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.     

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.     

Action of Levamisole on E-Rosette Forming Cells and Serum Adenosine Deaminase in Hodgkin's Disease

The interaction of levamisole and/or adenosine in vitro an E-rosette forming cells (RFC) of healthy subjects and on low RFC of patients affected with various diseases was investigated. Levamisole did not enhance normal RFC. The drug enhanced low RFC in some patients (responders) but not in others (non-responders). Adenosine inhibited normal and low RFC and this inhibition could be reversed by levamisole.

Further, levamisole 150 mg 2 days a week was given to 12 Hodgkin's disease (HD) responders for 2 weeks, then 150 mg 1 day weekly for 2 1/2 months After one week of treatment RFC increased and the enhancement could be maintained during the whole treatment period.

Lastly levamisole in vitro and in vivo diminished serum adenosine deaminase activity in normal subjects and HD patients.     

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.     

Monocyte Locomotion in Anergic Chronic Brucellosis Patients: The In Vivo Effect of Ascorbic Acid

In 14 patients suffering from relapsing chronic brucellosis who were anergic to brucella antigens, we have studied peripheral blood monocyte random migration and chemotaxis against non-specific and specific leukoattractants, as well as plasma and monocyte ascorbic acid levels.

We found that all parameters studied, were significantly beneath normal, when compared to normal subjects.

After the oral administration of ascorbic acid at a daily dose of 1gr for 15 conseguetive days, random and directed migration against a non-specific stimulus (casein)returned to normal. Directed migration against disease associated leukoattractants (brucella melitensis and brucella abortus) antigens improved significantly, without reaching normal values.

We concluded that ascorbic acid supplementation might partially restore peripheral, monocyte function and help the monocyte-macrophage system to mount an effective immune response against chronicity of brucella infection.     

Levamisole treatment of oligospermia

On the basis of elevated levels of seminal plasma immunoglobulins and a decreased production of LMI in the presence of PPD, 6 patients (out of the idiopathic normogonadotrophic oligospermic group) were selected for a therapeutic trial with levamisole. The following regime of treatment was applied: 3 days on levamisole (150 mg per day) followed by a 10 day interval without treatment, for a period of three months. The sperm count increased in 5 out of 6 patients and motility increased in 2. At the same time a reduction in seminal plasma IgG levels was observed in all treated patients. A significant increase of EA rosettes was observed after treatment.     

The in vitro effect of thymic humoral factor and levamisole on peripheral blood lymphocytes in systemic lupus erythematosus patients.

The in vitro effect of thymic humoral factor (THF) and levamisole on E rosette-forming cells in the peripheral blood of seventeen patients with systemic lupus erythematosus (SLE) was studied. Patients with active disease showed a low number of E rosette-forming cells. A significant rise in the number of E rosettes was obtained after incubation with both THF and levamisole. No such effect was observed on lymphocytes from patients with inactive disease and normal controls. In seven patients, three with active disease and four with well-controlled disease, short-term cultures were performed. The effect of THF on E rosettes was found to be the same before and after the short-term cultures. Possible mechanisms, by which THF (on the one hand) and levamisole (on the other) may increase the number of E rosettes in vitro, are discussed.     

Concentric Membranous Bodies in Hepatocytes from a Patient with Systemic Lupus Erythematosus

Concentric membranous bodies (CMB) are described in the cytoplasm of hepatocytes in a liver biopsy from a patient with systemic lupus erythematosus (SLE). Histologically, the biopsy showed binucleated hepatocytes and rosette formation with lymphocytic infiltrates in portal fields. Hepatocyte cytoplasm showed focal fatty metamorphosis, focal staining for RNA with azure B at pH 4.0 (which was perchloric acid labile), diffuse alloxan-Schiff staining for protein, and PAS-positive, diastase-sensitive glycogen.

By electron microscopy one to three CMB were present in 10-15% of hepatocytes. CMB appeared as concentric, “fingerprint” or parallel arrays of particlestudded, membranous profiles. Other membranous configurations included mazelike forms. The space of Disse was dilated. Bile ductular changes included basement membrane thickening and redoubling and luminal bulging of ductular epithelium devoid of microvilli.

CMB have been rarely reported in human hepatoma and have not heretofore been observed in nonneoplastic human liver. Their appearance in hepatocytes in a patient with SLE may reflect an increase in protein synthesis during regeneration.     

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.     

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.     

TOLERABILITY OF NIMESULIDE AND PARACETAMOL IN PATIENTS WITH NSAID-INDUCED URTICARIA/ANGIOEDEMA

Previous studies evaluated the tolerance of nimesulide and paracetamol in subjects with cutaneous, respiratory and anaphylactoid reactions induced by nonsteroidal anti-inflammatory drugs (NSAIDs).

In this study we investigated tolerability and reliability of nimesulide and paracetamol in a very large number of patients with an exclusive well-documented history of NSAID-induced urticaria/angioedema. Furthermore, we evaluated whether some factors have the potential to increase the risk of reaction to paracetamol and nimesulide.

A single-placebo-controlled oral challenge procedure with nimesulide or paracetamol was applied to 829 patients with a history of NSAID-induced urticaria/angioedema.

A total of 75/829 (9.4%) patients experienced reactions to nimesulide or paracetamol. Of the 715 patients tested with nimesulide 62 (8.6%) showed a positive test, while of 114 subjects submitted to the challenge with paracetamol, 13 (9.6%) did not tolerate this drug. Furthermore, 18.28% of patients with a history of chronic urticaria and 11.8% of subjects with an history of NSAID-induced urticaria/angioedema or angioedema alone (with or without chronic urticaria) resulted to be intolerant to alternative drugs.

Taken together, our results confirm the good tolerability of nimesulide and paracetamol in patients who experienced urticaria/angioedema caused by NSAIDs. However, the risk of reaction to these alternative study drugs is statistically increased by a history of chronic urticaria and, above all, by a history of NSAID-induced angioedema.     

In vitro reversal of cellular unresponsiveness induced by levamisole.

Mononuclear cells from twenty-one patients with depressed cellular reactivity were assessed for the ability to produce leucocyte inhibitory factor (LIF) and to transform after PHA stimulation, in the presence or absence of levamisole. Cells from nineteen patients failed to produce significant lymphokines when stimulated with PHA alone, but after a prior 1-hr levamisole pulse normal amounts of LIF were produced. Unstimulated mononuclear cell supernatants from six patients showed LIF-like activity, which could be abolished or decreased in five of the six when the cells were initially treated with levamisole. Mononuclear cells from seven of twelve patients which failed to incorporate [3H]thymidine after PHA activation, showed an increased response after a 1-hr levamisole pulse. Unstimulated mononuclear supernatants from six patients inhibited the lymphoproliferative response of normal cells to PHA. After treatment with levamisole, however, the suppressive effect of these supernatants was decreased or abolished. In vitro levamisole treatment, therefore, not only restores cellular responsiveness in anergic patients but also restricts the uncontrolled release of inhibitory factors.     

Immunohistochemical and Ultrastructural Analysis of Bronchopulmonary Neuroendocrine Neoplasms. I. Carcinoids

Twenty-five strictly defined bronchopulmonary carcinoids were studied by light microscopic immunohistochemistry by the peroxidase technique for NSE (neuronspecific enolase), serotonin, and a broad spectrum of neuropeptides. Eighteen cases were also studied by electron microscopy.

Twenty-three of the twenty-five cases showed immunostaining for NSE; 24 cases displayed immuno-staining for at least two of the hormones tested for; the single case that failed to show hormonal immuno-reactivity was however positive for NSE and had granules by electron microscopy. Serotonin was the most frequently demonstrated hormone followed by bombesin, vasoactive intestinal peptide, gastrin, leuenkephalin, alphamelanocyte stimulating hormone, somatostatin, substance P, and calcitonin. In several cases, adjacent-step sections stained for different hormones strongly indicated immunoreactivity for more than one hormone in single neoplastic cells.

By electron microscopy, all 18 cases studied showed generally abundant neurosecretory granules, which, however, displayed considerable heterogeneity in their size, shape, and density. Twelve of these eighteen cases displayed evidence of squamous differentiation and 10 showed characteristic exocrine lumina.

The capability of single neuroendocrine tumors and single neuroendocrine tumor cells to produce more than one immunoreactive hormone is hereby amply confirmed; these broad capabilities are certainly reflected in the heterogeneous granule populations seen by electron microscopy. The synchronous presence of squamous and exocrine features in bronchopulmonary carcinoids indicates that they too are capable of multidirectional differentiation, which should not detract from their being regarded basically as well-differentiated neuroendocrine neoplasms. The clinical significance of strictly defining bronchial carcinoids is underscored by the fact that of 25 cases followed for 2-13 years, only one developed metastases 9 years postoperatively.     

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.     

Book Review

GENERAL IMMUNOLOGY by E. L. Cooper

ANTIBODY AS A TOOL The Applications of Immunochemistry, J. J. Marchalonis, G. W. Warr, Eds., A. Johnstone

Both of the above-mentioned works provide a vast array of immunochemical techniques, for the non-immunologist or the beginning immunologist (on a graduate level).

CLINICAL IMMUNOLOGY OF THE KIDNEY, J.B. Zabriskie H., Fillit H., Villareal Jr. E.L. Becker, Eds.

IMMUNE MECHANISMS IN RENAL DISEASES, N.B. Cummings, A.F., Michael

SERUM PROTEIN ABNORMALITIES (Diagnostic and Clinical Aspects), S.E. Ritzmann, J.C. Daniels, Eds.

IMMUNITY AND CONCOMMITANT IMMUNITY IN INFECTIOUS DISEASES P. Kallos, ed.

SEMINAR ON AUTIGEN-AMTIBODY REACTIONS REVISITED, C.A. Bell, Ed.     

Surface infrastructure of the Small Intestine Mucosa in Healthy Children and Adults: A Scanning Electron Microscopic Study with Some Methodological Aspects

Biopsy specimens of light microscopically (LM) normal small intestine mucosa from eight healthy, constitutionally short-statured children without signs of gastrointestinal disease and six healthy adults were studied by scanning electron microscopy (SEM) supplemented by transmission electron microscopy (TEM). The effects on surface morphology of various preparative procedures were also investigated, using small intestine mucosa from cats and rats.

Fixation with OsO₄-either alone, or following glutaraldehyde fixation-markedly changed the surface ultrastructure compared to that after glutaraldehyde fixation only.

By low power SEM, some differences were observed in the appearance of the small gut mucosa between adults and young children. In adults and in children above 3 years of age, the villi were usually shaped like fingers or leaves, but in infants, ridge-shaped villi predominated. The villi showed, however, a smooth surface in both infants and adults, and medium and high power SEM displayed similar pictures, irrespective of age; here the typical structural features of the normal small gut mucosa in humans were (1) distinct extrusion zones at the crests of the villi and almost no signs of enterocyte extrusion along the sides of the villi, and (2) regular enterocytes with polygonal, flat, apical surfaces covered by a thick gly-cocalyx that obscured the underlying microvilli.     

In vivo effects of recombinant-interferon-beta1b treatment on polymorphonuclear cell and monocyte functions and on T-cell-mediated antibacterial activity in patients with relapsing-remitting multiple sclerosis

Treatment with Interferon (IFN)-β has been proposed as a therapeutic approach in multiple sclerosis (MS) patients, mostly in view of its immunomodulating actions. At the same time, evidence has been provided that MS patients exhibit polymorphonuclear cell (PMN) deficits, which can explain the increased susceptibility to infections in these subjects. Here, in 28 patients with relapsingremitting (RR) MS under treatment with recombinant (r)-IFN-β PMN polarization and PMN and monocyte (MO) phagocytosis and killing, as well as T-cell mediated antibacterial activity, were evaluated before treatment and over a period of nine months of treatment.

Our results point out an enhanced rate of polarization (both “spontaneous” or N-formyl-methionyl-leucyl-phenylalanine-induced) in MS patients. After r-IFN-β_1b treatment the polarization rate was further increased. On the contrary, PMN and MO phagocytosis and killing were depressed in comparison to controls and values were further reduced by r-IFN-β_1b treatment.

In patients T-cell mediated antibacterial activity was decreased at TO and dramatically dropped in the course of r-IFN-β_1b therapy.     

Combined antiviral therapy reduces HIV-1 plasma load and improves CD4 counts but does not interfere with ongoing lymphocyte apoptosis

The progression of HIV-1 disease appears associated with an unregulated Fas-mediated apoptosis of lymphocytes that involves the activation of ICE protease and ceramide generation and antiviral therapy may not be fully effective in the absence of a relevant impact on apoptosis.

Six drug-naive HIV-1-infected symptomless patients with advanced immunodeficiency were treated with combined AZT and dd1 for 4 months; plasma HIV-1 RNA levels, the counts of CD4 cells, CD4 and CD8 apoptotic lymphocytes, Fas-positive cells and ICE-positive cells, and intracellular ceramide levels were measured at base-line and after 7, 45 and 120 days of treatment.

There was a prompt reduction in plasma viremia and a secondary increase in CD4 counts, but the treatment had no impact on apoptotic CD4 and CD8 lymphocytes, Fas-positive cells and ICE-positive cells, and on the intracellular levels of ceramide.

A discrepancy exists between the positive impact of combined AZT and ddl treatment on plasma viral load and CD4 counts and the lack of any effect on the process of lymphocyte apoptosis. We suggest to use the measurement of apoptotic lymphocytes as a surrogate marker to predict, in combination with viral load and CD4 counts, a large proportion of the clinical effect of antiviral therapy.     

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.     

Recurring Congenital Lesion of the Cheek

A 3-cm purple area was present on the left cheek of a newborn white female, and a biopsy showed atypical cells in fibroadipose tissue. The lesion increased in size and the infant was referred to the M. D. Anderson Hospital. By 6 weeks, the lesion formed an irregular 6-cm paranasal mass, compressing the nostril and depressing the corner of the mouth. It was soft and mobile with a nodular, erythematous, excoriated surface. Three small satellite masses were also present. Another biopsy was obtained, establishing the presence of a malignant neoplasm. Vincristine, adriamycin, and cyclophosphamide produced a rapid decrease in size of the lesion, and by 6 months it was considerably smaller and firmer. Total surgical excision was then performed. The well-circumscribed 3-cm nodule had a glistening, tan-gray cut surface. The wound was closed with a skin graft, and

By 11 months, the lesion had recurred as a firm, round mobile nodule in the premaxillary area, lateral to the previous surgical site. The reexcision specimen revealed a firm, pale tan, 2-cm mass with a lobulated cut surface. In view of the previous reqoonse, chemotherapy with vincristine, adriam ycin, and cyclophosphamide was reinstated, with modification of the dose and schedule. Clinical evidence of adriam ycin cardiom yopath y developed, the chil's condition deteriorated, and she died at the age of 18 months. Autopsy was not performed.