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S Gallardo

Publications and source records attributed to S Gallardo.

35 records · Page 2Linked to original sources
PubMed1997

Study of apoptosis in human lymphocytes by toxic substances implicated in toxic oil syndrome.

Toxic Oil Syndrome is a multisystemic disease that occurred in epidemic proportions in Spain in 1981 caused by the ingestion of rapeseed oil denatured with aniline. Several data implicate T cells in the pathogenesis of the disease. We evaluated the mechanisms of cytotoxicity in human lymphocytes of TOS-related products: aniline, 3-(N-phenylamino)-1,2-propanediol and its mono- and di-oleyl esters and eosinophilia myalgia-related product such as 3-(phenylamino)-L-alanine, which is chemically similar to 3-(N-phenylamino)-1,2-propanediol, and has been found in manufactured L-tryptophan. Our results show that only di-oleyl ester of 3-(N-phenylamino)-1,2-propanediol induces apoptosis in human lymphocytes, in a concentration and time-dependent way, confirmed by morphology, expression of phosphatidylserine in membrane and analysis of DNA degradation.

Alanine↗
PubMed1997

Eosinophils transcribe and translate messenger RNA for inducible nitric oxide synthase.

Because of the involvement of nitric oxide (NO) in inflammatory states such as parasitic and hypersensitivity disorders and the fact that eosinophils are one of the cell types implicated, we asked whether eosinophils were able to express mRNA specific to inducible NO synthase (iNOS) and iNOS protein and to secrete nitric oxide. iNOS protein was detected on eosinophil preparations by immunocytochemistry using iNOS mAb. Expression of iNOS protein was also detected by immunoblotting in human purified eosinophils and an eosinophilic leukemia cell line, Eol-3. Nitrite production was detected in the supernatant of human eosinophils and Eol-3 cells cultured for 24 h, and was completely inhibited in the presence of the NOS inhibitor N-methylester-L-arginine. iNOS cDNA was obtained by reverse transcription-PCR. After subcloning, sequencing of the 259-bp fragment from three different human eosinophils cDNAs revealed 97% identity with macrophage/monocyte iNOS. Our studies describe for the first time the presence of iNOS on eosinophil and a putative new role for this cell in inflammatory states such as asthma and parasitic disease.

Animals↗
PubMed1997

Autoimmune hepatitis type 2 and hepatitis C virus infection: study of HLA antigens.

BACKGROUND/AIMS: Markers for hepatitis C virus are often detectable in patients suffering chronic hepatitis with liver-kidney microsomal type 1 antibodies. Several authors have suggested that two subsets of those patients can be defined: a) hepatitis C virus negative and b) hepatitis C virus positive. The aim of this work was to further analyze the possible genetic association, HLA class I and II, in these two groups of patients. METHODS: HLA was analyzed in 49 patients. Class I was studied using a standard lymphocytotoxicity test and in class II a reverse hybridization-based test for DRB1 typing and PCR-SSO for DQB1 typing were used. Sixty healthy Spanish subjects and 39 chronic hepatitis C subjects without anti-LKM1 antibodies were used as control groups for the "a" and "b" subsets, respectively. RESULTS: No significant association was found with class I specificities in either group. DQB1 typing showed a very significant increase of DQ2 in the "a" group (93.3% vs. 48%; RR = 15; Pc = 0.0025), and DRB1 typing from the "b" group revealed a high association with DR7 (82.3% vs. 43.6%; RR = 6; Pc = 0.0086). CONCLUSIONS: Our studies revealed a strong association with DQ2 for the "a" group and for the first time an extremely high association with DR7 antigen for the "b" subset. Hence it is possible to establish a different genetic profile in these two patient groups.

Adolescent↗
PubMed1996

Allergy to olive pollen: T-cell response from olive allergic patients is restricted by DR7-DQ2 antigens.

BACKGROUND: We have recently described the association between the IgE antibody response to Ole e I (the major antigen from olive tree pollen) and the DR7-DQ2 haplotype in a Spanish population. OBJECTIVE AND METHODS: Due to the linkage disequilibrium between DR7 and DQ2, and thus the difficult distinction between the role of these two antigens in the T-cell activation response, we decided to solve this question by two approaches: 1. The study of another ethnic group, individuals of Arabic origin, with a presumably distinct disequilibrium linkage between DR and DQ antigens. Genomic DNA typing was performed in 46 subjects (allergic and non-allergic) by Restriction Fragment Length Polymorphism (RFLP) and results showed that patients with specific IgE antibodies alpha-Ole e I, were DR7 and/or DQ2. These data show a similar restriction pattern to those previously described for Spanish patients. The phenotypic frequency of DR7 antigen is significantly greater than in the non-allergic population, with a corrected P (Pc) value of 0.03. 2. The analysis of the genetic requirements of Ole e I response, using T-cell lines specific for this antigen. This was first carried out by blocking the proliferative response of these T-cell lines with specific anti-human HLA class II antibodies and then testing the genetic restriction of this response using a panel of histocompatible and histoincompatible Antigen Presenting Cells (APCs). Both experiments corroborate the hypothesis that DR7 and DQ2 are implicated in the recognition of Ole e I.

Adolescent↗
PubMed1994

Immunological basis of toxic oil syndrome (TOS).

The toxic oil syndrome (TOS), a multisystemic disease, that occurred in Spain in 1981, was caused by the ingestion of rapeseed oil denatured with 2% aniline. Due to the clinical course of the disease, immunopathological mechanisms have been suspected but a direct connection was never demonstrated. To analyse this possibility, we determined several immunological parameters in the sera of patients with TOS and without the disease, using a case-control design: total immunoglobulins, IgG and IgE antibodies against different toxic agents (oleylanilide, aniline, linoleyl-anilide, and 3-phenylaminopropane-1-2-diol), autoantibodies, cytokines (IL-4, IL-6, TNF, GM-CSF) and soluble receptors (sCD23 and sIL-2R). We detected high levels of sIL-2R in TOS patients compared to controls (P < 0.0001). A higher levels of sCD23 and IgE were also found. In addition, the response to oleyl-anilide of peripheral blood lymphocytes from TOS patients was studied and a significant proliferative response in 30% of TOS patients versus 5% controls was observed. Our data support the implication of the immune system in the acute phase of TOS, with a possible activation of T-cells and release of cytokines, that could explain some of the clinical findings in this phase of the disease.

Anilides↗
PubMed1994

Ole e I: epitope mapping, cross-reactivity with other Oleaceae pollens and ultrastructural localization.

Ole e I is the major allergen derived from olive tree pollen (Olea europaea) and it is composed of two polypeptides with molecular weights (MWs) of 18 and 20 kD. A panel of six monoclonal antibodies (mAbs) has been prepared and used to map antigenic determinants on this molecule. Four epitope determinants have been identified on Ole e I. Using the purified mAbs produced against Ole e I, we have analyzed the common epitope determinants in olive (O. europaea) and different Oleaceae pollens: ash (Fraxinus excelsior); privet (Ligustrum vulgare); lilac (Syringa vulgaris), and forsythia (Forsythia suspensa). ELISA showed three reactivity groups depending on the recognition of monoclonal antibodies: (1) olive and ash; (2) olive, ash, privet and lilac; and (3) olive, ash, privet, lilac and forsythia. Immunoblotting studies on Oleaceae pollen extracts with these mAbs showed a very similar cross-reactivity pattern. The 18- and 20-kD MW proteins were present in each pollen, except in the case of forsythia. In this case the reactivity pattern was associated with 50- to 55-kD protein bands. This band was recognized by a pool of sera from olive-allergic patients. Finally, ultrastructural localization of Ole e I antigen was performed on the mature olive pollen grain. Ole e I was located in association with dilated endoplasmic reticulum cisternae. Pollen grain walls, nuclei and cytoplasmic organelles were totally devoid of the allergen.

Allergens↗
PubMed1994

Modulation of the Fc gamma RII and Fc gamma RIII induced by GM-CSF, IFN-gamma and IL-4 on murine eosinophils.

Murine eosinophils express the low-affinity Fc gamma receptors recognized by the monoclonal antibody (mAb) 2.4G2, which are involved in the activation of these cells. We have studied the membrane and RNA expression of the low-affinity Fc gamma receptors on murine eosinophils stimulated by granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon-gamma (IFN-gamma), and interleukin-4 (IL-4). Flow cytometric analysis (FACS) of eosinophils incubated with GM-CSF and IFN-gamma showed an up-regulation on the expression of these receptors with a maximal effect at 24 hr. IL-4 failed to induce any positive or negative effect in these cells. To assess the pattern of expression of the low-affinity Fc gamma receptors on eosinophils, Northern blot analyses were carried out using specific cDNA probes encoding for the Fc gamma RII and Fc gamma RIII. Murine eosinophils constitutively express transcripts for Fc gamma RII and RIII. Incubation with GM-CSF from 3 to 12 hr produced a potent induction of the two transcripts studied (Fc gamma RII and RIII). IFN-gamma did not induce any important up-regulation of the two transcripts from 3 to 12 hr of incubation. By contrast, IL-4 produced a marked inhibition of both transcripts at 24 hr of incubation. Expression of the gamma-chain transcript which is associated with Fc gamma RIII has been detected on freshly isolated eosinophils. This study demonstrates a different regulation pattern of these receptors depending on the cytokine or growth factor used to stimulate murine eosinophils.

Animals↗
PubMed1993

DR7 and DQ2 are positively associated with immunoglobulin-E response to the main antigen of olive pollen (Ole e I) in allergic patients.

We have studied the relationship between HLA class II haplotypes and alleles, and the IgE antibody response to a highly purified allergen, Ole e I, in allergic patients. Ole e I, is the major antigen from the pollen of olive tree that grows mainly in the Mediterranean. Genomic DNA typing was performed in 40 unrelated patients with seasonal allergic pollenosis who had specific IgE antibodies against Ole e I, detected by double-antibody radioimmunoassay. HLA-DRB and -DQB loci were analyzed by PCR-SSO and RFLP. Phenotypic frequencies were compared with those of 179 healthy unrelated individuals. Significant increases in the phenotypic frequencies of DR7 (pf = 67.5% vs 31.3% in the control population, pc = 0.0023) and DQ2 (pf = 90.0% vs 48.0%, pc = 0.0003) were found, indicating an association between DRB1*0701/2, DQB1*0201 alleles and the IgE antibody response to Ole e I. This is the first time that the HLA-DQ gene has been associated with a positive allergic response.

Allergens↗
PubMed1992

Eosinophil as antigen-presenting cell: activation of T cell clones and T cell hybridoma by eosinophils after antigen processing.

We have studied the role of murine eosinophils as antigen-presenting cells (APC). Eosinophils have several characteristics that support the hypothesis of its function as potential APC: they have phagocytic capacity, express adhesion molecules and major histocompatibility complex (MHC) class II antigens and can produce and release interleukin-1 (IL-1). We have obtained several T cell clones specific for Mesocestoides corti antigens and used T cell hybridoma specific for ovalbumin (OVA) to test this hypothesis. Granulocyte-macrophage colony-stimulating factor-activated pure eosinophils (99.9%), express class II antigens and are able to present M. corti antigens to specific T cell clones or OVA to T cell hybridoma 3DO 11.10, inducing the proliferation of T cell clones and IL-2 release by the T cell hybridoma. Proliferation of T cells clones is dependent on the number of eosinophils used as APC. We have compared the efficiency of the same number of macrophages and eosinophils as APC, and have found that macrophages are more efficient than eosinophils. Lysosomotropic agents, such as chloroquine and ammonium chloride, that inhibit antigen processing, impaired eosinophil presentation. This presentation is restricted by MHC class II and inhibited by anti-I-Ad monoclonal antibody. The present study provides clear evidence of APC function for eosinophils. Our investigation points to a new role for eosinophils in the immune response.

Ammonium Chloride↗
PubMed1992

[Utility of high-resonance ultrasonography in the diagnosis of acute appendicitis].

The clinical records and the ultrasound findings of 48 patients studied because of a presumptive diagnosis of acute appendicitis in a one-year period were reviewed. Ultrasound examination was performed using graded compression and high resolution probes when acute appendicitis was suspected and the clinical history or physical examination was unclear. The ultrasound findings were correlated with the clinical course or surgical and pathological findings. This procedure was useful in the diagnosis of acute appendicitis, with 84.6% sensibility and 95.5% specificity. Predictive value for positive results was 95.7% and 84% for negative results. These figures agree with results previously communicated in the literature. It is concluded that high resolution ultrasonography is useful in the differential diagnosis of atypical acute appendicitis.

Abdomen, Acute↗
PubMed1990

[Percutaneous drainage of abdominal abscesses].

Percutaneous drainage has been widely accepted as the preferred treatment for abdominal abscess. Indications have been clarified recently and the absence of a secure route is the only absolute contraindication. We performed this procedure in 65 patients with abdominal abscess at different locations: liver 27, subphrenic 33, lesser cavity 3 and perirenal 3. Overall success rate was 85%, with 89% for liver and 88% for the subphrenic location. Six patients died of multisystemic failure even though the abscess was properly drained. Four patients were operated on for persistent abscess. A pancreatic fistula was shown in 1 and a peritoneal hydatid cyst was the original lesion in the other. Pneumothorax occurred in 5 patients requiring drainage in 2. Two other patients developed hydro-pneumothorax and empyema, that was drained. The overall complication rate was 14%. Thus, percutaneous drainage is a simple and highly successful treatment for abdominal abscess. Results are influenced by the accuracy of diagnosis and a proper selection of patients.

Abdomen↗