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PubMed2003

Prophage induction and expression of prophage-encoded virulence factors in group A Streptococcus serotype M3 strain MGAS315.

The genome of the highly virulent group A Streptococcus (GAS) serotype M3 strain MGAS315 has six prophages that encode six proven or putative virulence factors. We examined prophage induction and expression of prophage-encoded virulence factors by this strain under in vitro conditions inferred to approximate in vivo conditions. Coculture of strain MGAS315 with Detroit 562 (D562) human epithelial pharyngeal cells induced the prophage encoding streptococcal pyrogenic exotoxin K (SpeK) and extracellular phospholipase A(2) (Sla) and the prophage encoding streptodornase (Sdn). Increased gene copy numbers after induction correlated with increased speK, sla, and sdn transcript levels. Although speK and sla are located contiguously in prophage Phi315.4, these genes were transcribed independently. Whereas production of immunoreactive SpeK was either absent or minimal during coculture of GAS with D562 cells, production of immunoreactive Sla increased substantially. In contrast, despite a lack of induction of the prophage encoding speA during coculture of GAS with D562 cells, the speA transcript level and production of immunoreactive streptococcal pyrogenic exotoxin A (SpeA) increased. Exposure of strain MGAS315 to hydrogen peroxide, an oxidative stressor, induced the prophage encoding mitogenic factor 4 (MF4), and there was a concomitant increase in the mf4 transcript. All prophages of strain MGAS315 that encode virulence factors were induced during culture with mitomycin C, a DNA-damaging agent. However, the virulence factor gene transcript levels and production of the encoded proteins decreased after mitomycin C treatment. Taken together, the results indicate that a complex relationship exists among environmental culture conditions, prophage induction, and production of prophage-encoded virulence factors.

Bacterial Proteins↗

PubMed1982

In vitro lymphocyte proliferation in response to type III group B streptococci.

The in vitro cell-mediated responses to group B streptococci (GBS) and the relationship of cell-mediated immunity to specific humoral immunity to type III GBS were investigated. Blood was obtained from 20 adult volunteers, and lymphocytes were isolated and cultured in microtiter plates. Each well contained 2 x 10(5) lymphocytes, 15% autologous serum, and either GBS (cell-to-organism ratio of 1:10, 1:1, or 1:0.1), phytohemagglutinin, streptokinase-streptodornase, or RPMI 1640. Cells were harvested at 5, 6, or 7 days, and DNA synthesis was quantitated. Serum antibody titers were determined with an enzyme-linked immunosorbent assay. Maximal lymphocyte responses occurred at 6 days of culture and at a cell-to-organism ratio of 1:1. Individuals with significant antibody titers to type III GBS, as well as those with undetectable antibody, responded to GBS (stimulation index greater than 10). There was a significant difference (P less than 0.001) between mean antibody concentrations in responders (stimulation index greater than 10) and nonresponders (stimulation index less than 10). Thus, the in vitro responses to GBS may be both to a specific antigen and to a nonspecific mitogen and may be important in host immunity to GBS.

Adult↗

PubMed1979

Treatment of rheumatoid arthritis with levamisole: long-term results and immune changes.

We treated 29 rheumatoid arthritis patients with levamisole. on the basis of a 25% improvement in any 3 of 6 measurements 95% of the patients had a favourable response within 20 weeks. However, 64% of the patients discontinued levamisole by 40 to 60 weeks because of rash or secondary treatment failures. Delayed skin reactivity to streptokinase-streptodornase increased significantly in the entire treatment group, but there was in inverse correlation between skin test enhancement and clinical response. There was no overall change in lymphocytes response to phytohaemagglutinin (PHA) after 4 and 16 weeks of treatment, but seven patients with enhanced lymphocyte responsiveness to PHA experienced an earlier clinical response to levamisole. Treatment with levamisole frequently results in clinical improvement in rheumatoid arthritis, but this is not clearly related to a stimulatory effect on cell-mediated immunity. Its long-term usefulness may be limited by a high incidence of relapse and rash.

Arthritis, Rheumatoid↗

PubMed1983

Depressed lymphocyte transformation by yersinia and Escherichia coli in yersinia arthritis.

Lymphocyte transformation responses were studied in 21 patients with acute yersinia infection followed-up after the acute infection for up to one year. Eight patients had reactive arthritis caused by yersinia. The responses to the causative serotype of yersinia were significantly lower (p less than 0.05) in patients with arthritis than in those without arthritis. Stimulation with Escherichia coli gave lower responses than with yersinia, but with E. coli the difference between arthritic and nonarthritic groups was more significant (p less than 0.02). The responses to yersinia and E. coli were not correlated with the presence of HLA B27. Lymphocyte transformation by purified protein derivative of tuberculin, streptokinase-streptodornase, phytohaemagglutinin, or concanavalin-A revealed no significant differences between the arthritic and nonarthritic groups. The role of the enterobacterial common antigen in the pathogenesis of reactive arthritis and ankylosing spondylitis is discussed.

Adolescent↗

PubMed1989

Impaired non-specific delayed cutaneous hypersensitivity in bird fancier's lung.

The relation between non-specific delayed cutaneous hypersensitivity and bird fancier's lung was investigated in 13 patients with the disorder. They were compared with 50 subjects who had no reason to have decreased non-specific delayed cutaneous hypersensitivity (control group) and 34 patients with pulmonary sarcoidosis. In addition, 13 patients with bird fancier's lung (11 of the original group) were tested at least one year after avoiding exposure to the causal antigen. Five antigens (candidine, staphylococcal toxoid, tuberculin purified protein derivative, trichophyton, and streptokinase-streptodornase) were injected intradermally (0.1 ml) and the mean weal diameter was measured at 48 hours. The mean weal size was significantly less in the subjects with bird fancier's lung at the time of diagnosis than in the control group (2.23 v 5.66 mm) but did not differ significantly from that of the subjects with sarcoidosis (2.80 mm) or from that of the bird fanciers with no exposure to the causal antigen for one year (2.75 mm). The impairment of non-specific delayed cutaneous hypersensitivity in patients with bird fancier's lung appears to be quantitatively similar to that occurring in sarcoidosis.

Adult↗

PubMed1981

Orally and rectally administered streptokinase. Investigation of its absorption and activity.

The absorption and activity of orally and rectally administered streptokinase was investigated. No difference was found between the levels of plasminogen activators and antistreptokinase antibodies before and after oral or rectal administration of streptokinase-streptodornase (Varidase), nor was there any difference in the hematoma resorption times between patients treated with oral streptokinase and a control group. We found no evidence of streptokinase absorption or increased fibrinolytic activity in plasma after oral or rectal administration in our patients.

Administration, Oral↗

PubMed1977

Effect of blood transfusion on the immune response of children with thalassaemia.

The effect of blood transfusions on the immune response of 46 thalassaemic children was studied. Cell-mediated immune response was evaluated by performing skin tests to specific (streptokinase-streptodornase and candidin) and nonspecific (dinitrochlorobenzene and phytohaemagglutinin) antigens. Antibody response to specific antigen (tetanus toxoid) was estimated by measuring the tetanus antitoxin titre before and after vaccination. No gross impairment of cell-mediated immunity was elicited. The larger proportion of negative phytohaemagglutinin skin tests found in thalassaemic patients does not necessarily suggest a cell-mediated immunity impairment, since this skin reaction is also affected by other factors, especially the inflammatory skin response. The transfused antibodies may inhibit the recipient's sensitization and primary immune response to the homologous antigen, especially when the antibody level in the transfused blood is high whereas the secondary immune response is not affected.

Adolescent↗

PubMed1977

Lymphocyte reactivity in healthy subjects and cancer patients.

Lymphocytes of 40 cancer patients with solid tumors as well as of 40 normal subjects were stimulated by phytohemagglutinin (PHA), purified protein derivative (PPD), streptokinase-streptodornase (SK-SD) and candidin (Cand) and the extent of their stimulation was expressed by the index of reactivity. The cumulative frequency distribution of reactivity indices was significantly higher in normal individuals than in cancer patients. The highest extent of information relevant to the discrimination between patients and normal subjects is obtained following stimulation of lymphocytes with Cand and in decreasing order with PPD, SK-SD and PHA. The percentage of normal subjects and patients having reactive lymphocytes was found to be the highest following stimulation with PHA, lower with PPD and the lowest to Cand and SK-SD. The importance of using increasing numbers of stimulants to achieve maximal detection of lymphocyte reactivity is documented and discussed.

Adult↗

PubMed1979

Cell-mediated immunity in patients with cervical cancer.

In 16 patients with cervical carcinoma (stages II and III) tumor-specific and general immune reactivity was investigated. Cancer patients' skin reactivity was different from that of controls insofar as there was a decreased response to streptokinase-streptodornase as well as to DNCB. Absence of DNCB sensitization was mainly in stage III patients. Cell-mediated immunity to autologous and/or allogeneic tumor-associated antigens (TAA) was demonstrated in vivo and in vitro. 3 of 8 patients had positive skin reactions to autologous TAA and 8 of 14 reacted to allogeneic TAA. In the lymphocyte migration inhibition test, 8 of 14 patients reacted to autologous tumor membrane preparations and 3 of 10 to allogeneic pooled preparations. In addition, there was some indication of cross-reaction with antigens from a human squamous cell carcinoma of the lung. There was no reaction in vitro to normal cervical tissue and no association between tumor-specific reactions and herpes simplex virus type II antibodies could be demonstrated. Patients with DNCB sensitization and in vivo as well as in vitro lymphocyte reactivity to TAA had a better prognosis than nonreacting patients, indicating the value of combined immunological tests.

Adult↗

PubMed1976

Discrepancy between skin reactivity and blastogenic activity of fractionated PHA-P.

Firstly, it was reported that phytohemagglutinin P (PHA-P) can induce the immediate skin reaction besides the delayed one. 70 out of 80 cases (87.5%) gave positive immediate reactions with 1 mug of PHA-P. There was a close correlation between the radioallergosorbent test (RAST) counts against PHA-P and the total IgE value or erythematous diameter of the immediate skin reactions. Absorption with anti-IgE or PHA-P reduced the RAST counts against PHA-P to 79 and 56% of their initial values, respectively, but did not effect the RAST counts against steptokinase-streptodornase (SK-SD). These results suggest that the immediate skin reaction might be induced by specific IgE antibody against PHA-P. Secondly, using three fractions of PHA-P obtained by Sephadex G-200 gel filtration, skin test and RAST in humans, ACA in rats, and 3H-thymidine incorporation into human leukocytes, were carried out and data obtained that showed some discrepancy between them. That is, on skin testing, it was shown that all of the three fractions (fr.) had the ability to elicit the immediate and delayed skin reaction in humans and rats with the order of fr. 1, 2 and 3, and it was confirmed by RAST. On the other hand, the leukocyte-stimulating activity resided in fr. 1 and 2, while fr. 3, which seemed to be a group of subunits with a molecular weight of around 35,000, had almost no activity. These findings have been discussed in view of the earlier studies on the structure of PHA-P

Absorption↗

PubMed1984

The value of the DNCB test in bladder cancer. Pretreatment evaluation of immune function and 5-year follow-up of patients with urinary bladder cancer.

In 68 patients with histologically verified tumors of the urinary bladder, cell-mediated and humoral immune parameters were investigated before therapy and the results were re-evaluated after a 5-year observation period in order to correlate them with relapse rate and survival time. Skin test reactivity, as measured with recall antigens (tuberculin, streptokinase-streptodornase, mumps, toxoplasmin and candidin), and serum levels of immunoglobulins do not differentiate between levels of invasion and grade of malignancy. However, it was found that patients with tumors of high grades of invasiveness and malignancy were anergic to the primary skin test antigen dinitrochlorobenzene (DNCB). Furthermore, a correlation between anergic reactivity to the DNCB test and absence of local inflammatory reactions at the tumor site was detected, showing that patients with a negative DNCB challenge test were those in whom no immunocytes could be detected in the intra- and peritumoral area. Survival time and incidence of relapse were also correlated with initial skin test reactivity to DNCB, i.e. all patients with tumor stage pT3 and skin test anergy developed recurrences and died within the 5-year observation period. The correlation between morphological inflammatory criteria and immunological parameters detected in patients with advanced tumor stages should therefore be taken into consideration when taking therapeutic decisions at the time of diagnosis.

Aged↗

PubMed1977

Cell-mediated immunity in silicosis.

Selected parameters of cell-mediated immunity were determined in a group of 16 patients with silicosis. The results were compared with those of a control group of 13 subjects without silica exposure. There were no group differences in the mean number of delayed hypersensitivity skin tests to a battery of recall antigens (purified protein derivative, candida, streptokinase-streptodornase, and trichophyton) or in the mean number of peripheral blood lymphocytes. Lymphocyte responsiveness to phytohemagglutinin, pokeweed mitogen, and the antigens listed, and the percentage of lymphocytes that formed sheep red blood cell rosettes (t cells) and complement rosettes (B cells) were also similar in both groups; however, the silica-exposed group demonstrated depressed lymphocyte stimulation in response to low concentrations of concanavalin A.

Humans↗

PubMed1986

Human alveolar macrophage regulation of lymphocyte proliferation.

Human alveolar macrophages obtained by bronchoalveolar lavage have different accessory cell characteristics than do autologous, peripheral-blood-derived macrophages for the support of lymphocyte proliferative responses. To further investigate these differences, we performed mixing experiments in which variable numbers of the 2 macrophages were cocultured and assayed for support of mitogen-(phytohemagglutinin; PHA) and antigen-(streptokinase-streptodornase; SK-SD) stimulated proliferation of purified autologous lymphocytes. Alveolar macrophages promoted greater lymphocyte proliferation, in proportion to their added number in coculture with peripheral-blood-derived macrophages, when using a suboptimal concentration of PHA. In contrast, even small numbers of alveolar macrophages suppressed lymphocyte proliferation supported by peripheral-blood-derived macrophages when using the optimal concentration of PHA or the antigen SK-SD. The data indicated that diminished support exhibited by alveolar macrophages for optimal mitogen-stimulated or antigen-stimulated lymphocyte responses was due to active suppression by the cells, rather than to deficient or defective accessory cell function. Thus, alveolar macrophages have regulatory effects on lymphocyte proliferation that are dependent on type and concentration of mitogenic stimulation. Such regulation may relate to alveolar macrophage function in vivo, providing appropriate response yet limiting inflammation in the lung.

Adult↗

PubMed1972

Effect of transfer factor on lymphocyte function in anergic patients.

Dialyzable transfer factor, obtained from frozen-thawed peripheral blood leukocytes from a single donor, was given to five anergic patients with chronic mucocutaneous candidiasis. Studies of immunological responses including delayed cutaneous hypersensitivity, in vitro antigen-induced thymidine incorporation, and production of macrophage migration inhibition factor (MIF) were conducted both before and after injection of transfer factor. Before transfer factor, none of the patients had delayed skin responses to any of the natural antigens studied. Their lymphocytes did not produce MIF after exposure to antigens in vitro and only one patient showed increased thymidine incorporation when his lymphocytes were cultured with candida and streptokinase-streptodornase (SK-SD). After injection of transfer factor, four patients developed delayed skin responses to antigens to which the donor was sensitive; no recipient reacted to an antigen to which the donor was nonreactive. Lymphocytes from recipients produced MIF when cultured with antigens that evoked positive delayed skin tests. Only one patient developed antigen-induced lymphocyte transformation and this response occurred only intermittently. Attempts to sensitize three of the patients with the contact allergen, chlorodinitrobenzene, both before and after transfer factor, were unsuccessful. The fifth patient, a 9-yr old boy with an immunologic profile similar to the Nezelof syndrome, did not become skin test-reactive or develop positive responses to the in vitro tests. These findings suggest that transfer factor acts on the immunocompetent cells that respond to antigens with lymphokine production, but has little, if any, effect on cells that respond to antigens by blastogenesis. The failure to sensitize the subjects with chlorodinitrobenzene illustrates the specificity of the immunologic effects of transfer factor, and implies that it does not function through nonspecific, adjuvant-like mechanisms. Failure of transfer factor to produce positive skin tests or MIF production in a patient with Nezelof's syndrome may be evidence that lymphokine-producing cells are thymus derived.

Adult↗

PubMed1974

Serial studies of immunocompetence of patients undergoing chemotherapy for acute leukemia.

Immunocompetence was followed serially for 1 yr from the onset of treatment in 55 adult patients with acute leukemia. The tests used were delayed hypersensitivity responses to a battery of five recall antigens (dermatophytin, dermatophytin 0, candida, streptokinase-streptodornase, and mumps) and in vitro lymphocyte blastogenic responses to phytohemagglutinin and streptolysin 0. There was a strong correlation between immunocompetence at the start of treatment and a good prognosis; 32/39 patients who subsequently entered remission were initially immunocompetent compared to 4/15 who failed to enter remission. In the complete remission group there was a decline in competence starting from 2 to 5 mo after the onset of treatment. In those who remained in remission for 1 yr, competence recovered at 6 mo and remained vigorous thereafter. In those who relapsed before 1 yr, the decline in competence occurred 1 mo before relapse and competence continued to decline progressively during the 1 yr follow-up period. These studies suggest that therapeutic approaches which restore immunocompetence or prevent its decline will improve both the remission rate and the remission duration of patients with acute leukemia.

Adolescent↗

PubMed1977

Studies on the pathogenesis of an immune defect in multiple myeloma.

The reduced capacity of patients with multiple myeloma to respond to antigen challenge is well recognized. Response to antigen involves antigen recognition, cell proliferation, and synthesis and secretion of antibody. This study examines this sequence of events in peripheral blood lymphocytes from untreated and treated patients with myeloma, from individuals with benign monoclonal gammopathy, and from normal healthy donors. Antigen-binding capacity was assessed by testing the ability of lymphocytes to bind radio-labeled pneumococcal polysaccharide, tetanus toxoid, or diptheria toxin. The in vitro proliferative response to these antigens as well as to pokeweed mitogen and streptokinase-streptodornase was evaluated. The secretion of immunoglobulin in response to pneumococcal polysaccharide, tetanus toxoid, and pokeweed mitogen by 2-4 x 10(6) lymphocytes in 7-day cultures was determined. The effects of coculture of myeloma peripheral blood lymphocytes and normal peripheral blood lymphocytes on immunoglobulin production and mixed leukocyte reactions were explored. All myeloma patients had normal numbers (3-8/5,000 cells) of antigen-binding cells. However, most showed a diminished antigen-induced blast transformation as measured by uptake of [(125)I]5-iodo-2'-deoxyuridine in culture. Immunoglobulin production in response to specific antigen in myeloma lymphocytes was 30-80% less than in normal lymphocytes. Immunoglobulin synthesis and mixed leukocyte responses by normal peripheral blood lymphocytes could be suppressed by myeloma lymphocytes. Multiple suppressor populations were present. Thus, the immune defect in myeloma is beyond the antigen recognition step and involves both the proliferation of antigen-sensitive cells and immunoglobulin production. Further suppressive effects are imposed on normal cells, implying defects in immunoregulation in this disease.

Adult↗

PubMed1979

Delayed-type hypersensitivity skin reactions in congenital afibrinogenemia lack fibrin deposition and induration.

Induration is a characteristic feature of delayed-type hypersensitivity skin reactions and is the usual measure of their intensity. The precise basis of induration has not been established, although activation of the clotting system with consequent fibrin deposition has been clearly implicated. In this study, two subjects with congenital afibrinogenemia, a genetic defect in fibrinogen synthesis, were skin tested with standard microbial antigens: streptokinase-streptodornase, monilia, mumps, and tuberculin purified protein derivative. One positive delayed reaction from each subject was biopsied at 40-48 h and compared with 23 biopsies of similar skin tests in normal volunteers. The eight skin tests in the afibrinogenic subjects lacked induration, although the erythema was similar in size (10-34 mm in diameter), intensity, and time-course to those in normals. Biopsies from the two strongest reactions from the afibrinogenemic subjects showed a typical perivascular mononuclear infiltrate. No more than traces of fibrin/fibrinogen were detected by immunofluorescence, in striking contrast to the abundant fibrin/fibrinogen deposition in 23 positive, indurated reactions in normal subjects. These findings indicate that fibrinogen itself is essential for the development of induration in delayed-type skin reactions in man. As judged by 1-mum sections and fluorescence, this is probably a result of the formation of an extravascular fibrin gel.

Adult↗

PubMed1984

Modulation of the immune response by gangliosides. Inhibition of adherent monocyte accessory function in vitro.

Gangliosides are potent inhibitors of lymphoproliferative responses. Selectively greater inhibitory effects of gangliosides on antigen-induced (vs. mitogen-induced) proliferation have been documented; e.g., 50 nmol of highly purified bovine brain gangliosides (BBG)/ml caused greater than or equal to 87% inhibition of proliferative responses of human peripheral blood mononuclear cells (PBMC) to three soluble specific antigens (Candida, streptokinase-streptodornase, and tetanus toxoid) vs. less than or equal to 37% inhibition of responses to three nonspecific mitogens (phytohemagglutinin, concanavalin A, and pokeweed mitogen). The possibility that BBG interfere with adherent monocyte accessory function, upon which responses to soluble specific antigens are strictly dependent, was therefore considered. PBMC were separated into the adherent and nonadherent subpopulations, exposed to BBG, recombined, and their proliferative responses were measured. Unseparated PBMC preincubated for 48-72 h with 100 nmol BBG/ml and then washed to remove unbound BBG exhibited 73-76% inhibition of subsequent antigen-induced lymphoproliferation. Separate pretreatment of both adherent and nonadherent cell subpopulations in BBG under the same conditions resulted in similar (72-82%) inhibition, which was reproduced by preincubation of only the adherent cells in BBG. Preincubation of only the nonadherent cells in BBG was not inhibitory. Inhibition (a) was independent of whether gangliosides were added in solution or incorporated into liposomes, (b) was abrogated by adding untreated monocytes to cultures containing adherent cells that were preexposed to BBG (excluding the possibility that BBG was inducing suppression mediated by adherent cells), and (c) was reversible by further incubation of BBG-pretreated adherent cells in control medium. Together, these results delineate a mechanism by which gangliosides modulate lymphoproliferative responses--direct, noncytotoxic, and ultimately reversible inhibition of the accessory function of adherent monocytes.

Animals↗