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PubMed1990

Relationship of one form of human histamine-releasing factor to connective tissue activating peptide-III.

We have previously reported purification of three forms of histamine-releasing factors (HRFs) from mixtures of streptokinase-streptodornase stimulated human mononuclear cells and platelets with apparent molecular masses of 10-12, 15-17, and 40-41 kD (1989. J. Clin. Invest. 83:1204-1210). We have also prepared mouse MAbs against the 10-12-kD HRF (1989. J. Allergy Clin. Immunol. 83:281). Affinity-purified 10-12-kD HRF appears as a broad band upon polyacrylamide gel electrophoresis in the presence of SDS. We determined the NH2-terminal amino acid sequence of the top and bottom halves of this broad band. Sequence analysis revealed striking homology between this HRF and connective tissue activating peptide-III (CTAP-III), a platelet-derived 8-10-kD protein known to cause mitogenesis and extracellular matrix formation in fibroblast cultures. 19 of 21 NH2-terminal residues in the top half of the HRF band were identical to the NH2-terminal sequence of CTAP-III. 20 of 21 NH2-terminal residues in the bottom half were identical to the NH2-terminal sequence of neutrophil-activating peptide-2, which is derived from CTAP-III by proteolytic cleavage between residues 15 and 16. Purified CTAP-III also released histamine from basophils. Rabbit antiserum raised against either native or recombinant CTAP-III recognized affinity-purified HRF in immunodot blot assays, and MAb against HRF recognized CTAP-III in both dot blot and microtiter plate based immunoassays. These data demonstrate the first structural, functional, and immunologic relationship between one form of human HRF and a previously described cell product.

Amino Acid Sequence↗

PubMedDate not supplied

Evaluation of possible causes of delayed hypersensitivity impairment in cancer patients.

The separate roles of malnutrition, advanced age, and stage of tumor growth as causes of impairment of delayed hypersensitivity response (DHR) was studied in 111 patients with solid tumors and in 56 nonneoplastic control patients matched for age, anatomical site of disease, degree of illness, and nutritional status. Pretreatment DHR to recall antigens (tuberculin, Candida, streptokinase-streptodornase, trichophyton) and to dinitrochlorobenzene in cancer patients with 9% anergic, 43% hypoergic, and 48% normoergic; the distribution of DHR in controls was not significantly different. In cancer patients, the serum albumin level showed an inverse correlation with the stage of tumor (p less than 0.01) and a positive correlation with the DHR (p less than 0.001); the serum albumin level was also in the controls positively correlated with the DHR (p less than 0.01), indicating that malnutrition in neoplastic or benign disease may cause depression of DHR. In well-nourished controls, age was inversely correlated with DHR (p less than 0.05), showing that aging itself may be another relevant cause of depression of DHR. The results of this study indicate that DHR in patients with solid tumors is similar to the DHR of nonneoplastic patients if matched for age, sex, and nutritional status. DHR impairment in cancer patients appears to be caused mainly by aging and by malnutrition due to the advanced progression of cancer.

Adult↗

PubMed1978

The lymphocyte transformation response of fetal hemolymphatic tissue to mitogens and antigens.

Lymphocyte transformation responses to three different mitogens (phytohemagglutinin (PHA), pokeweed mitogen (PWM), and concanavalin A (Con A)) as well as four antigens (streptolysin O (SLO), keyhole limpet hemocyanin (KLH), streptokinase-streptodornase (SKSD) and tuberculin purified protein derivative (PPD)) were studied ontogenetically in 30 human fetuses ranging in gestational age from 6-19 weeks. In most organs lymphocyte responsiveness to Con A in human fetuses seemingly develops in concert with responsiveness to PHA and PWM. One 19 week fetus had an apparent antigen specific response to SLO with stimulation of cord blood and bone marrow lymphocytes. The same fetus also had mitogen responsiveness in bone marrow.

Antigens↗

PubMed2002

Delayed cutaneous hypersensitivity tests and lymphopenia as activity markers in sarcoidosis.

STUDY OBJECTIVES: To evaluate new and already known biological markers of activity in patients with sarcoidosis. DESIGN: A 10-year prospective clinical evaluation, including a battery of delayed cutaneous hypersensitivity tests (DCHTs) and other markers of activity. SETTING: Outpatient department of a university teaching hospital. PATIENTS: Forty patients with biopsy-proven sarcoidosis were prospectively evaluated every 6 months. In this study, only the visits that fulfilled the situation of active period (AcP) or of asymptomatic period (AsP) were taken into account. Twenty-one visits were considered to be in the AcP, and 26 were considered to be in the AsP. Seven patients were studied both in the AcP and the AsP. INTERVENTIONS: DCHTs and blood sample extraction every 6 months. MEASUREMENTS AND RESULTS: The mean diameter of the cutaneous wheal for each antigen (AG) was lower in the AcP group than in the AsP group (candidine, p < 0.0001; tuberculin, p < 0.0009; trichophytin, p < 0.02; streptokinase-streptodornase, p < 0.001). Also, the mean (+/- SD) diameter for the four AGs taken together was lower in the AcP group (2.3 +/- 4.2 mm) than in the AsP group (16.8 +/- 9.3 mm; p < 0.0001). The mean serum angiotensin-converting enzyme (S-ACE) value was higher in the AcP group than in the AsP group (p < 0.02). A low lymphocyte count and a percentage of the lymphocyte count (< 20%) also were detected more frequently in the AcP group than in the AsP group (p < 0.02 and p < 0.0001, respectively). CONCLUSIONS: DCHTs appear to be a simple, reliable, and easily performed marker of inflammatory activity in sarcoidosis patients. Furthermore, serum total and differential lymphocyte count and the S-ACE level proved to be useful inflammatory markers in this study.

Adult↗

PubMed2006

Tissue plasminogen activator combined with human recombinant deoxyribonuclease is effective therapy for empyema in a rabbit model.

OBJECTIVES: There have been no controlled studies to test the efficacy of tissue plasminogen activator (tPA) or recombinant human deoxyribonuclease (rhDNase) in the treatment of empyema. In vitro studies show that streptokinase without rhDNase does not liquefy empyemic material from rabbits. However, the combination of streptokinase and streptodornase and rhDNase have been shown to liquefy pus in vitro. The aim of this study was to determine if tPA or rhDNase, or a combination of the two, is more effective than saline solution in the treatment of empyema in rabbits. MATERIALS AND METHODS: Empyema was induced in rabbits using 10(9) Pasteurella multicoda organisms in infusion agar injected via a surgically placed chest tube. Once empyema was verified, a blinded investigator administered one of four treatments via the chest tube: 3 mL of saline solution; 4 mg of alteplase (recombinant tPA); 1 mg of rhDNase; or 4 mg of alteplase and 1 mg of rhDNase. The rabbits received a treatment every 12 h following the initial for a total of six treatments. The animals were killed at day 10, and the degree of empyema and pleural peel was scored macroscopically on a scale of 0 to 4. RESULTS: The combination group had a significantly lower mean empyema score (1.83 +/- 0.75) than did the saline solution group (3.86 +/- 0.38, p = 0.001), rhDNase group (3.17 +/- 0.75, p = 0.012), and alteplase group (3.71 +/- 0.49, p = 0.02) [+/- SD]. The total volume of pleural fluid was markedly higher in the alteplase and rhDNase combination group (142 +/- 79.1 mL) or the alteplase group (231 +/- 78.0 mL) compared to either the rhDNase group (0.8 +/- 1.6 mL) or the saline solution group (5.8 +/- 14.0 mL). CONCLUSION: The combination of alteplase and rhDNase is more effective in the treatment of rabbit empyema than either agent alone. The intrapleural injection of alteplase alone or in combination with rhDNase leads to the production of large amounts of pleural fluid.

Animals↗

PubMed1991

Immunoregulatory effects of phospholipase A2 (PLA2) on the proliferation of human lymphocytes.

Extracellular phospholipase A2 (PLA2) is a proinflammatory enzyme found especially in the inflammatory exudate to modulate blood flow to areas of antigen stimulation. In this study we found that PLA2 exerted a biphasic effect on the proliferation of phytohemagglutinin (PHA)-stimulated human mononuclear cells (PHA MNC). At low concentrations range from 0.001 to 1 U/ml, PLA2 enhanced the proliferation of PHA MNC (maximal increase was 37.0 +/- 5.67%). Conversely, at concentrations over 10 U/ml, PLA2 markedly suppressed the PHA-induced MNC proliferation (maximal decrease was 88.86 +/- 2.89%). PLA2 was non-toxic to lymphocytes after three days culture, unless the concentration was higher than 100 U/ml. The membrane polarization of PHA-stimulated lymphocytes was also increased by PLA2 at a low concentration. In addition, PLA2 displayed a similar effect on the proliferation of streptokinase-streptodornase (SK/SD) or allogeneic cell stimulated lymphocytes. The change of lymphocyte proliferation by PLA2, was parallel to the change of percentage of helper T cells. Furthermore--a CD4-rich population was proved more susceptible to PLA2 effect than a CD8-rich population. Para-bromophenacyl bromide (pBPB), an irreversible inhibitor of PLA2, abrogated the biphasic effect of PLA2 on PHA MNC proliferation. These results suggest that PLA2 plays a regulatory role on immune reactions by modulating the percentage of helper T cells.

Deoxyribonucleases↗

PubMed1987

Contribution of adherent cells and serum components to immune suppression in Kenyan visceral leishmaniasis.

Adherent cells and serum components from Kenyan patients with visceral leishmaniasis were examined with the view to evaluating their contribution to cell-mediated immune suppression. Mitogens (phytohemagglutinin and concanavalin A) and antigens (purified protein derivative, streptokinase-streptodornase, and leishmania) were used as stimulants. Compared to the controls, the contribution of serum components to suppression in presence of any of the mitogens and antigens was not significant. The same applied to adherent cells, except in the presence of leishmania antigen where adherent cells contributed significantly (P less than 0.001). Removal of adherent cells from peripheral blood mononuclear cells of patients and controls considerably increased in vitro lymphocyte responses to both mitogens and antigens (by about twice), suggesting that in this study, the inhibition of in vitro lymphocyte responses to antigens and mitogens by adherent cells was a general phenomenon independent of the presence of the disease.

Humans↗

PubMed1975

T-Lymphocyte deficiency following adult thymectomin man.

10 patients thymectomized about 20 years earlier were tested for delayed skin reactivity against mumps, streptokinase-streptodornase and PPD antigens. The T and B lymphocytes in peripheral blood were determined. 6 patients were surgically thymectomized and 2 of the received postoperative irradiation. 4 patients were treated with only high dose irradiation to the mediastinal compartment. None of the patients responded to all skin antigens, 1 was positive to two antigens and 3 to one antigen. The remaining patients were non-responders. The number of peripheral blood lymphocytes was slightly decreased in the patients. A significant decrease in the number of T-lymphocytes (p less than 0.005) was found in the whole patient group, and the depression was more pronounced in the surgically thymectomized patients (p less than 0.001). The number of B-lymphocytes was unaltered. It is concluded that the abnormalities in this patient group reflect a T-lymphocyte deficiency. The immunologic status of the patients before treatment is not known. However, in 3 patients with recently discovered untreated thymomas no abnormalities were found. Thus, adult thymectomy in man seem to lead to deficiency of cell mediated immunity, indicating an influence of the thymus in immunological functions during adult life.

Adult↗

PubMed1976

Cellular aspects of selective IgA deficiency.

Five patients with no detectable serum IgA (less than 20 mug/ml) and one patient with low serum IgA were compared to normal subjects. The number of circulating E-RFC was normal as was the lymphocyte DNA synthesis induced by PHA, Con A, and streptokinase-streptodornase. The patients had normal numbers of IgA-bearing lymphocytes and normal or increased numbers of B cells. Purified anti-immunoglobulin antibodies specific for IgG, IgA and IgM induced a normal lymphocyte DNA synthesis as did PWM. The patients' lymphocytes were able in vitro to transform into actively secreting IgA plasmocytes. This transformation was determined by counting the IgA and immunoglobulin-containing cells and then measuring the IgA and IgG secretion in the cultures. In some patients PWM was selectively suppressive in IgA B-cell transformation into IgA secreting cells; in the other patients PWM had no effect on the IgA B-cell differentiation. PWM enhanced the IgG secretion in the patients' cultures as well as IgA and IgG secretion in the normal controls.

Adult↗

PubMedDate not supplied

Immunotherapy of prostatic carcinoma with bacillus Calmette-Guerin.

Seventeen patients with histologically proven adenocarcinoma of the prostate were selected for evaluation of their immunologic competence and therapy with bacillus Calmette-Guérin (BCG). All patients were in stage D. The immune response was explored in two main aspects: cell-mediated and humoral immunity. Delayed skin hypersensitivity reaction with purified protein derivative (PPD), streptokinase-streptodornase (varidase), Candida, mumps antigen, and Trichophyton were tested. Lymphocyte reactivity was measured by in vitro blastogenesis. Serum immunoglobulin levels and serum protein electrophoresis were also measured. The patients were then divided in two groups according to the skin test response to PPD. Group 1 (PPD-positive) consisted of seven patients. Group 2 (PPD-negative) consisted of ten patients. In group 1, two patients were treated with intraprostatic injection of BCG every week in the following doses: 1 cc the first week, 2 cc the second week, 4 cc the third week, and 6 cc the fourth week. The five remaining patients in this group received only 1 cc every week for 4 weeks. The group 2 patients (PPD-negative) were stimulated by oral intake of BCG in an attempt to convert their skin tests to positive. All patients revealed varying degrees of immunodepression. None of the patients in group 2 (stimulated by oral intake of BCG) converted to positive skin tests. Three patients in group 1 treated with BCG showed necrosis of the tumor. The different aspects of immunodepression in this disease are analyzed and correlated to the clinical staging, histologic grading, and response to therapy. The mechanism of BCG action in advanced prostatic carcinoma is discussed.

Adenocarcinoma↗

PubMed1975

Immunity in tumours of the skin. Studies on cell-mediated immune reactions.

Cell-mediated immunity (CMI) was investigated in a series of patients with epithelial tumours of the skin and with Kaposi's sarcoma. In all 53 patients the peripheral blood lymphocyte count showed no substantial difference from the control values. The blasts count after phytohaemagglutinin stimulation in 30 patients ranged from 40-50% with no noticeable difference from the control values. Intradermal tests with the staphylococcal antigen, PPD-tuberculin and streptokinase-streptodornase carried out in 59 patients proved negative in a significant number of cases, mainly those of Kaposi's sarcoma. The non immune inflammatory response to chemical irritant (benzalkonium chloride) was more often negative (17 of 59 cases) than in controls (5 of 30 subjects). The DNCB sensitization test carried out in 28 patients was negative in 5 cases (2 cases of late basal-cell epithelioma, 2 cases of advanced squamous-cell carcinoma and 1 case of Kaposi's sarcoma). The findings do not thus indicate the presence of a gross CMI defect in the material studied, which would be borne out by all the tests employed. The sometimes discordant results of the various tests suggest the existence of partial CMI defects in certain tumours, but mainly in Kaposi's sarcoma. Such partial defects, encountered even in early neoplasms, coexist, in some cases, with failure of inflammatory response to chemical irritants.

Adult↗

PubMedDate not supplied

[Papillon-Lefèvre syndrome and recurrent infections].

Seven cases of Papillon-Lefevre syndrome are described, 4 children and their mother with two other relatives on the mother's side. An immune defect was suspected because six of the seven had a history of severe pyogenic infections, but the results of the tests were not the same in the three individuals tested. In one patient no immune defect was detected but in the other two the response of lymphocytes to candida and to streptokinase and streptodornase was reduced.

Adolescent↗

PubMedDate not supplied

Immune reactivity and clinical stage of patients with laryngeal cancer.

Selected parameters of the immune system were determined in vivo and vitro in 200 patients with malignant tumours of the larynx in various stages of clinical progression before surgical treatment. The comparison of the results with those in control groups of healthy persons made division of the patients possible into groups (a, b, c, d), and statistical analysis of the differences. The distribution of the patients according to the mean values of these parameters showed that such parameters as: white blood cell count, absolute lymphocyte count, absolute granulocyte count, receptor activity of T-cells and enzymatic activity of peripheral blood neutrophils may have characteristic values in laryngeal cancer. Particularly significant from the standpoint of prognostic value were the results of delayed hypersensitivity response to protein pure derivative (PPD), phytohaemagglutinin (PHA) and streptokinase-streptodornase (SK-SD).

Granulocytes↗

PubMed1991

[Delayed nonspecific cutaneous hypersensitivity in extrinsic allergic alveolitis and idiopathic pulmonary fibrosis].

BACKGROUND: A study of delayed nonspecific cutaneous hypersensitivity in extrinsic allergic alveolitis and in idiopathic pulmonary fibrosis. METHODS: 13 patients with extrinsic allergic alveolitis, 10 with idiopathic pulmonary fibrosis, 34 with sarcoidosis and a control group of 110 subjects without respiratory disease or apparent defects in cell immunity were evaluated. The skin tests of delayed hypersensitivity were carried out with five antigenic extracts (candidin, staphylococcal toxoid, tuberculin-PPD, trichophyllin, streptokinase-streptodornase). The reactions were read after 48 hours. Delayed nonspecific cutaneous hypersensitivity was globally calculated, both quantitatively (sum of the mean diameters) and quantitatively (overall average of positive reactions). RESULTS: No significant differences were found between the group of patients with idiopathic pulmonary fibrosis and the control group, nor between those with extrinsic allergic alveolitis and those with sarcoidosis. However, differences were found between the control group and the patients with extrinsic allergic alveolitis, both in the quantitative and the qualitative analysis. CONCLUSIONS: Delayed nonspecific cutaneous hypersensitivity in extrinsic allergic alveolitis is depressed in a similar way as in sarcoidosis, whereas it appears to be preserved in idiopathic pulmonary fibrosis. Therefore, extrinsic allergic alveolitis should be considered as another cause of reduction of delayed nonspecific cutaneous hypersensitivity as it is the case with sarcoidosis.

Adult↗

PubMed1979

Inhibition of human lymphocyte mitogen and antigen response by a 15,000-dalton protein from feline leukemia virus.

Peripheral blood lymphocyte response of normal human subjects to mitogens and antigens was suppressed by a 15,000-dalton protein (p15) from a C-type feline leukemia virus. Four of six subjects were suppressed 70 to 96% when responding to concanavalin A or phytohemagglutinin in the presence of 5.0 microgram of p15. The three subjects who responded to streptokinase-streptodornase and Candida were suppressed 68 to 91% when cultured with 5.0 microgram of p15. The subviral protein did not appear to be cytotoxic at doses reported. Lymphocyte membrane studies with fluorescein isothiocyanate-concanavalin A revealed a reduction in concanavalin A-induced cap formation of 51 to 91% in the presence of the same dose of p15. These results demonstrate that in vitro immunological dysfunction in human lymphocytes can be induced by a C-type virion protein.

Antigens↗

PubMed1990

A collaborative study to establish the second international standard for streptokinase.

An International Standard for Streptokinase--Streptodornase (62/7) has been used to calibrate high purity clinical batches of SK since 1965. An international collaborative study, involving six laboratories, was undertaken to replace this standard with a high purity standard for SK. Two candidate preparations (88/826 and 88/824) were compared by a clot lysis assay with the current standard (62/7). Potencies of 671 i.u. and 461 i.u. were established for preparations A (88/826) and B (88/824), respectively. Either preparation appeared suitable to serve as a standard for SK. However, each ampoule of preparation A (88/826) contains a more appropriate amount of SK activity for potency testing, and is therefore preferred. Accelerated degradation tests indicate that preparation A (88/826) is very stable. The high purity streptokinase preparation, coded 88/826, has been established by the World Health Organisation as the 2nd International Standard for Streptokinase, with an assigned potency of 700 i.u. per ampoule.

Drug Stability↗

PubMed1985

A histamine-releasing factor from activated human mononuclear cells.

Human mononuclear cells activated by streptokinase-streptodornase have been shown to elaborate a factor capable of releasing histamine from human basophils. We have developed reproducible methods for its production in large quantities by using cells obtained from leukapheresis packs, by detection utilizing donor basophils known to release well with anti IgE, and by quantitation of histamine by the radioenzyme method. Human histamine-releasing factor (HRF) gave a single peak upon gel filtration with an estimated m.w. of 32,000; SDS gel electrophoresis revealed a single major band as seen at m.w. 30,000. HRF can be resolved into at least two forms separable by ion-exchange chromatography on QAE Sephadex, and two peaks of activity were obtained by chromatofocusing or isoelectric focusing in gels at pH 6.9 and between 7.4 and 8.3. This factor represents an important potential link between cellular immunity and immediate hypersensitivity.

Basophils↗

PubMed1989

Effect of preincubation on the proliferative response to antigen by cells from leprosy patients and healthy controls.

One of the postulated mechanisms contributing to the selective T-cell hyporesponsiveness in patients with leprosy is receptor blockade, the characteristic feature of which is reversibility following preincubation of cells in vitro. To test this hypothesis, peripheral blood mononuclear cells (PBMC) obtained from 17 leprosy patients and 10 healthy Mantoux-positive and -negative controls were cultured freshly or after a period of preincubation in serum-containing medium, and the proliferative responses to Mycobacterium leprae, BCG, and streptokinase-streptodornase (SKSD) were measured. On the basis of the response to M. leprae, the leprosy patients could be divided into low, intermediate and high responders. Preincubation for 2-16 hr resulted in enhanced proliferation by cells from moderate responders, but not from low or high responders. Although the effect was serum dependent, it was neither antigen specific nor was it confined to cells from leprosy patients. Thus, an increase in response to both the crossreactive and unrelated antigens BCG and SKSD occurred, and the same trend was observed when cells from healthy controls were preincubated in serum-containing medium. Furthermore, cells from different individuals displayed varying responses to different antigens following preincubation, suggesting that the effect of this step was neither confined to isolated individuals nor to particular antigens. The addition of pronase to the preincubation step did not further enhance the response to antigen over and above that obtained with preincubation alone. It was therefore concluded that the enhanced proliferative response to antigen following preincubation was an in vitro phenomenon dependent upon the culture conditions employed, was not specific to leprosy, and was not related to receptor blockade.

Antigens, Bacterial↗