F. Ambrogi

Immune Cell Imbalance in Major Depressive and Panic Disorders

We investigated subsets of peripheral immunologic cells in 12 drug-free patients affected by major depression according to DSM-III-R criteria, and who had recent evidence of somatic diseases. They were compared with 10 drug-free depressives, with 10 patients with panic disorder, and with 12 healthy volunteers, all without somatic disease. The immune subsets were measured by flow cytometry. The results showed that both groups of depressives had the same abnormalities in immune cells compared with the healthy volunteers or the panic disorder patients; in particular they presented a lower number of CD3+, CD8+ and HLA-DR+. The patients with panic attacks did not differ from healthy controls, except for CD4+ cells which were significantly lowered, even in comparison with the depressive groups. These data, although preliminary and in a small sample, suggest that some immune parameters may be influenced by the presence of a major psychiatric disorder.

Immunological alterations in adult obsessive-compulsive disorder

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 Sydenhams 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.

Lymphocyte subsets in multiple sclerosis A study with two-colour fluorescence analysis

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.

Detection of Eosinophils in Whole Blood Samples by Flow Cytometry

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, Hodgkins 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 vivo Effectiveness of Lithium on Impaired Neutrophil Chemotaxis in Shwachman-Diamond Syndrome

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.

Effects of Different Low-Frequency Electromagnetic Fields on Lymphocyte Activation: At which Cellular Level?

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.

Motility of rhG‐CSF‐induced neutrophils in patients undergoing chemotherapy: evidence for inhibition detected by image analysis

The motility of circulating neutrophils from seven patients affected by intermediate and high‐grade non‐Hodgkins lymphoma was investigated before and after rhG‐CSF administration (5μg/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.

FcRIII (CD16) expression on neutrophils from chronic myeloid leukemia. A flow cytometric study

FcRIII (CD16) expression on 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 FcRIII 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.

Cell-mediated immunity in breast cancer patients.

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 P < 0.05 - P < 0.001) than in the control group. The mean values of NK cells and B lymphocytes, the former parameter being highly significant (P < 0.001), 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, P < 0.05) 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 (P < 0.01 and P < 0.001 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 (P < 0.05) 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.

Serum Cholesterol and Triglycerides in Hematological Malignancies

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.