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M J Cruz

Publications and source records attributed to M J Cruz.

4 recordsLinked to original sources

Blood sample processing effect on eosinophil cationic protein concentration.

BACKGROUND: Asthma is considered to be an inflammatory disease. The most important cell involved in the inflammation is the eosinophil. These cells and their mediators, such as eosinophil cationic protein (ECP), are potential markers of the inflammation's severity. Eosinophil cationic protein may be used for monitoring antiasthma treatment. It is well known that sample processing conditions can affect the ECP blood levels. OBJECTIVE: The aim of this work is to study the effect of temperature, time, and anticoagulants on ECP levels. METHODS: We studied five asthmatic patients and five healthy controls. We obtained three different blood samples from each subject, one with heparin, one with EDTA, and one without anticoagulant. To evaluate the effect of temperature, serum samples were clotted for an hour, one at 0 degree C, one at room temperature, and the other at 37 degrees C. Plasma (heparin and EDTA) samples were treated as follows: one was immediately centrifuged, and two were stored for an hour, one at 0 degree C, and the other at room temperature. Eosinophil cationic protein levels were measured by fluoroimmunoassay (CAP-System ECP FEIA Pharmacia). RESULTS: A higher temperature during blood clotting resulted in a higher ECP concentration. There were no differences between ECP determination in serum samples and plasma samples with heparin, under the same conditions of time and temperature; so clotting may not be necessary for ECP release in vitro. Eosinophil cationic protein was not released in plasma samples with EDTA, neither at 0 degree C nor room temperature. CONCLUSIONS: More studies must be done to clarify the mechanism of the ECP release in vitro.

Adult

Characterization of specific immunoglobulin G (IgG) and its subclasses (IgG1 and IgG2) against the 23-valent pneumococcal vaccine in a healthy adult population: proposal for response criteria.

The aim of the study was to standardize an enzyme-linked immunosorbent assay (ELISA) method for the quantification of immunoglobulin G (IgG) and its subclasses (IgG1 and IgG2) against the 23-valent pneumococcal vaccine and to establish the criteria for a normal response to the vaccine. Forty healthy individuals (20 women and 20 men; mean age, 29 years) were studied. All were vaccinated with the 23-valent pneumococcal vaccine; blood samples were drawn just prior to and 3 weeks after immunization. Quantification of specific IgG and its subclasses was performed by an ELISA with the vaccine as the antigen. The linearity of the ELISA method was demonstrated by the similar slopes of the linear regression lines generated from the titration of sera with different antibody concentrations. The specificity of the antibodies against the vaccine was demonstrated by (i) an absorption test with pneumococcal vaccine, (ii) a cross-reactivity experiment with Haemophilus influenzae type b polysaccharide, and (iii) affinity chromatography with protein A-Sepharose. Response to the vaccine was defined by using the lower level of the 90% probability interval (one-tailed) for postimmunization-specific IgG, IgG1, and IgG2. By using this cutoff, responders were considered to be those with an absolute increase in antibody titers higher than 395 arbitrary units/ml for IgG, 0.350 A450 units for IgG1, and 0.314 A450 units for IgG2. Overall, 20 (50%) subjects had IgG, IgG1, and IgG2 responses, 9 (22.5%) had IgG and IgG2 responses, 4 (10%) had IgG1 responses, 3 (7.5%) had IgG and IgG1 responses, and 4 (10%) were nonresponders. Ninety percent of our population responded to the 23-valent pneumococcal vaccine. Up to 10% of healthy individuals may respond to an IgG subclass without significant increases in total IgG titers. The ELISA method that is described may be useful for evaluating the specific antibody response against polysaccharides.

Adult

IgG subclass deficiencies associated with bronchiectasis.

Only a small number of patients with IgG subclass deficiencies (IgGSD) have been observed to have bronchiectasis. Moreover, in the series of patients with bronchiectasis, IgGSD have not been found at any frequency, and the etiology of bronchiectasis remains unclear in 29 to 49% of cases. Serum concentrations of total IgG, IgA, and IgG subclasses as well as pulmonary function were measured in 65 patients (aged: 10 to 74 yr) with bronchiectasis of unknown etiology. An ELISA test was performed to quantify subclasses 1 through 4 using subclass-specific antihuman monoclonal antibodies. IgG subclass estimation in a healthy population with age-stratified normal ranges was derived from 100 adults, 37 children aged between 10 and 12 yr, and 27 adolescents aged between 13 and 16 yr. Serum concentrations of specific IgG antibodies to Haemophilus influenzae type b capsular polysaccharide (Hib-PRP) were also assayed by an ELISA test in 19 of the patients (10 with IgGSD and nine with non-IgGSD) and in 58 healthy individuals before and 3 wk after immunization with Hib-PRP conjugated to meningococcal outer membrane protein complex (OMPC). Thirty-one patients (48%) had low serum concentrations of one or more IgG subclasses (19 IgG2 deficiencies, 2 IgG3 deficiencies, 3 IgG4 deficiencies, and 7 combined subclass deficiencies). All patients showed increased levels of total IgG, IgG1, and IgA, but this rise was significantly higher in patients without IgGSD. Patients with IgGSD showed impaired antibody response to Hib-PRP compared with patients with non-IgGSD and the control group. IgGSD, particularly IgG2 deficiency, are not an unusual cause of bronchiectasis. Therefore, serum levels of IgG subclasses must be assayed whenever other causes of bronchiectasis have been ruled out.

Adolescent