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Biomedical subjects

P Aparicio

Publications and source records attributed to P Aparicio.

At least 37 records · Page 2Linked to original sources

Ribotyping of Staphylococcus aureus: an assessment using well-defined strains.

Ribotyping, with homologous or heterologous (Escherichia coli) r-RNA, of the propagating strains for phages of the international set for strains of Staphylococcus aureus of human origin was undertaken to determine the discrimination of this typing method. Ribotyping could distinguish between strains of different phage groups, but could not distinguish between seven phage group III strains of different phage type. Ribotying may be a useful adjunct to phage typing in S. aureus but is unlikely to replace it as the primary method of epidemiological typing.

Bacterial Typing Techniques↗

An additional set of phages to characterize epidemic methicillin-resistant Staphylococcus aureus strains from Spain (1989-92).

In recent years, methicillin-resistant Staphylococcus aureus (MRSA) isolates in Spain have increased dramatically; in 1986 there were only 1.2% MRSA amongst all nosocomial Staphylococcus aureus (SA) isolates, by 1989 this percentage had risen to 44% in some hospital causing a very serious epidemic situation in the country. We have characterized these isolates by direct, reverse and Fisk phage typing and we have also looked for an additional local set of phages to help us to differentiate these strains. We have been able to differentiate an epidemic strain from other MRSA strains which cause sporadic hospital outbreaks, and we have also distinguished between some variants of the epidemic strain.

Bacteriophage Typing↗

A novel functional cell surface dimer (kp43) serves as accessory molecule for the activation of a subset of human gamma delta T cells.

Kp43 is a cell surface molecule expressed by NK cells, gamma delta T lymphocytes, and a small subset of CD56+ alpha beta T cells. In the present study the distribution and function of Kp43 on gamma delta T cells is described. The expression of Kp43 on freshly isolated gamma delta T-cell populations derived from fetal and adult tissues was biased toward memory V gamma 2+/V delta 2+ T lymphocytes and appeared to be up-regulated after activation. Several gamma delta T-cell clones were derived from different sources. Adult V gamma 2+/V delta 2+ T-cell clones expressed the highest levels of Kp43, V delta 1 T-cell clones derived from periphery expressed moderate levels, whereas some thymus V delta 1 T cells and fetal gamma delta T-cell clones displayed low or undetectable levels of Kp43 molecule. Based on the effect of a specific mAb, this dimer appears to be fully functional as previously shown for NK cells. The stimulation of Kp43+ gamma delta T-cell clones with a combination of anti-CD3 and anti-Kp43 co-immobilized on a solid substratum enhanced both the expression of cell surface CD25 and the secretion of TNF-alpha. The restricted expression and functional activity of the Kp43 dimer suggests that it may play an important regulatory role in activation, selection, Ag recognition, and responsiveness of a subset of gamma delta T cells.

Antigen-Presenting Cells↗

ICAM-3, the third LFA-1 counterreceptor, is a co-stimulatory molecule for both resting and activated T lymphocytes.

Optimal activation of human T cells mediated by ligation of CD3/T cell receptor (TcR) complex requires co-stimulatory signals. These can be provided by the adhesive interaction between receptor molecules on T cells and their counter-receptors on antigen-presenting cells. Soluble ICAM-3, anti-ICAM-3 and anti-CD3 mAb were utilized to address the role of the ICAM-3/LFA-1 pathway in TcR/CD3-dependent or -independent T cell activation. Immunoaffinity-purified ICAM-3 co-immobilized with suboptimal concentrations of anti-CD3 monoclonal antibody (mAb) stimulated T lymphocytes as monitored by the expression of the lymphocyte activation antigens CD25 and CD69. The mechanism underlaying this activation appear to involve the interaction of ICAM-3 with a beta 2 integrin, likely to be LFA-1, since mAb to the CD18 chain completely inhibited T cell activation. Similar experiments demonstrated that anti-ICAM-3 mAb were able to co-stimulate both resting (cord blood) and activated (T cell clones) T lymphocytes. On the contrary, anti-ICAM-1 mAb were only co-stimulatory for CD25 expression on activated but not on resting T cells. In addition, we have found that some gamma delta T cell clones bearing the V delta 1 segment were activated by direct mAb engagement of ICAM-3 in the absence of TcR/CD3 occupancy. Furthermore, immobilized anti-ICAM-3 mAb also induced development of dendritic processes. In conclusion, our data suggest that ICAM-3 on the surface of both T cells and antigen-presenting cells plays an essential role in the initiation of the immune response.

Adult↗

The CD26 antigen is coupled to protein tyrosine phosphorylation and implicated in CD16-mediated lysis in natural killer cells.

The levels of CD26 expression, their capacity to induce protein tyrosine phosphorylation and their functional implication in natural killer (NK) cytolysis have been studied. It was found that only a small fraction (12-15%) of peripheral NK cells expresses CD26 compared with the high expression (99%) found in NK clones. The protein tyrosine phosphorylation mediated by means of CD26 activation was studied in NK cells treated with the anti-CD26 MoAb 134-2C2, and two new proteins of 50 and 21 kDa appeared phosphorylated in tyrosine residues. To study the influence of CD26 antigen in NK lysis, we analysed the lytic capacity of NK cells stimulated with different anti-CD26 MoAbs or after separation into CD26+ and CD26- subsets and using K562 as target cells. Under these conditions, no differences were found in the chromium release by the target cells. Redirected lysis through CD16 was also measured by arming the effector cells (CD26+ and CD26-) with anti-CD16 antibody and using K562 as target cells. It was found that CD26- cells showed significantly less CD16-dependent lysis than CD26+ cells. These results indicate that CD26 is related to the CD16-dependent lysis but not to NK cytolysis which may be caused by mediation of protein tyrosine phosphorylation.

Antibodies, Monoclonal↗

An epidemic methicillin-resistant strain of Staphylococcus aureus in Spain.

During 1990, a strain of methicillin-resistant Staphylococcus aureus became epidemic in Spain and spread in a manner analogous to that of EMRSA-1 in England. Isolates of this strain produced little protein A and were resistant to a number of antibiotics including ciprofloxacin. Beta-lactamase production was encoded by a c. 39 kb plasmid, which also conferred resistance to mercury, cadmium, ethidium bromide and propamidine isethionate. Investigation showed that two variants, separable by supplementary and Fisk phage typing, were circulating. The B variant appeared to spread more readily than the A variant. The opportunity was taken to compare the discriminatory power of traditional typing methods with molecular techniques. The discriminatory power of the molecular techniques used only reached the same level as the traditional methods when double enzyme digestion of total cellular DNA by EcoR I and Cla I was performed.

Bacterial Typing Techniques↗

Functionally distinct subsets of human gamma/delta T cells.

To determine if effector subsets exist among human gamma/delta T cells, we examined the cytokine production and cytotoxic activity of gamma/delta T cell clones with different accessory molecule phenotypes, V delta and V gamma gene expression, and J gamma rearrangements. T cell clones bearing gamma/delta T cell receptor produce an array of cytokines like alpha/beta T cell clones. Individual gamma/delta T cell clones produced a characteristic array of cytokines without correlation with V delta or V gamma gene expression. However, when phenotypic subsets were considered, CD4+ gamma/delta clones produced significantly higher levels of interleukin 2 and granulocyte-monocyte colony-stimulating factor compared with CD4-CD8- and CD8+ gamma/delta clones. Similarly, when cytotoxic potential was assessed, CD4+ gamma/delta clones exhibited minimal activity when compared with CD4-CD8- and CD8+ adult peripheral blood gamma/delta clones. We conclude that functionally distinct gamma/delta T cell subsets exist and suggest that these subsets may correlate with expression of the CD4 accessory molecule.

Amino Acid Sequence↗

Low IgG2 and polysaccharide response in a T cell receptor expression defect.

B lymphocytes require appropriate T lymphocyte cooperation to synthesize immunoglobulins (Ig). Such interaction presumably takes place after engagement of the T cell receptor (TcR) by antigen. The present work addresses B lymphocyte function (and phenotype) in a novel type of immunodeficiency which is characterized by a TcR expression defect. In contrast to expectations, the two affected siblings that were studied displayed normal in vivo antibody responses to both endogenous and exogenous protein antigens. However, they showed impaired responses to certain polysaccharide antigens together with a selective IgG2 deficiency. These results suggest that some polysaccharide responses may be more T cell dependent than previously suspected, and support the notion that T cell dysfunctions (of this or other kind), rather than Ig gene deletions, may be the molecular basis of certain IgG2 deficiencies. To rule out a concomitant gross B cell dysfunction in these individuals, B lymphocyte phenotype and function were assayed in vitro, and found to be normal. A T cell line derived from one of the siblings displayed an abnormal TcR on the cell surface, but it showed several normal TcR-mediated functions. This suggests that the low number of peripheral T lymphocytes that have been found to express low TcR levels in these immunodeficiencies may be operational, and supplying sufficient "help" for the observed normal antibody responses to all tested protein, but not polysaccharide, antigens.

B-Lymphocytes↗

An experimental model of chronic osteomyelitis caused by Escherichia coli treated with cefotaxime.

An experimental model in Wistar rats, of osteomyelitis caused by Escherichia coli, was used to evaluate the efficacy of cefotaxime in two treatment regimens of different durations. Four groups of rats were set up: a group of rats receiving short-term treatment (14 days) with subcutaneous cefotaxime (100 mg bd), killed after 56 days; a control group receiving no treatment, killed after 56 days; a group of rats undergoing long-term treatment (28 days) with subcutaneous cefotaxime as above, killed after 70 days and a control group of rats receiving no treatment, killed after 70 days. Analysis of histopathological and microbiological findings revealed significantly better results in the long-term treatment group. No side-effects were observed during treatment or afterwards.

Animals↗

Cefotaxime in the treatment of chronic osteomyelitis caused by gram-negative bacilli.

Fifty patients with chronic osteomyelitis due to aerobic or facultative Gram-negative bacilli, alone or in mixed infections with Gram-positive cocci, were treated with cefotaxime. The diagnosis of osteomyelitis was made on the basis of clinical, roentgenographical, isotopic, microbiological and histopathological evidence of infection. Only those patients with infection sensitive to cefotaxime were treated. Most patients received 2 g cefotaxime qid iv for 30-60 days (mean 40 days). Some patients received metronidazole in addition. The following results were obtained at the six month follow-up: 40 patients cured, six improved, no failures and four relapses. The surgical technique had an impact on these results. No clinically important side-effects were encountered, nor were any cefotaxime resistant strains found.

Adolescent↗

[Influence of bacterial combinations on the post-antibiotic effect].

A study of the post-antibiotic effect (PAE) and letality curves of different concentrations of clavulanic acid, amoxycillin, amoxycillin + clavulanic acid, netilmycin and ofloxacin on mixed cultures of S. aureus and E. coli. The post-antibiotic effect was measured after one hour of exposure of bacteria to antimicrobial and the elimination of the latter by the dilution method. Amoxycillin did not induce PAE on the association. PAE only appeared at high concentrations on S. aureus when evaluated alone. Clavulanic acid induced a greater PAE than amoxycillin. The association clavulanic acid + amoxycillin was synergistic and induced a greater PAE than its components. Netilmycin and ofloxacin were the drugs that induced the greatest PAE on the bacterial association and on its strains independently evaluated.

Amoxicillin↗

Isolation and characterization of (gamma, delta) CD4+ T cell clones derived from human fetal liver cells.

Lymphocytes isolated from human fetal liver and expanded in vitro in IL-2-containing media reveal the existence of CD4+ gamma, delta T cells. These cells display differential features of double-negative and CD8+ gamma, delta T cells as well as of CD4+ alpha, beta T cells. Thus, they failed to lyse targets in lectin-mediated killing assays and to perform classical helper functions. These results add new information necessary for a better understanding of the physiological role of the gamma, delta T cells.

Antigens, Differentiation, T-Lymphocyte↗

CD8 is involved in both class I- and class II-induced proliferation of a cytolytic T-cell clone with dual specificity for HLA-B27 and HLA-DR2 antigens.

A CD3+ CD4- CD8+ cytolytic T-lymphocyte (CTL) clone, CTL 47, could be induced to proliferate in the presence of exogenous interleukin 2 by either HLA-B27.1+ or HLA-DR2+ cells. B27.1-induced proliferation was strongly and equally inhibited by an anti-B27 and by an anti-CD8 monoclonal antibody (MoAb). DR2-induced proliferation was inhibited by the same anti-CD8 MoAb less efficiently and with a different time course than anti-class II blocking, only being significant when the antibody was added ab initio or very early during the assay. These results indicate that CD8 is essential for class I-induced proliferation but that it also enhances class II-induced stimulation of this CTL clone. It is proposed that the necessary role of CD8 in class I-induced proliferation is related to its interaction with the same class I molecule bound by the T-cell receptor. The accessory role in class II-induced proliferation would be due to an additive effect on the avidity of cell adhesion, resulting from interaction of CD8 with the class I antigens on the stimulator cell, or perhaps to a regulatory role of CD8 as a transducer of early signals for T-cell activation.

Antibodies, Monoclonal↗