In vitro study of the binding of antiribonucleoprotein antibodies to the nucleus of isolated living keratinocytes.

The general accepted dogma is that antibodies do not penetrate living cells. However, this has recently been challenged for anti-ribonucleoprotein antibodies (anti-RNPab). We have studied here the "penetration" of trypsin isolated keratinocytes in vitro, using indirect immuno-fluorescence and immuno-peroxidase techniques. 70% (+/- 22) of a living keratinocyte cell suspension showed nuclear speckled staining when incubated with anti-RNP sera but only 9.5% (+/- 4) were stained with an anti-DNA antiserum (homogeneous pattern). A suspension of dead keratinocytes gave similar percentages with both anti-sera (89.5 [+/- 8] and 76.6 [+/- 18] respectively). The penetration of anti-RNPab into the nuclei of living epidermal cells increased gradually during the first hour of incubation without a parallel increase in the deadh rate measured by trypan blue dye exclusion. There was still high percentage of stained cell even after high dilution (1/1000) of anti-RNP sera. However, the percentage markedly decreased after previous incubation of the cells with increasing concentrations of concanavalin A. No decrease was obtained with dead keratinocytes in the same conditions. After they had been incubated with anti-RNPab, the epidermal cells were still capable of adhering to culture flasks and of incorporating labeled precursors. Only 4% of the epidermal cells in suspension were able to form rosettes with antibody coated erythrocytes, and were thus bearing receptors for the Fc fragment of IgG. These results strongly suggest that anti-RNPab penetrated living epidermal cells, but not through Fc receptors as reported for mononuclear blood cells.

Modifications in tissue histamine levels in mast cell-deficient mice (W/Wv) and in their littermates (Wv/+, W/+ and +/+) grafted with a methylcholanthrene-induced fibrosarcoma: correlation with tumour rejection.

There is evidence that mast cells and their degranulation products are involved in resistance against tumours. Previously, we have shown that tumour incidence and growth were inversely correlated with basal histamine levels, i.e. mast cell numbers, in tissues of W/Wv (mast cell-deficient), Wv/+ (partially mast cell-depleted), and +/+ (mast cell-sufficient) mice, and that histamine levels were increased in numerous tissues of tumour-bearing animals, including C57BL/6 and C3H mice, Sprague-Dawley and Commentry rats. The aim of this work was to analyse the incidence and growth of a grafted tumour (fibrosarcoma MC-B6-1) in W/+ mice, as compared with W/Wv, Wv/+ and +/+ mice, and to study the modifications in tissue histamine levels in W/+, W/Wv, Wv/+ and +/+ tumour-grafted mice, in order to determine whether or not these modifications were correlated with resistance to tumours. We report confirmation that tumour incidence and growth are inversely correlated with basal tissue histamine levels in W/Wv, Wv/+, and +/+ fibrosarcoma-bearing mice. However, in W/+ mice (normal tissue histamine levels), tumour incidence was the same as in Wv/+ mice. Histamine levels in tissues of W/Wv, Wv/+, W/+ and +/+ tumour bearing mice were not significantly different from those in controls. They were higher in some tissues of Wv/+ mice rejecting the tumour than in Wv/+ mice not rejecting the tumour. However, in W/+ and +/+ mice, histamine levels were not significantly different, and even tended to be lower in most tissues of mice rejecting the tumour than in mice accepting the tumour. Overall, these results suggest that resistance to tumours cannot be ascribed solely to mast cells, and that other mechanisms may also be involved. Thus, further experiments are needed to clarify the exact role of mast cells and mast cell-derived mediators and cytokines in the defence against tumours.

Calcium and bicarbonate ions mediate the inhibition of mast cell histamine release by Avène spa water.

Avène spa water (ASW) inhibits the histamine release induced in mast cells by substance P; the inhibition is reversed by EDTA and by EGTA. Calcium and magnesium ions, the major cations present in ASW, were experimented in simple saline solutions in the presence of various counter-ions. Only calcium salts inhibited the peptidergic stimulation, with different potencies ruled by the nature of the counter-anion (HCO3- > Cl- > SO4(2)). On a Ca2+ concentration basis, ASW was, however, more inhibitory, suggesting that other compounds present in ASW potentiate the effect of calcium.

Dynamic redistribution of concanavalin A binding sites on isolated guinea pig keratinocytes.

Modifications in the distribution of the binding sites for concanavalin A (Con A) were studied on trypsin isolated living guinea pig keratinocytes. Fluorescein-labelled Con A was used and the in vitro procedure has included short-term cultures, experiments at 37 degrees C and 4 degrees C, the study of colchicine and vincaleucoblastine effects. It was possible to induce different patterns of staining corresponding to distinct redistribution of Con A binding sites; the distinct redistribution was correlated to the effects of Con A, colchicine and vincaleucoblastine. These findings demonstrated that the system used was appropriate to the study of some dynamic events on the keratinocytes membranes.

Cellular activation products in osteoarthritis synovial fluid.

In order to address the issue of the role of mast cells (MC) and nitric oxide (NO) in rheumatic synovial-fluid diseases, synovial fluid (SF) collected from the knee of patients with osteoarthritis (OA), articular chondrocalcinosis (ACC) or rheumatoid arthritis (RA) was examined for the levels of mast cells (MC), histamine, tryptase, phospholipase A2 and nitrite. MC counts were found to be elevated in the SF of OA patients as compared with RA patients. Histamine content in SF parallelled the number of MC. Tryptase levels were elevated in OA in comparison to RA and ACC, but the difference was not statistically significant. Identical PLA2 levels were recorded among the 3 groups. Nitrite concentrations were also higher in SF from OA patients as compared to RA patients. These results suggest that mast cells (MC), in association with various inflammatory cells, may contribute to inflammation and cartilage breakdown in osteoarthritis (OA).

Effect of a protein bound polysaccharide (PS-K) on tumor development and infections in splenectomized rats and mice.

Daily oral administration of PS-K, an immunomodulating agent, was found to improve survival rate in tumor bearing mice and rats and to decrease susceptibility to infection in mice. Surprisingly, splenectomized tumor bearing rats and mice responded better to PS-K treatment as measured by survival rate, tumor size and immunosuppressive properties of serum. The mechanism of PS-K antitumor activity in both experimental systems might be due to the alteration of splenic immune suppressive mechanisms during tumor growth in the host. On the other hand, host resistance to acute infection was found to be less dependent upon the presence or absence of spleen suggesting a different mechanism of PS-K activity in acute infectious disease.

Tissue histamine levels in male and female mast cell deficient mice (W/Wv) and in their littermates (Wv/+, W/+ and +/+).

Histamine levels have been determined in nine tissues of four kinds (W/Wv, Wv/+, W/+, +/+) of WBB6F1 male and female mice, and mast cells have been counted in the skin and the fundus of W/Wv and +/+ mice. By comparison with +/+ mice (1) the association of the two alleles Wv and W induced a drastic decrease in tissues histamine levels and in mast cell number. (2) The allele Wv alone induced a marked decrease in tissue histamine levels but less important than the two alleles W and Wv. (3) In contrast, the allele W alone induced at least in some tissues, an increase in tissue histamine levels. In W/+, Wv/+ and +/+ fertile mice, tissue histamine levels were higher in females than in males. In contrast, in W/Wv mice which are sterile, no difference between males and females was observed.

Alterations in rat epidermis provoked by chronic vitamin D deficiency.

Long-term effects of vitamin D deficiency on epidermis were studied using histometric techniques, [3H]thymidine incorporation into DNA (labeling index), estimation of epidermal acid phosphatase activity, and one-dimensional gel electrophoresis of keratin proteins. The decrease in epidermal thickness due to a reduced number of granular cell layers and a lower level of epidermal acid phosphatase activity were observed in vitamin D-deficient rats. The number of nuclei in the basal layer was increased. No changes in labeled index due to chronic vitamin D deficiency or to 'single injection of 1,25-dihydroxycholecalciferol to vitamin D-deficient rats were observed. A comparative study of the keratin composition revealed differences in the keratin polypeptide pattern: vitamin D-deficient epidermis specifically lacked two low-molecular-weight components and presented several quantitative differences among other keratin polypeptides. The changes in epidermal morphology and metabolism that took place with vitamin D deficiency were independent of plasma calcium levels because similar modifications were present in vitamin D-deficient but normocalcemic rats (fed a diet rich in calcium and supplemented with lactose). These findings suggest that vitamin D may be one of the important factors for maintaining normal epidermal structure and metabolism through an effect on cell differentiation and formation of granular cell layers. They offer the possibility of using epidermal modification as an additional marker of vitamin D deficiency.

Noninvolvement of histamine and prostaglandins in the dermatosis of magnesium-deficient hairless rats.

Hairless rats receiving a deprived Mg2+ diet developed long-lasting inflammatory dermatosis which was not influenced by treatments with mepyramine, cimetidine, aspirin or indomethacin. Dermatosis did not appear in deprived rats treated with intraperitoneal injections of Mg2+, or with dexamethasone. Histamine and prostaglandins did not seem to be involved in this dermatosis. The possible role of leucotrienes is discussed.

Clinical improvement in advanced cancer disease after treatment combining histamine and H2-antihistaminics (ranitidine or cimetidine).

In a randomized study 31 patients with advanced cancer disease in whom classical anticancer therapy had been abandoned received a daily combination of subcutaneous histamine and oral H2-antihistaminics. In 27 patients, treatment induced a marked clinical improvement as shown by a large rise in performance status (Karnofsky scale). Ten patients were still alive 3-14 months after initiation of treatment. Average survival in the 31 treated patients (172 +/- 113 days) was significantly longer than in 34 non-treated patients with similar advanced cancer (26 +/- 16 days, P less than 0.00001). In six treated patients, the size of liver and lung metastases decreased. Histamine was perfectly tolerated up to 4 mg/day.

Release of mast cell mediators and nitrites into knee joint fluid in osteoarthritis--comparison with articular chondrocalcinosis and rheumatoid arthritis.

In order to address the issue of the role of mast cells and nitric oxide (NO) in joint effusions occurring in the course of osteoarthritis (OA), synovial fluids collected from the knee of patients with OA, articular chondrocalcinosis and rheumatoid arthritis (RA) were studied for number of mast cells, and histamine, tryptase, phospholipase A2 and nitrite content. Mast cell counts are elevated in synovial fluid from OA patients when compared with RA. Histamine content in synovial fluid parallels the number of mast cells. Tryptase levels are elevated in OA in comparison with both other conditions, but do not reach the level of significance. Identical phospholipase A2 levels are recorded in three groups. Nitrite concentrations are also higher in synovial fluid from OA patients when compared with RA patients. These results suggest that mast cells in association with various inflammatory cells, may contribute to inflammation and cartilage breakdown in OA.

Nonspecific refractoriness to adenylyl cyclase stimulation in alveolar macrophages from infants with recurrent bronchiolitis.

Epidemiologic studies suggest an association between recurrent bronchiolitis in children younger than 3 years of age and diagnosis of asthma later in life. Bronchoalveolar lavages from 20 infants with recurrent wheezing and 18 nonwheezy control subjects were analyzed to determine whether alveolar macrophages of wheezy infants present abnormalities similar to those described in adults with asthma. Alveolar macrophages from both groups responded in vitro, in a concentration-dependent manner, to prostaglandin E2, salbutamol, and forskolin, drugs that increase cyclic adenosine monophosphate levels. However, alveolar macrophages from infants with recurrent wheezing accumulated less cyclic adenosine monophosphate than those from control subjects in response to all three stimulations. These results are in agreement with the reduced cyclic adenosine monophosphate response to different agonists demonstrated in leukocytes from patients with asthma, and suggest that this refractoriness could be one of the precipitating events in the development of asthma observed in a large proportion of infants who have had bronchiolitis.

Blood histamine levels in HIV-1-infected infants and children.

Blood histamine levels (BHL; ng histamine base/ml blood, mean +/- SEM) were determined in 118 infants born to HIV-1-infected mothers and in children infected after blood transfusion. In asymptomatic infected infants, BHL were not significantly different from HIV-1-negative infants born to HIV-infected mothers (54.1 +/- 5.6 vs. 56.4 +/- 3.5). BHL progressively decreased in the group with polyadenopathies (40.0 +/- 4.0), in the group with AIDS-related complex (28.8 +/- 2.9), and in patients with AIDS (20.4 +/- 0.9). The decrease in BHL was associated with a decrease in blood basophil number.

Comparison between number of basophils, blood histamine, and histamine release in cancer and noncancer patients.

In cancer patients with primary tumor with or without metastasis or metastasis alone, by comparison with healthy subjects and noncancer patients, the decrease in blood histamine levels is due to a decrease in total basophil number. These basophils have a normal content of histamine (1 to 2 pg per basophil) and are able to release histamine. The percentage of anti IgE-induced histamine release is not significantly different than in noncancer patients. The scarcity of basophils in cancer patients is not due to a leukopenia. Blood histamine levels and total basophil number are normal in patients after successful excision of their primary tumor without metastasis.

Blood histamine levels (BHL) in infants and children with respiratory and non-respiratory diseases.

BACKGROUND: Blood histamine levels are decreased after severe allergic reactions and in various chronic diseases. AIMS: To study blood histamine levels in infants and children with acute infectious and non-infectious, non-allergic, disease. METHODS: Blood histamine levels were investigated by a fluorometric method in infants and children admitted to hospital with bronchiolitis, non-wheezing bronchitis, acute infections of the urinary tract, skin and ear-nose-throat, gastroenteritis, or hyperthermia of unknown aetiology. Results of blood histamine levels and white blood cell counts were compared with those obtained for children recovering from benign non-infectious, non-allergic illnesses. RESULTS: As compared with control children, white blood cell numbers were significantly increased in children with acute infections of the urinary tract, skin and ear-nose-throat, and were significantly decreased in children with gastroenteritis. Blood histamine levels were significantly lower in children with gastroenteritis and hyperthermia than in children with other diseases and control children. It was not possible to correlate blood histamine levels and the number of blood basophils. CONCLUSIONS: BHL are significantly decreased in infants and children with acute gastroenteritis and hyperthermia of unknown aetiology. The mechanisms responsible for the decrease in blood histamine levels in children with gastroenteritis and hyperthermia are discussed.

Tissue histamine levels and mast cell numbers in tumour-bearing mice.

In C57BL/6 mice bearing the 3LL carcinoma and in C3H mice bearing the McC3 -1 fibrosarcoma (18th passage), the increase in skin histamine levels was correlated with the increase in mast cell number. The number of cells able to incorporate tritiated thymidine was proportional to the mast cell number. These results strengthen the notion that, in tumour-bearing rodents, the increase in tissue histamine is an active phenomenon.