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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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

[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↗

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↗

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↗

Some immunological parameters in rheumatoid arthritis from India.

The present study describes the immune status of 41 patients with rheumatoid arthritis from India. The results showed a high level of immunoglobulins, normal levels of C3, less proportion of seropositivity (54%) than that reported from the west and 26% having antinuclear antibody in their serum. T lymphocyte proportion in the blood was found to be low in acutely ill patients only. Mantoux and streptokinase-streptodornase skin tests for delayed hypersensitivity were positive only in about 50% of that of controls. However, all the patients could be sensitized to DNCB. For testing the humoral antibody response TAB vaccine was given. The levels of antibody produced against O and H antigens of "Salmonella typhi" were comparable in patients and controls. However, the level of IgM rose significantly in the patients in comparison to that of controls. The cause of this, at least partially, was found to be due to a very significant rise of rheumatoid factor after TAB vaccine in the patients. Also 3 out of 8 patients converted to a seropositive state after TAB vaccination. Controls did not show the development of rheumatoid factor.

Agglutinins↗

Differential ability of fixed antigen-presenting cells to stimulate nominal antigen-reactive and alloreactive T4 lymphocytes.

The capacity of paraformaldehyde-fixed human antigen-presenting cells (APC) to induce responses by autologous, freshly isolated peripheral blood T4 cells was examined and was compared with their ability to stimulate allogeneic T4 cell DNA synthesis. Fixation of glass-adherent cells (AC) with as little as 0.06% paraformaldehyde abolished leucine incorporation, whereas fixation with 0.75% paraformaldehyde caused death of greater than 98% of the AC. Control APC were able to take up and present the soluble antigens streptokinase-streptodornase (SK-SD), tetanus toxoid, or tuberculin-purified protein derivative to autologous Ia-depleted T4 cells. Fixation with greater than 0.06% paraformaldehyde eliminated such ability. When AC were incubated with antigen overnight and were then fixed, however, they were able to present nominal antigen to autologous T4 cells in a genetically restricted manner that was blocked by monoclonal antibodies directed against monomorphic determinants on class II major histocompatibility complex (MHC) molecules. Despite the ability to present nominal antigen, paraformaldehyde-fixed AC were unable to induce allogeneic T4 cell proliferation. Similar results were observed when non-T cells or spleen cells were used as stimulators. The inability of fixed APC to stimulate allogeneic T4 cell DNA synthesis was not reversed by increasing the number of fixed APC or by the addition of control AC autologous to the responding cells. Moreover, interleukins 1 and 2 either alone or in combination also failed to permit maximal T cell proliferation in response to fixed allogeneic APC. The differential effects of fixation on nominal antigen and alloantigen presentation could not be explained by the loss of membrane thymocyte stimulatory activity on fixed AC. These results indicate that antigen-bearing fixed APC are competent to stimulate proliferation by antigen-reactive T4 cells, but are deficient at inducing allogeneic T4 cell DNA synthesis. The differential sensitivity of these two Ia-restricted functions of APC to chemical denaturation (reductive methylation) by paraformaldehyde suggests that the allodeterminants and restriction elements for nominal antigen on MHC class II molecules can be functionally dissociated.

Antigen-Presenting Cells↗

Dissection of defective antigen presentation by interferon-gamma-treated fibroblasts.

The capacity of interferon-gamma (IFN-gamma)-treated HLA-DR expressing human dermal fibroblasts (FB) to function as antigen-presenting cells (APC) was examined. FB were cultured with 250 U/ml IFN-gamma for 4 days to induce HLA-DR expression. Peripheral blood monocytes (M phi), FB, or IFN-gamma-treated FB from the same donor were then cultured overnight with or without the recall antigen streptokinase streptodornase (SKSD), and their capacity to stimulate autologous T4 cell DNA synthesis was examined. SKSD-bearing M phi stimulated T4 cell proliferation, whereas antigen-bearing HLA-DR (+) FB did not. Even after fixation with paraformaldehyde to eliminate metabolic activity, SKSD-bearing M phi, but not FB, were able to function as APC. However, when HLA-DR (-) endothelial cell (EC) or autologous or HLA-D-mismatched M phi were added to the cultures, antigen-pulsed IFN-gamma-treated FB and M phi were comparably effective stimulators of autologous T4 cell DNA synthesis. Antigen recognition by the T4 cell was restricted by the class II major histocompatibility complex (MHC)-encoded gene products expressed by the IFN-gamma-treated FB and was unrelated to the class I or II MHC-encoded gene products expressed by the additional cell type. EC-promoted T4 cell DNA synthesis induced by antigen-bearing IFN-gamma-treated FB was inhibited by 60.3, a monoclonal antibody directed at an epitope common to LFA-1, CR3, and the p150,95 molecule. Inhibition caused by 60.3 was completely reversed by the addition of IL 2 to the cultures. Antigen presentation by IFN-gamma-treated FB was also enhanced somewhat by IL 1, IL 2, or monoclonal antibody directed at Tp44 (9.3). However, each of these additions alone promoted T cell proliferation less effectively than EC and resulted in responses that were smaller than those triggered by antigen-bearing M phi. The data suggest that IFN-gamma-treated FB take up and process antigen effectively, but lack an accessory cell property necessary for antigen-induced T4 cell IL 2 production and proliferation.

Antigen-Presenting Cells↗

Cell-mediated immunity in vitro in atopic dermatitis.

The function of T cells in atopic dermatitis was studied by leukocyte migration agarose and lymphocyte transformation tests. We found that phytohemagglutinin (PHA)- and PPD-induced release of leukocyte migration inhibition factor (LIF) from lymphocytes is significantly decreased (P less than .001) in patients with atopic dermatitis as compared with healthy controls. Blastogenic response of lymphocytes induced by PPD was also decreased in atopic patients as compared with controls (P less than .01). No differences were found in spontaneous blastogenesis or in blastogenic response of lymphocytes in vitro to PHA, concanavalin A (ConA), or streptokinase-streptodornase (SK-SD) between patients with atopic dermatitis and controls.

Adolescent↗

Immunology of atopic dermatitis.

Thirty-two adult patients with atopic dermatitis were studied at the Allergology Service of the "Hnos. Ameijeiras" Clinical Surgical Hospital. The diagnosis was established following the criteria of Hanifin and Lobitz. A detailed medical history was written for the patients; the study of some immunological parameters, such as the serum immunoglobulin quantification, delayed skin tests with a battery of antigens, and the spontaneous rosette-test, was also carried out. Almost all the patients showed serum IgE values above 150 UI, by means of the ELISA test modified by C.E.N.I.C. The mean values of the spontaneous rosette-test were low; this was more noticeable during the exacerbation period of the lesions. Candida sp, Mantoux and Streptokinase-Streptodornase antigens showed negative results in a high proportion of patients with atopic dermatitis, in relation with the control group. In atopic dermatitis, there are humoral disorders of immunity; this was demonstrated in our group by increased values of IgE and cellular disorders due to skin anergy, and to a low percentage of rosette forming cells; this does not allow to state that these phenomena have an active participation in the etiopathogenesis of this entity.

Adult↗