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Cell-mediated immunity in diabetes mellitus.

Cell-mediated immunity was evaluated in patients with diabetes mellitus by delayed hypersensitivity skin tests and in vitro lymphocyte transformations. Only 44% of diabetic patients had skin test reactivity to Candida antigen, compared with 88% of normal controls (P < 0.001). Insulin-dependent diabetic (IDD) patients had abnormally low lymphocyte transformation responses to phytohemagglutinin, concanavalin A, and streptokinase-streptodornase (P < 0.05). This defect was not corrected by culturing the cells in nondiabetic plasma. IDD patients with persistent hyperglycemia (fasting serum glucose level, >200 mg/dl) had lower levels of transformation than did IDD patients with fasting serum glucose levels less than 150 mg/dl. Lymphocytes from two IDD patients with poor lymphocyte transformation responses had marked improvement in response to phytohemagglutinin when the lymphocytes were cultured after a preincubation period designed to deplete cultures of suppressor activity. Seven IDD patients were studied serially over 12 months. Lymphocyte transformation responses in four of these patients improved coincidentally with a change in the level of fasting hyperglycemia from >200 to <150 mg/dl. The other three IDD patients with consistent fasting serum glucose levels of >200 mg/dl had poor lymphocyte transformation responses. Diabetic patients have demonstrable defects in lymphocyte function which improved in a small number of patients with reduction in the level of fasting hyperglycemia.

Adolescent↗

Modulation of immunoreactivity to periodontal disease-associated microorganisms during pregnancy.

The lymphocyte blastogenic response to a panel of antigens and mitogens was assessed in a group of 20 women throughout their pregnancy. In addition, a group of five nonpregnant women was monitored simultaneously to identify variations in response to the same stimulants. The stimulants included orally associated bacterial antigens (Streptococcus sanguis, Actinomyces viscosus, Bacteroides asaccharolyticus, Bacteroides melaninogenicus subsp. intermedius, Bacteroides [Capnocytophaga] ochraceus, and Fusobacterium nucleatum) and non-orally associated-stimulants (streptokinase-streptodornase, tetanus toxoid, concanavalin A, phytohemagglutinin, and pokeweed mitogen). Intrinsic (cells cultured in male AB plasma) suppression of the lymphocyte response to these stimulants was observed to occur by the second trmester of pregnancy and was resolved after parturition. Additionally, an extrinsic (cells cultured in autologous plasma) suppression was also suggested to occur in a similar manner. There was no detectable enhancement of the blastogenic response to oral bacteria associated with elevated gingivitis, which is generally reported to occur during nonpregnancy gingivitis. We propose that concomitant immunosuppression occurs during the second trimester, which masks such enhancement.

Actinomyces↗

In vitro correlates of delayed hypersensitivity in man: ambiguity of polymorphonuclear neutrophils as indicator cells in leukocyte migration test.

Delayed cutaneous hypersensitivity (DCH) of 12 normal adult subjects to purified protein derivative (PPD) of Mycobacterium tuberculosis, streptococcal streptokinase-streptodornase (SK-SD), and Candida albicans Dermatophytin O (DO) was assayed in vivo by skin testing and compared with such in vitro correlates of cellular immunity as lymphocyte transformation (LT) and inhibition of leukocyte migration (ILM) from microcapillary tubes or in agarose gel. LT was shown to be the best in vitro correlate of specific lymphocyte sensitization with all antigens. In the ILM assays, PPD showed good correlation with in vivo DCH and in vitro LT; SK-SD showed partial correlation; DO showed no correlation, not being active in any of the ILM tests. Cell distribution and morphology of stained migration patterns, ILM tests performed on separated populations of lymphocytes and polymorphonuclear leukocytes (PMN), as well as the ability of test antigens to stimulate PMN cells to reduce nitroblue-tetrazolium dye, indicated that in ILM tests mononuclear cells were not inhibited in their migration, whereas migration of PMN cells appeared to depend on their direct reaction with the test antigens.

Antigens, Bacterial↗

Cellular requirements for simulation by the purified streptococcal antigen DSR.

Delayed skin reactor (DSR) was prepared from streptokinase-streptodornase. Peripheral blood mononuclear cells, were collected from a Ficoll-Hypaque gradient and subsequently subdivided into T cell-, B cell-, or monocyte-depleted populations. Mononuclear cells with complement receptors (B cells) were removed by a nylon-wool column adsorption. Sheep erythrocyte rosette forming cells (T cells) were removed by centrifugation on a second gradient. Populations of cells which did not adhere to plastic and/or ingest iron filings were monocyte-depleted. While response to DSR as measured by incorporation of tritiated thymidine into DNA was not affected by removing B lymphocytes, it was significantly diminished by removing T lymphocytes. Monocyte-depleted cells incorporated less tritiated thymidine into DNA in both antigen stimulated and control cells, but the stimulation index was not significantly altered.

Antigens↗

Biologic activity of transfer factor tested in vitro with cord blood lymphocytes.

Cord blood lymphocytes, generally believed to be immunologically uncommitted, were used for investigating the possible conversion by transfer factor (TF) of non-immune to immune lymphocytes in vitro. TF was prepared from pooled buffy coats of normal blood bank donors by ultrafiltration and lyophilization, and it ability to influence lymphocyte transformation was assessed by measuring increases in 3H-thymidine uptake into DNA after stimulation with purified protein derivative of tuberculin and/or streptokinase-streptodornase in 23 experiments with cord blood and 15 adult controls. TF stimulated cord blood lymphocytes nonspecifically when added to the medium alone. In the presence of antigen, TF acted in a synergistic, antigen-dependent way with either amplifying or inhibitory effects in adult good responders. These effects were negligible in adult low responders and in cord blood. From our observations no specific conversion of lymphocytes by TF in vitro can be deduced.

Adult↗

Alveolar macrophage accessory cell function in bronchial asthma.

The capacity of peripheral blood monocytes and alveolar macrophages (AM) obtained by bronchoalveolar lavage (BAL) to present recall antigens, namely, tuberculin purified protein derivative (PPD) or streptokinase-streptodornase (SKSD), to highly purified autologous T-cells has been studied in 11 asthmatic and 11 healthy, nonatopic normal subjects. In the asthmatic group, AM accessory cell function was variable, and most subjects were unable to present either recall antigen as effectively as blood monocytes, although one asthmatic subject demonstrated larger proliferative responses than blood monocytes for both antigens. AM accessory cell activity was not antigen-specific, and there was a correlation between accessory cell efficacy for the two antigens (r = 0.92; confidence interval, 0.53 to 0.98). Furthermore, a correlation existed between the percentage lymphocyte count in the BAL fluid and the ratio of macrophage to monocyte antigen-presenting capability for both PPD (r = 0.92; 95% confidence interval, 0.83 to 0.99) and SKSD (r = 0.90; 95% confidence interval, 0.45 to 0.98). In the normal subjects, AM were also unable to act effectively as accessory cells for the presentation of PPD and SKSD in the majority of subjects. No correlation existed between the percentage lymphocytes in BAL fluid and the ratio of AM to monocyte accessory cell function. These results suggest an association between AM accessory function and the presence of BAL lymphocytes in bronchial asthma.

Adult↗

Factors of delayed hypersensitity in pulmonary tuberculosis.

In a study of delayed hypersensitivity in 54 patients with pulmonary tuberculosis who were on specific treatment, 33 patients could not be sensitized to dinitrochlorobenzene (DNCB) although 25 of these were tuberculin (PPD) positive. The majority of the patients also had positive skin reactions to another recall antigen, streptokinase-streptodornase. To exclude a drug effect on DNCB reactivity, 40 untreated patients were studied. Twenty-six (65 per cent) of these did not show DNCB reactivity. The cause for nonsensitization to DNCB was not clear. There was no abnormality in the number of T lymphocytes in the peripheral blood of 28 patients.

Adolescent↗

The effect of in vivo hydrocortisone on subpopulations of human lymphocytes.

This study was designed to determine the effect of in vivo hydrocortisone on subpopulations of lymphoid cells in normal humans. Subjects received a single intravenous dose of either 100 mg or 400 mg of hydrocortisone, and blood was drawn at hourly intervals for 6 h, and then again at 10 and 24 h after injection. Profound decreases in absolute numbers of circulating lymphocytes and monocytes occurred at 4-6 h after both 100 mg and 400 mg of hydrocortisone. Counts returned to normal by 24 h. The relative proportion of circulating thymus-derived lymphocytes as measured by the sheep red blood cell rosette assay decreased maximally by 4 h and returned to base line 24 h after hydrocortisone. There was a selective depletion of functional subpopulations of lymphocytes as represented by differential effects on in vitro stimulation with various mitogens and antigens. Phytohaemagglutinin response was relatively unaffected, while responses to concanavalin A were significantly diminished. Responses to pokeweed mitogen were unaffected by 100 mg of hydrocortisone, but greatly diminished by 400 mg of hydrocortisone. In vitro responses to the antigens streptokinase-streptodornase and tetanus toxoid were markedly diminished by in vivo hydrocortisone. Reconstitution of monocyte-depleted cultures with autologous monocytes partially corrected the diminished response to antigens. This transient selective depletion of monocytes and subsets of human lymphocytes by a single dose of hydrocortisone is most compatible with a redistribution of these cells out of the circulation into other body compartments.

Adult↗

Depression of the lymphocyte transformation response to microbial antigens and to phytohemagglutinin during pregnancy.

Lymphocyte transformation (LT) responses to Chlamydia trachomatis, to four other microbial antigens, and to phytohemagglutinin (PHA) were studied in 201 women during pregnancy and/or 3-18 wk postpartum. The LT responses to all stimulants tested were significantly depressed during pregnancy when compared with postpartum LT responses. This difference occurred whether LT assays were performed in autologous or pooled heterologous plasma collected from nonpregnant donors. Among women studied in the third trimester and again postpartum, the autologous LT stimulation index (LTSI) rose from 1.7 to 3.4 (P less than 0.001) with C. trachomatis elementary body antigen, from 3.7 to 7.9 (P less than 0.001) with Candida albicans cell wall extract, from 4.5 to 7.8 (P = 0.008) with streptokinase-streptodornase, from 1.7 to 3.0 (P = 0.007) with fluid tetanus toxoid, from 1.7 to 2.8 (P = 0.046) with mumps virus skin test antigen, from 35.5 to 87.0 (P less than 0.001) with PHA (2 micrograms/ml), and from 107.2 to 181.9 (P = 0.007) with PHA (10 micrograms/ml). LT responses to C. trachomatis were compared in 52 pregnant women and 58 nonpregnant women; all the women had C. trachomatis isolated at the time of LT assay. Using either plasma supplement, the mean LTSI with C. trachomatis antigen was significantly higher in nonpregnant women than in pregnant women, regardless of trimester (P less than 0.001). Among 12 women who were serially tested and remained culture positive for C. trachomatis throughout pregnancy and the postpartum period, the mean autologous LTSI rose from 1.9 in the third trimester to 7.8 postpartum (P = 0.0004). These data are the first to show that the immune response to an ongoing bacterial infection is depressed during pregnancy and to definitively document the depressed LT responses during human pregnancy.

Antigens, Bacterial↗

[Evaluation of cell-mediated immunity in the course of primary malignant tumors of the ovary. Cutaneous reactivity. (author's transl)].

The present possibilities of clinically evaluating cell-mediated immune potential using skin reactivity to microbial protein antigens (streptokinase-streptodornase; tuberculin; formalin-killed parotitis virus) together with a contact sensitized such as dinitrochlorobenzene are examined. A single evaluation index for the set of cutireactions is proposed and, as a practical example, the delayed hypersensitivity data based on intradermal and patch tests in 41 cases of primary malignant tumor of the ovary are reported. There is a definite correlation between immune impairment and cancer spread, with a survival prognosis value even in initial cases.

Dinitrochlorobenzene↗

Varidase: the science behind the medicament.

Varidase is used throughout the world for the topical treatment of purulent and suppurating wounds. Its efficacy is centred on two enzymes, streptokinase and streptodornase. However, these represent only a small proportion of the bulked solid. This article gives an overview of the preparation and mode of action of Varidase, as well as showing some of the research and development that has gone into improving the assessment of its quality and composition.

Humans↗

Skin reactivity and phagocytic function of neutrophil leucocytes in Crohn's disease and ulcerative colitis.

A decreased capacity of the leucocytes to phagocyte yeast particles was found in patients with Crohn's disease of the ileum. Patients with lesions only in the colon showed normal phagocytic activity as did patients with ulcerative colitis (U.C.). Intradermal injection of killed Strep. pyogenes produced an increased erythrmatous reaction after 48 hours in patients with U.C., and the reaction was pustular in half of them. None of the controls or the patients with Crohn's disease of the ileum had such a reaction. An increased erythematous reaction to streptokinase-streptodornase was also more common in patients with U.C. than in those with Crohn's disease and a control group. No such difference was seen to other bacterial antigens. The reactivity to kallikrein, prostaglandin E1, histamine or bradykinin did not differ from that of normal subjects.

Adolescent↗

Topical treatment of pressure ulcers. A randomized comparative trial of Varidase and zinc oxide.

In a single-blind, randomized trial, the efficacy of topical streptokinase-streptodornase (Varidase) solution was compared with that of zinc oxide on necrotic pressure ulcers in 28 patients. The effectiveness was determined by measuring the necrosis removal within 8 weeks. This occurred in 6 patients (43%) treated with Varidase and in 7 (50%) treated with zinc oxide. The statistical tests applied showed no significant difference between the two treatments despite the use of a high power (1-beta = 0.95). The data suggest that the two regimens are about equally effective in the treatment of necrotic tissue.

Aged↗

Cell-mediated immunity in the rheumatoid diseases. I. Skin testing and mitogenic responses in sero-negative arthritides.

Cellular immunity has been investigated in patients with various kinds of sero-negative arthritis. The incidence of cutaneous response to recall antigen streptokinase-streptodornase (SK-SD), and the ability to mount a primary cutaneous response to dinitrochlorobenzene (DNCB) have been examined in patients with ankylosing spondylitis and psoriatic arthritis. The results were not significantly different from normal. In vitro lymphocyte transformation in the presence of phytohaemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM) has been measured using peripheral blood lymphocytes from patients with ankylosing spondylitis, psoriatic arthritis and Reiter's disease. In comparison with a control group, significantly reduced responses were found to a low dose of PHA in the ankylosing spondylitis and Reiter disease patients. Significant increase in response occurred to a high dose of PHA, in patients with psoriatic arthritis and Reiter's disease, and to PWM in Reiter's disease patients. The in vitro results in the ankylosing spondylitis, psoriatic arthritis and Reiter's disease patients suggest some abnormality in the T-cell population in sero-negative arthritis.

Adult↗

Stimulation of lymphocytes by antigen in microplate cultures; absence of an effect of transfer factor in vitro.

The best conditions for the optimum stimulation of human leucocytes by antigens, including protein purified derivative (PPD), streptokinase-streptodornase varidase (SKSD) and tetanus toxoid have been studied in microplate cultures. The leucocytes of each donor have their own characteristic response to antigen which depends on the culturing conditions such as antigen concentrations, cell concentrations and the time of measuring the rate of DNA synthesis. Thus, no conditions provide a universal optimum for antigen stimulation in vitro. Leucocyte dialysates, i.e. potential transfer factors, have been prepared from donors, who between them are strongly positive to the antigens PPD, SKSD, tetanus toxoid, diphtheria toxoid and Keyhole limpet haemocyanin. In contrast to some previous reports these leucocyte dialysates had no effect on the thymidine incorporation by leucocytes grown in the presence of these antigens. It is suggested that the selection of optimal conditions for the response to antigen may have obscured the effect of any non-specific enhancement of reactivity b lyeucocyte dialysates.

Adult↗

Diagnosis and prognosis in colon cancer based on a profile in immune reactivity.

An immunologic profile consisting of measurements of circulating carcinoembryonic antigen (CEA), tumor antigen-induced inhibition of monomuclear cell migration (IMM) and skin reactivity to purified protein derivative, streptokinase-streptodornase, and mumps was assessed as a diagnostic and prognostic tool in 16 patients with colon cancer. Preoperatively, 10 of 14 patients tested had elevated CEA, 12 of 12 showed tumor antigen-induced IMM, and 10 of 11 failed to react to 2 or more recall antigens. Potential surgical cure (7 patients) was accompanied by normal CEA in 4, absent tumor antigen-induced IMM in all 7, and increased skin-test reactivity in 6. Disseminated cancer (9 patients) was associated with elevated CEA in all 9, with absent IMM in all 7 and with suppressed skin-test reactivity in 6 of 9.

Adult↗

Immunodeficiency in patients with non-Hodgkin lymphomas.

Seventy-one previously untreated patients with non-Hodgkin lymphomas were studied with several readilyvailable tests of immune function: number of peripheral blood lymphocytes, serum immunoglobulins, and delayed hypersensitivity to six recall antigens. The results were correlated to histology (Rappaport classification), stage (Ann Arbor classification), the presence of symptoms, and survival. As a group, 38 patients with diffuse lymphomas exhibited marked impairment in reactivity to five of six antigens (p less than 0.03 to p less than 0.001). In addition, lymphopenia and reduced levels of serum IgA were found in association with diffuse histiocytic lymphoma. Among patients with diffuse lymphoma, lymphocyte number and skin test reactivity tended to be greater in those with localized disease or without constitutional symptoms, and survival was superior for patients free of symptoms (p less than 0.01). As a group, 33 patients with nodular lymphoma had normal numbers of lymphocytes, lower levels of serum IgG and IgA, and significant impairment of reactivity to two antigens (streptokinase-streptodornase and mumps; p less than 0.01); reactivity to three other antigens (Candida albicans, coccidiodin, and tuberculin) was normal. Survival for patients with nodular lymphoma was superior (p less than 0.01) compared to those with diffuse lymphomas. In summary, severe immunodeficiency was found in patients with diffuse lymphoma (particularly diffuse histiocytic lymphoma), and definite but much less severe immunodeficiency was characteristic of patients with nodular lymphoma.

Adult↗

Effect of human monocytes and macrophages on Trypanosoma cruzi.

Studies were undertaken to determine whether Trypanosoma cruzi can invade and multiply within human monocytes and macrophages cultured in vitro and, if so, whether macrophages can be activated to inhibit the multiplication. A reticulotropic strain of T. cruzi was capable of infecting human monocytes and monocyte-derived macrophages. Intracellular multiplication was observed in both cell types when they were examined microscopically. An increase in the number of trypanosomes occurred in the supernatants as well, providing additional evidence of intracellular multiplication and cell disruption by the parasite. Activation of monocyte-derived macrophages was accomplished by incubating the monolayers in the presence of lymphocytes and streptokinase-streptodornase. These activated macrophages inhibited intracellular multiplication of T. cruzi and the number of T. cruzi in the supernatants of these monolayers was markedly decreased as well.

Adult↗