Modulation of cell-mediated alloimmunity by BCG. II. Induction of specific, nonadherent, non-T-killer cells by BCG and alloantigen.

The cellular basis of the augmentation of cell-mediated immunity (CMI) by systemically administered BCG was investigated in C57BL/6 mice. BCG pretreatment potentiated the CMI to alloantigens as measured by a 48-hour microcytotoxicity assay (MCA) against P815Y tumor cells. The effector cell was not a T-cell, as demonstrated by its lack of sensitivity to antithymocyte serum and complement and its failure to kill in a short-term 51Cr release assay. The effector cell also was not a classical macrophage, because it was not depleted by treatment with silica. CMI as measured by the MCA was consistently depleted by magnetic separation after incubation with carbonyl iron. The active cell(s) showed variable adherence properties. The augmentation of alloimmunity was alloantigen-specific. The specificity appeared to be due to the interaction between effector cells without inherent specificity and sensitized T-cells.

Modulation of cell-mediated alloimmunity by BCG. I. Antagonism and potentiation of immunosuppression caused by cytarabine.

The ability of iv administered BCG to antagonize immunosuppression caused by injection of the antimetabolite cytarabine (beta-cytosine arabinoside; ara-C) was investigated in C57BL/6 mice. Treatment with BCG 10 days before alloimmunization with killed L1210 tumor cells decreased spleen T-cell-mediated cytolysis against allogeneic P815Y tumor cells, as measured by a short-term 51Cr release assay, and potentiated immunosuppression due to ara-C. In contrast, spleen cell-mediated immunity (CMI) that was assayed by a 48-hour microcytotoxicity assay (MCA) was augmented by systemic BCG administered before alloimmunization. Pretreatment with BCG resulted in a complete and long-lasting protection against the immunosuppressive effects of ara-C on this CMI as measured by the MCA. Treatment with BCG after cytoreductive therapy resulted in a significant, although transient, reversal of immunosuppression. Depending on the type of response and thus the type of effector cell measured, BCG acts as a moderate immunosuppressive agent or a strong immunopotentiator of CMI.

Detection of class-specific antibodies of pemphigus vulgaris using a solid phase radioimmunoassay.

A sensitive solid phase radioimmunoassay was developed to detect class-specific pemphigus antibodies. COLO-16, a cell line derived from a human squamous cell carcinoma of the skin, was used as the substrate, since this cell line has been shown by indirect immunofluorescence to bind IgG antibodies in sera from patients with pemphigus vulgaris, but not from those with bullous pemphigoid. IgG antibodies to COLO-16 were significantly greater in sera from 17/24 patients (p less than 0.05) with pemphigus vulgaris than in sera obtained from a control population consisting of normal volunteers and patients with other dermatologic diseases. The relative binding of the positive sera correlated well (p less than 0.05, by Spearman's rank correlation test), with the endpoint determined by indirect immunofluorescence against monkey esophagus. This technique is 10-100 times more sensitive than immunofluorescence for the detection of pemphigus antibodies and will be useful in studying the cellular mechanisms controlling the synthesis of pemphigus antibody in vitro.

Partial purification and characterization of pemphigus-like antigens in urine.

The inhibition of indirect immunofluorescence was used to detect the presence of pemphigus-like antigens in urinary proteins. Antigenic material was present in 12 of 40 urine samples obtained from 16 patients with bullous pemphigoid, cicatricial pemphigoid, or pemphigus vulgaris. Antigen could not be detected in the urine of patients with carcinoma of the bladder. Partial purification was accomplished by ion-exchange chromatography and column gel filtration. Antigenic activity was detectable in all fractions eluted from Sephadex G-200 in the broad range of molecular weight of 90,000-180,000 daltons. Pemphigus-like antigens were glycoproteins with an apparent molecular weight of 75,000 daltons by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The antigens were immunogenic in mice, but were not detectable by double diffusion in agarose gels.

The 'no-touch' method of intermittent urinary catheter insertion: can it reduce the risk of bacteria entering the bladder?

OBJECTIVE: This in vitro model was designed to determine whether using a no-touch method for catheter preparation and insertion would affect the degree of contamination transmitted to intermittent urinary catheters. SETTING: Northview Laboratories, Northbrook, IL 60062, USA. METHODS: This was a parallel experimental study conducted in vitro at an independent testing laboratory under Good Manufacturing Practices. Six different models of intermittent urinary catheter were tested in triplicate, in accordance with the manufacturer's Instructions For Use (IFU). Gloved hands were contaminated with a known amount of Staphylococcus aureus or Escherichia coli, preparation for insertion was mimicked according to the manufacturer's IFU. The number of bacteria transferred to the catheter was then quantified using a validated technique. The null hypothesis tested was that the bacterial contamination resulting from handling would be the same for all of the catheters. The primary outcome measure was the amount of bacteria, expressed as colony forming units (CFU), recovered from the catheters. The catheter wrappers were sampled to confirm that active transfer of microorganisms had taken place and nonhandled samples served as the negative controls. RESULTS: For both test organisms, the number of bacteria recovered from the catheters was significantly lower (P < 0.05) for the catheters featuring the no-touch design (approximately 5 CFU/plate versus 2.1 x 10(2) to 4.4 x 10(2) for the traditional hydrophilic catheters). The bacterial recovery from those catheters with the no-touch design often fell below the nominal detection limit. CONCLUSION: The results of this study allow the null hypothesis to be rejected; it can be concluded that the no-touch method provides a significant benefit in reducing the potential for external contamination of an intermittent urinary catheter. This result supports the recent recommendations for aseptic intermittent catheterization promoted within the guidelines issued by the European Association of Urologists (EAU).

Effect of pH on the production of irritation in a chamber irritation test.

This study was designed to investigate the relationship between alteration of skin surface pH and the development of skin irritation. Application of a phosphate/borate/acetate buffer adjusted to pH 4.0-10.5 in a standard chamber irritation test did not result in significant clinical irritation. This was true in spite of maintaining a markedly altered skin surface pH over an extended period of time. There was also no correlation between irritation and the pH of applied surfactant solutions. These results demonstrate that a prolonged disturbance of the "acid mantle" is not sufficient to produce clinical irritation.

Clinical correlations of antibodies in pemphigus studied by radioimmunoassay.

Pemphigus antibodies of class G immunoglobulins (IgG) were studied with the use of a solid-phase radioimmunoassay (RIA) that utilizes COLO-16, a squamous cell tumor line, as the substrate. Sera from patients with pemphigus vulgaris and pemphigus vegetans showed greater average binding than did sera from patients with pemphigus foliaceus and pemphigus erythematosus. The patients were subgrouped according to their clinical presentations and were divided into patients with skin lesions only, those with mucous membrane lesions only, and those with both skin and mucous membrane lesions. No significant differences were observed between these groups. There was good correlation between IgG binding in the RIA and both the disease activity and the response to therapy. Blood group antigen absorbable "false positive" anti-intercellular substance antibodies were not detected in the RIA.

Detection of anti-intercellular cement substance and anti-basement membrane zone antibodies by radioimmunoassay using SCaBER tumour cell line as substrate.

IgG autoantibodies present in the serum of pemphigus vulgaris and bullous pemphigoid patients were detected by a solid phase radioimmunoassay, using a squamous cell tumour line, SCaBER, as substrate. This preliminary study shows that the SCaBER cell line displays both pemphigus and bullous pemphigoid antigens. This source of antigens should allow the development of a sensitive assay to measure anti-ICS and anti-BMZ antibodies. Such an assay may have clinical applications and may provide an important tool for studying the mechanisms of autoantibody production in pemphigus and pemphigoid.

The use of transepidermal water loss to measure and predict the irritation response to surfactants.

Synopsis The ability of baseline transepidermal water loss (TEWL) measurements to predict the response of healthy subjects to a standard soap chamber irritation insult was evaluated. Chambers containing surfactant solutions were applied to the forearms of normal volunteers for five consecutive days. TEWL was measured using a Servo Med Evaporimeter. TEWL measurements were made prior to the first application of the chambers and following the removal of the second set of chambers (the afternoon of Day 1). The correlation between the TEWL values obtained on Day 1 and the subjective evaluation of irritation resulting from the application of two different toilet soaps was statistically significant. This result indicates that damage to the skin is reflected by an increase in TEWL. Interestingly, there was also a statistically significant correlation between TEWL values obtained on the untreated skin at Time 0 and the five-day average irritation score for these two cleansing products. The results of this study indicate that a high baseline TEWL value may be predictive of a high susceptibility to soap irritation as measured by the chamber test. Le role des pertes en eau transepidermique pour mesurer et prevoir l'irritation due aux tensio-actifs.

Objective measurement of surfactant irritation by fiber optic spectroscopy.

Erythema caused by occlusion of 3 concentrations of surfactant was quantified in vivo using a modified spectrophotometer. Standard chamber irritation test sites were evaluated objectively using the spectrophotometer and subjectively by trained observers. There was a good relationship between the instrumental and visual evaluations. The spectrum was analyzed and a region was chosen to optimize the sensitivity of the method.

Prototypes of facial attributes developed through image averaging techniques.

Image capture and quantification has proven useful in a variety of scientific applications, for example, biology, medicine, geology, meteorology and forensics. The objective of this research was to utilize this technology to quantify clinical- and consumer-perceivable changes in facial attributes. A panel of expert assessors was trained, and, in a large consumer study, consumer facial attributes were identified and grading scales for each attribute were established. These experts then rated over 240 subjects on a total of 19 different facial attributes. Based on methodology developed by Perrett et al., facial averages or prototypes were computed from panelists rated high or low for each attribute. Prototypes were developed in a 3 step process: 1) selection of 224 predefined feature points; 2) calculation of average face shape; and 3) 'morphing' individual faces into that shape and blending the images together. Naive assessors could readily appreciate the differences in facial appearance of the prototypes. In addition, expert graders were able to identify the general class of attribute affected. This method provides a powerful tool for assessing the effects of skin care technologies.