Reevaluation of the Effect of Levamisole in Chronic Brucellosis: In Vitro and in Vivo Effect on Monocyte Phagocytosis

The in vitro effect of levamisole on peripheral blood monocyte(P.B.M.) phagocytosis was studied in 32 patients with chronic brucellosis and 20 normal subjects. It was shown that levamisole enhances P.B.M. phagocytic capacity, not reaching however normal levels. A subgroup of 11 patients were treated with levamisole for 6 months and the drug effect on cellular and humoral immunity and monocyte phagocytosis was also studied. By the end of the 6 month treatment-study period, the following results were obtained: 1. six patients were symptom free while five had significantly improved. 2.T lymphocyte number and monocyte phagocytosis reached normal values. 3. Significant specific cellular immunity against both brucella antigens was noted. 4.B lymphocytes showed no significant changes. 5. Antiglobulin titers varied. These findings suggest that the good therapeutical effect of levamisole in patients with chronic brucellosis could probably be attributed to the enhancement of both T-cell function and monocyte phagocytosis.     

Immunotherapy in Chronic Brucellosis. Effect of Levamisole and Interferon; Mechanisms of Action and Clinical Value

Thirty two anergic patients with chronic brucellosis treated with a)interferon-alpha2b(group 1), b)levamisole (group 2) and c)conventional therapy(group 3) were studied. the effect of treatment on T lymphocyte blast formation in the presence of PHA, specific cell mediated immunity against brucella antigens, titers of brucella antibodies and clinical symptoms were evaluated T lymphocyte blast formation was shown to range in normal levels in all patients before treatment compared to 10 normal controls suggesting against a generalized impairment of cell mediated immunity. Titers of brucella antibodies were significantly decreased in group 1, almost significantly in group 2 and were significantly increased in group 3 at the end of treatment. A significant improvement of symptoms as well as production of leukocyte migration inhibition against brucella antigens were noted in both groups 1 and 2, in contrast to group 3. This response to treatment was however greater in group 1. These findings demonstrate that immunotherapy resulted in both clinical and immunological improvement and that interferon seems to be a more promising therapeutic approach of chronic brucellosis.     

Reevaluation of the Effect of Levamisole in Chronic Brucellosis: In Vitro and in Vivo Effect on Monocyte Phagocytosis

Abstract

The in vitro effect of levamisole on peripheral blood monocyte(P.B.M.) phagocytosis was studied in 32 patients with chronic brucellosis and 20 normal subjects. It was shown that levamisole enhances P.B.M. phagocytic capacity, not reaching however normal levels. A subgroup of 11 patients were treated with levamisole for 6 months and the drug effect on cellular and humoral immunity and monocyte phagocytosis was also studied. By the end of the 6 month treatment-study period, the following results were obtained: 1. six patients were symptom free while five had significantly improved. 2.T lymphocyte number and monocyte phagocytosis reached normal values. 3. Significant specific cellular immunity against both brucella antigens was noted. 4.B lymphocytes showed no significant changes. 5. Antiglobulin titers varied. These findings suggest that the good therapeutical effect of levamisole in patients with chronic brucellosis could probably be attributed to the enhancement of both T-cell function and monocyte phagocytosis.     

Immunological Consequences of Zidovudine Treatment in Control and Morphine or Methadone Treated Mice

The effects of acute and chronic zidovudine (AZT) administration on immunologic test responses of mice were studied. The effects of AZT administration combined with morphine or methadone treatment, were also studied separately comparing the effects of each drug.

We noted that AZT-treatment did not modify the T-lymphocyte subsets (L₃T₄/LyT₂ rate), whereas morphine-treatment and AZT plus morphine treatment decreased the percentage of T helper cells.

Acute and chronic AZT-treatment increased Natural Killer cell (NK) activity and also recovered the decreased NK cell activity produced by morphine-treatment.

AZT-treatment, morphine-treatment, AZT plus morphine treatment and AZT plus methadone treatment strongly depressed the phagocytic physiological activity of Polymorphonuclear leukocytes (PMNs).

Another evidence of immunologic responsiveness against AZT was the reduction of the mitogenic and antigenic response of lymphocytes.

These results suggest a negative role of AZT-treatment especially on phagocytic activity and confirms a depressive effect of morphine-treatment on several immune functions studied. Furthermore, there is no indication of additive or synergistic toxic effects of AZT, morphine and methadone on the immune functions above that seen with each of these drugs when tested alone.     

Impairment of Non-Specific Immunity in Patients in Persistent Vegetative State

In fourtheen patients in persistent vegetative state (PVS) immune responsivenes was investigated. In particular, we studied the relationship between brain lesions following traumatic injury and immune system. In this respect, phagocytosis and killing of Candida albicans by polymorphonuclear cells (PMN) and monocytes were tested. In addition serum levels of Interferon-γ (IFN-γ) were evaluated.

The patients come out fiom PVS by 3-4 month were used as control group.

Data shown a profound impairement of phagocytosis and killing of monocytes and low serum levels of IFNγ when compared with normal values.

Taken together, these findings suggest that brain lesions, may affect non-specific immune response.     

In Vitro Effect of Interferon Alpha-2b on T Lymphocyte Transformation and Leukocyte Migration Inhibition in Patients with Chronic Brucellosis

The in vitro effect of IFN-a on lymphocyte transformation and specific immune response against Brucella antigens was studied in 33 patients with chronic brucellosis and 10 normal controls. The following immunologic in Aditro tests were applied: PHA activated lymphocyte transformation test using Bromodeo-xyuridine and a monoclonal antibody in the presence and abscence of 50 and 100 IU IFN Alpha-2b and leukocyte migration inhibition test against Brucella antigens in the presence and abscence of 100 and 500 IU of IFN Alpha-2b. Patients were further divided to 2 subgroups according to a positive or negative migration inhibition test.

Our results showed that T lymphocyte transformation was similar in patients and controls and that the addition of 50 IU IFN resulted in a significant increase of transforming cells whereas in the concentration of 100 IU IFN only anergic patients and controls responded positively. IFN also resulted in a significant leukocyte migration inhibition only in anergic patients and controls. These findings suggest that the chronic infection is not due to a generalized cellular immunodeficiency state and that IFN Alpha-2b might be a promising therapeutic approach in anergic patients.     

CPH-86, A Highly Purified Podophyllotoxin, Efficiently Suppresses in Vivo and in Vitro Immune Responses

Podophyllotoxin, a component of the plant resin Podophyllin, has been used as a clinical drug for many years. Recently it has been highly purified under the denomination of CPH-86.

We here demonstrate that extremely low doses of the compound efficiently inhibit antibody responses to SRBC and prolong allogeneic skin graft survival in mice.

In vitro immune reactions, such as mitogen and alloantigen induced proliferation and development of cytotoxic T cells, are also suppressed in a dose dependent manner.

This effect does not seem to be due to direct cellular toxicity or to a shift in the kinetic pattern of the responses.     

Adhesion Molecules in Gonarthrosis and Knee Prosthesis Aseptic Loosening Follow-up: Possible Therapeutic Implications

The involvement of the synovium is common in phlogistic processes of various joint diseases. A part from synoviocytes and the other cells in the synovial tissue, circulating cells recruited from peripheral blood also participate in the phlogistic process. The increased expression of adhesion molecules on both circulating and endothelial cell surface may further this recruitment.

We studied 15 patients affected by serious gonarthrosis requiring a prosthetic implant (GPI) and 7 with knee prosthesis aseptic loosening (KPL) to evaluate adhesion molecule expression and phlogistic infiltration in the synovium using immunohistochemistry and microscopic analysis. As control we studied 10 subjects affected by degenerative meniscopathies undergoing a selective arthroscopic surgical meniscectomy.

Analysis with Kruskal-Wallis test showed no statistical significant differences in the expression of CD54, CD11a, CD11b and CD18 in three groups examined. The model of variance analysis (Friedman test), showed that CD54 expression is greater in patients with GPI and KPL in comparison with the other molecules.

Adhesion molecules and their functions are important in arthropathies not only because their evaluation can allow us to identify the degree of inflammation and to predict its evolution, but also because pharmacological control of their expression could have important therapeutic implications.     

Influence of Lead Acetate on Hypersensitivity Experimental Study

Recent studies showed that lead acetate has an important immunotoxicity for the phagocytic activity as well as humoral and cell-mediated immunity.

We studied the influence of lead acetate on immediate and delayed hypersensitivity. The lead acetate exerts an important action on hypersensitivity reactions whether on rat mast cells de-granulation (immediate hypersensitivity) or on contact hypersensitivity.     

Study of CTL and Lak Contacts to Target Cells After Treatment with Mitomycin C and Adriamycin.

We tried to understand the role of Mitomycin C and Adriamycin in the increased killing of target cells by Cytotoxic T lymphocytes (CTL) and lymphokine activated killers (LAK).

For this purpose, we used an objective method allowing quantitative evaluation of the roughness of cell contours on electron micrographs. We compared the folding of the membranes of LAK and CTL as well as conjugated targets exposed to different treatments.

We demonstrated first that CTL and LAK displayed similar morphological patterns : the killer cells were more villous than targets in the free areas, and second that the former cells exhibited significant smoothing on the areas of contact with these targets.

These results suggest that the binding process (as distinct from the recognition step) is dependent on killer properties which are the same in CTL, LAK and probably NK cells.     

Effects of Ethinylestradiol on the Course of Spontaneous Autoimmune Disease in NZB/W and Nod Mice

Sex hormones affect (auto)immune responses in various ways. Investigations of the effects of estrogens have produced contradictory results.

We studied the effects of gender, gonadectomy and of (supra)physiological doses of (the orally active) ethinylestradiol (EE) in two spontaneous autoimmune disease models: the NZB/NZW F1 and NOD mice. In both models we confirmed the female preponderance and the aggrevating effects of gonadectomy in males but not in females.

The accelerated mortality found in NZB/W mice treated with supraphysiological doses of EE was not associated with increased proteinuria, increased IgG-type anti-DNA levels or increased mononuclear cell infiltrations in the submandibular gland. In contrast, we found a severe reduction in body weight and in the weights of various organs (indications of toxicity), and a decrease rather than an increase in proteinuria and in mononuclear cell infiltrations (indications for autoimmunity). Physiological doses of EE did not significantly affect disease symptoms.

In the NOD model a near-physiological, non-toxic dose of EE did not cause consistent changes on immunological disease symptoms either.

Therefore, we conclude that the sexual dichotomy in spontaneous autoimmune models is due to protective effects of androgens and that the mortality by estrogens is due to toxic effects rather than accelerated autoimmunity.     

Axonal enlargements (meganeurites) in neuronal ceroid lipofuscinosis (NCL)

Meganeurites have been described by light and electron microscopy in several neuronal lipid storage diseases. In neuronal ceroid lipofuscinosis (NCL), they appear to be less frequent.

We present the first combined Golgi-electron microscopy study of a patient with NCL, which shows that the axons from neurons with meganeurites arise from their distal poles.

Our results reinforce the assumption that neuronal malfunction in this disorder, as in other neuronal storage diseases, is also due to the presence of such meganeurites in the area of the axon hillock-initial axon segment.     

The Effect of Levamisole on Peripheral T-Lymphocytes of Patients with Malignant Lymphomas

The effect of various concentrations (0.015-10 μg/ml) of Levamisole (LMS) on the peripheral lymphocytes of patients with malignant lymphomas (ML) and normal donors was investigated in vitro. The parameters studied include : E rosettes forming cells (total T lymphocytes), active E rosettes (early T lymphocytes) and DNA synthesis induced by pitogens PHA and Con A.

LMS improved significantly lymphocyte response both in patients with ML and normal donors when the cells were stimulated by Con A. In both groups no significant effect was observed on the response to PHA nor on the percentage of E-rosettes, whereas the mean number of active E rosettes was significantly increased on all concentrations of the drug.

While in the normal subjects a positive statistical correlation between active E-rosettes and Con A response was observed, in patients with ML an inverse correlation was found. This latter correlation was partially reversed by LMS.     

Selective Platelet Immune Retention

Inbred rats were immunized with Freund's Adjuvant containing proteins of Mycobacterium tuberculosis, Mycobacterium butyrium, or dinitrochlorobenzene (DNCB). Native arterial blood was then passed over glass beads coated with those antigens. The platelet retention on the glass beads was measured.

Rats immunized with Complete Freund's Adjuvant developed accelerated platelet retention on beads coated with protein derivatives of tuberculosis (EkD) after just 18 seconds of blood flow. Rats Immunized twice maintained selective retention longer than those immnunized once. The test animals developed no cutaneous hypersensitivity nor precipitin lines on Ouchter-long gels against PPD. Rats sensitized to DNCB had accelerated platelet retention on MCB-coated beads. Results were temperature and complement dependent, and antigen-specific. Heparin caused a dose-dependent inhibition of the accelerated retention.

PYD potentiated the UP-induced aggregation of platelet-rich plasma (YW) from immunized rats.

These experiments suggest that platelets react selectively to antigens in the intravascular spaces in immune reactions.     

Secretory Leukemic B Cells Express T Cell Markers In Vitro A Phenomenon Suppressed by TPA

Immunological and biochemical markers of leukemia/lymphoma cells have provided valuable insight into hematopoietic malignancy and normal differentiation. The general assumption is that as early lymphoid cells become conmitted towards terminal differentiation they lose their capacity for bimodal differentiation and cells become restricted to B or T cell development and function.

We have observed that phenotypically “late” leukemic B cells close to secretory stage can spontaneously expressmature T cell antigens (T11, T4 and T8) after culture in vitro. In further studies of these cells, it was found that the biochemical marker Lactate Dehydrogenase (LD) follows the intermediate pattern expressed by thymocytes rather than that of typical B cells. The expression of T cell antigens can be blocked by incubating these cells with the phorbol ester TPA (12-0-tetradecanoyl phorbol 13 acetate) which promotes unidirectional B cell maturation to plasmacytoid cells in a way that mimics normal B cell differentiation. These observations indicate that presecretory malignant B cells are still programmed to express T cell biochemical and antigenic markers and this expression can be influenced by environmental conditions in vitro.     

Immunohistochemical and Ultrastructural Analysis of Bronchopulmonary Neuroendocrine Neoplasms. II. Well-Differentiated Neuroendocrine Carcinomas

We have attempted to characterize a group of bronchopulmonary neoplasms that share certain structural features with true carcinoids but appear distinctly more pleomorphic and behave far more aggressively. In reviewing our files from 1973 to 1982, 11 such neoplasms were identified; the original diagnoses were “atypical bronchial carcinoid” (3 cases), “malignant carcinoid” (1 case), “bronchial carcinoid” (3 cases), “peripheral carcinoid” (2 cases), and “peripheral oat cell carcinoma” (2 cases).

Of the 11 neoplasms, 5 were central and 6 were peripherally located. At presentation, 7 patients had lymph node rnetastases and 1 had a distant metastasis. No patient had a conventionally defined hormonal syndrome; however, 2 patients had a history of episodic flushing, one of which was associated with diarrhea.

All cases were studied by light microscopy and light microscopic immunohistochemistry for NSE (neuron-specific enolase), serotonin, and broad-spectrum neuropeptides. Five cases were studied by electron microscopy. By light microscopy, the tumors were composed of solid clusters of polygonal to fusiform cells in an evident organoid arrangement. Foci of glandular and/or squamous differentiation were seen in 7 cases. Pleomorphism was moderate and mitoses were readily found. Focal necrosis was seen. By immunohistochemistry, 10 cases expressed NSE immu-noreactivity. All cases demonstrated hormonal immu-noreactivity; in 9 cases, immunoreactivity for more than one hormone was observed. The hormones most frequently expressed were serotonin, bombesin, gastrin, leu-enkephalin, and ACTH.

By electron microscopy, all cases studied contained heterogeneous populations of neurosecretory granules; the latter, however, were not abundant and tended to aggregate either in the basal pole of the cells or, more frequently, in interlacing “dendritelike” cytoplasmic processes. Aggregates of intermediate filaments were frequently seen. Basal lamina deposition was seen but gaps and larger areas of discontinuity were frequent.

We believe that these neoplasms constitute a distinct pathologic entity for which the term “well-differentiated neuroendocrine carcinomay” has been proposed. Clinically, these tumors merit special attention since they are demonstrably more aggressive than true carcinoids but are distinctly less malignant than the intermediate or small cell variants of neuroendocrine carcinoma.     

Tannic Acid Binding of Cell Surfaces in Normal, Premalignant, and Malignant Squamous Epithelium of the Human Uterine Cervix

Alterations in tannic acid (TA) binding capacity of cell surface carbohydrates in normal, premalignant, and malignant squamous epithelium of the human uterine cervix have been studied using electron microscopic visualization in combination with microdensitometric evaluation.

While in normal epithelium there is distinct binding in four to five cell layers of the deep intermediate zone, cells of carcinoma in situ and invasive cancer lesions lack TA binding. In moderate dysplasia an intermediate reacting pattern is found.

Deep intermediate cells in areas bordering the carcinoma in situ lesions do not show any binding, although their ultrastructure cannot be distinguished from similar cells in normal tissue.

The TA deposition within the deep intermediate zone is probably related to the presence here of glycoprotein-containing membrane-coating granules.

The finding that TA binding discriminates between cells in normal squamous epithelium and morphologically normal cells in juxtaposition with lesional areas in premalignant and malignant epithelium opens the possibility for a more reliable cytologic diagnosis of cervical epithelial neoplasia.     

Some immunological aspects of patients with rhinitis in Lebanon

Background: Hitherto immunological determinates in Lebanese patients with rhinitis have not been investigated.

Objective: To identify causative allergens in Lebanese patients with allergic rhinitis and determine possible correlation's among serum allergen specific antibody, polyclonal IgE, IL-4, IL-5 and peripheral eosinophil levels.

Methods: One hundred and thirteen patients with a long lasting history of nasal obstruction, rhinorrhea, sneezing and nasal itching were investigated. Serum allergen specific antibodies using a panel of 10 potential allergens, IL-4 and IL-5 levels were determined by enzyme immunoassays. Polyclonal IgE levels were estimated by an immunochromatographic assay and eosinophil counts by a Coulter STKS counter.

Results: Based on the presence of serum allergen-specific IgE antibodies, 74 patients were considered to have an allergic etiology. Polyclonal IgE levels were elevated in 41 of the 74 allergic rhinitis patients while the other 33 patients had normal serum levels. In the remaining 39 specific IgE antibody-negative patients, 32 had normal, and 7 had elevated, polyclonal IgE levels. IgE specific antibodies to more than one allergen were detected in 59 of 74 patients. The most common causative allergens were mite, Dermatophagoides pteronyssinus, Dpt (83.8%) and Dermatophagoides farinae, Df (78.4%). Analysis of the data indicated that elevated polyclonal IgE levels correlated with the concentration of serum specific IgE antibodies and the number of the detected causative allergens per patient. Fifty-nine of 74 allergic rhinitis patients had elevated IL-4 levels and 44 had elevated IL-5 levels. The number of allergic patients with both elevated IL-4 and IL-5 levels was 24. Finally, only 9 allergic rhinitis patients had peripheral eosinophilia.

Conclusion: Mite Dpt and Df were the most common causative agents of allergic rhinitis in the Lebanese group studied. A prerequisite for Specific Immunotherapy is the identification of the causative allergen. Determinations of polyclonal IgE level and peripheral eosinophil count alone, as an aid to diagnosis are insufficient and may be misleading. On the other hand, determination of all the parameters studied in conjunction appears to be of diagnostic value.     

Liposomes in Immunology: Multilamellar Phosphatidylcholine Liposomes as a Simple, Biodegradable and Harmless Adjuvant Without any Immunogenic Activity of its own

Multilamellar phosphatidylcholine liposomes were coated with the antigens human serum albumin (HSA) or bovine gamma globulin (BGG) simply by suspending the liposomes in a solution of the antigens.

Antigen coated phosphatidylcholine liposomes showed nearly the same adjuvant activity after intravenous injection as liposomes of a more complex phospholipid composition.

Since phosphatidylcholine liposomes are biodegradable, harmless, easily obtainable, have no immunogenic activity of their own and may be administered intravenously, they seem to be a promising immunoadjuvant.     

The Secondary Immune Response Against Liposome Associated Antigens

In the present experiments, the secondary immune response against antigens is studied after priming with liposome associated antigens and booster injections with the antigen alone, in order to study the effect of liposomes on the generation of immunological memory against the associated antigens.

Liposomes show adjuvant activity with respect to both the primary and secondary immune response against associated human serum albumin (HSA). When the injected dose of liposome associated HSA was too low to elicit a primary immune response, generation of immunological memory against the antigen could not be detected. Horse radish peroxidase (HRP) associated with liposomes did not elicit a primary immune response, but immunological memory against the antigen was established.