruption, leukocyte functions returned to initial values. Some hypotheses are advanced to account for lithium action.

The modifications in the concentration of circulating myoglobin have been studied by means of a radioimmunoassay in 15 cancer patients undergoing polychemotherapy including adriamycin. In 8 patients significant increases in myoglobin levels were found after injection of low doses of the drug (25-50 mg/m2). Moreover, a disturbance of the normal biorhythm of the protein was evident in 12 patients. Creatine kinase-MB was evaluated by means of a radioimmunoassay, but there was no relation between an increase in the isoenzyme and an increase in myoglobin. No ECG modifications were detected. These data indicate that the measurement of myoglobin may offer an indication of myocardial or skeletal muscle damage caused by adriamycin.

The presence of leukocyte inhibitory factor(s) (S-LIF) in the serum of patients with Hodgkin's disease (HD) has been investigated. It was possible to demonstrate the inhibitory activity in 50% of the patients by employing the agarose plate method. The PHA response in these patients has been found to be reduced. A linear correlation exists between this reduction and the presence of S-LIF in the sera. In order to assess some biological properties of these sera, chemotaxis and phagocytosis of normal cells in the presence of sera from patients with HD have been investigated and both these leukocyte functions have been found to be reduced. The inhibitory capacity was detectable in the same sera which exhibited S-LIF activity and a close linear correlation could be shown. In order to assess whether S-LIF had the same properties of in vitro produced LIF, two monosaccharides were added: l -fucose, capable of blocking the inhibitory activity of LIF, and glucose, which does not block this activity. Surprisingly the addition of l -fucose and glucose greatly enhanced the inhibitory activity of S-LIF. These observations clarify the relationship between the presence of S-LIF and the immunological status in patients with HD and give some indications of the characterization of this (these) factor(s).

Thymostimulin (TP1 — Serono), which has several immunobiological activities in vivo and in vitro, is a pool of thermostable polypeptides extracted from calf thymus and has been used in the treatment of primary and secondary immunodeficiencies (1, 2). However, the immunobiological properties and the mechanism of action of this cellular extract is still not completely understood. For this reason, we have studied the activity of thymostimulin on the intracellular levels of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) enzymes in peripheral blood lymphocytes of patients with hematological malignancies. We also hoped to reveal the effects of thymostimulin on lymphocytes from cord blood and from elderly patients. In fact, these enzymes have important roles in both lymphocyte ontogenesis (3, 4) and in the maintenance of immunological functions (5, 6).

Multiple sclerosis (MS) is postulated to be an immunopathologically mediated disease. This concept is supported by the finding of abnormally distributed peripheral blood T-cell subsets and a decreased T-suppressor function. Thirty-seven MS patients have been selected according to the criteria for definite MS. Fluorescein- or phycoerythrin-conjugated monoclonal antibodies have been used to define different lymphocyte subsets: CD4+, CD5+, CD8+, CD19+, CD38+, CD45RA+, CD4+CD45RA+, CD19+CD5+, CD8+CD38+. In relapsing-remitting (RR)-MS patients a significantly decreased percentage of CD19+ cells and in progressive MS patients a significantly increased percentage of CD19+CD5+ cells have been found. During a relapse in RR-MS, a significantly decreased percentage of CD4+CD45RA+ cells and a significantly increased percentage of CD8+CD38+ cells have been observed. Moreover, in RR-MS patients a significantly increased percentage of CD38+ cells and significantly high IgM amounts have been found. The increased percentage of CD19+CD5+ and CD38+ cells (together with high IgM levels) and the reduced percentage of CD4+CD45RA+ lymphocytes could be related to an activation of both cellular and humoral immune response in acute MS.