Biomedical subjects

D Saverino

Publications and source records attributed to D Saverino.

26 records · Page 2Linked to original sources

PubMed1995

Recognition of human T-leukemia virus (HTLV-1) envelope by human CD4+ T-cell lines from HTLV-1 seronegative individuals: specificity and clonal heterogeneity.

Because T-helper cells are critical for immune responses in retroviral infections, CD4+ T-cell lines specific for the human T-leukemia virus type 1 (HTLV-1) envelope have been generated from peripheral T lymphocytes of nonimmune donors to study their naive repertoire. Recombinant fragments (RE1, amino acids [aa] 26-200; RE3, aa 165-307; RE5, aa 308-401; and RE6, aa 165-401) of HTLV-1 envelope, whole envelope glycoprotein, and synthetic peptides were used to induce T-cell lines. CD4+ T-cell lines specific for one or more fragments were obtained from seven of eight individuals tested. T-cell lines generated against envelope glycoprotein from five of five donors did not cross-react with the RE fragments and vice versa. The lines specific for RE and env were mapped with overlapping peptides. The lines with single peptide (narrow) specificity contained a variety of clones that used different T-cell receptor V beta genes. These data (1) suggest that most of the normal individuals carry T-helper precursors specific for epitopes on HTLV-1 envelope; (2) indicate that heterogeneity of HTLV-1 envelope-specific T cells can be detected in the naive repertoire; and (3) define optimal antigenic preparations to be used to assess cellular immunity in HTLV-1-infected individuals.

CD4-Positive T-Lymphocytes↗

PubMed1995

Human T helper cells specific for HIV reverse transcriptase: possible role in intrastructural help for HIV envelope-specific antibodies.

Cooperation between B cells specific for an antigen exposed on a viral structure and T helper (Th) cells specific for an internal antigen, as demonstrated with influenza, hepatitis B and rabies viruses, has been termed intrastructural help. Th cells specific for internal proteins of HIV, which are much less mutated than its exposed antigens, may be valuable in vaccine design against this virus. We investigated the human Th repertoire specific for the core HIV antigen reverse transcriptase (p66), and determined whether these cells could be candidate intrastructural T helpers. CD4+ T lines and clones were generated from non-immune individuals by stimulation with p66-pulsed antigen-presenting cells (APC). Specific lines were obtained with p66 from 19 out of 21 (90%) of these individuals, vs. 7 out of 29 (24%) with gp120. Diverse epitopes were recognized by different individuals, and various V beta genes were used by these clones. Clones using the same V beta genes were of diverse origin, according to VDJ region sequence. Of these lines 45% responded to p66 in the context of HIV virions. Moreover, p66-specific clones could respond to APC that had internalized HIV complexed with envelope-specific monoclonal antibodies, suggesting that p66-specific Th cells may participate in intrastructural help. These studies indicate that p66-specific Th cells are detectable in vitro in most naive individuals and exhibit clonal heterogeneity, and that the majority recognize an HIV conserved antigen. They respond to p66 following processing of whole virions and are clearly candidates for intrastructural help. If confirmed in vivo, p66 should be included among vaccine candidates investigated to optimize the anti-HIV Th response.

Amino Acid Sequence↗

PubMed1995

Antistaphylococcal activity of cefdinir, a new oral third-generation cephalosporin, alone and in combination with other antibiotics, at supra- and sub-MIC levels.

Cefdinir is one of the few oral third generation cephalosporins that shows useful activity against nosocomial Gram-positive pathogens. For this reason the anti-staphylococcal potency of the new drug, alone or in combination with other drugs was further characterized. Against penicillin-resistant, oxacillin-susceptible Staphylococcus isolates, cefdinir demonstrated useful in-vitro activity. MIC90 values (in mg/L) were 0.25 for Staphylococcus aureus (30 strains), 0.06 for Staphylococcus epidermidis (24), 0.125 for Staphylococcus hominis (10), 0.5 for both Staphylococcus xylosus (15) and Staphylococcus capitis (11) and 4 for Staphylococcus saprophyticus (10), while Staphylococcus haemolyticus (12) was less susceptible with a MIC90 value of 32. Cefdinir activity was not adversely affected by several variables such as pH, inoculum size or the presence of serum or urine. The new cephem induced a PAE on all isolates studied: with S. aureus the extent of regrowth suppression ranged from 0.8 to 1 h, and with the other species from 0.5 (S. epidermidis) to 4.1 h (S. haemolyticus). Development of resistant strains was rare. At the highest level used (10 x MIC) mutants arose with a frequency of 6 x 10(-8) with S. haemolyticus and 2 x 10(-9) with S. epidermidis. The absence of a paradoxical effect of increasing concentrations of cefdinir on its bactericidal activity was confirmed up to a value of 500-fold the MICs. When cefdinir activity was assessed in association with ciprofloxacin, netilmicin, clarithromycin, fosfomycin, rifampicin, teicoplanin and vancomycin using the chequerboard and time-kill techniques, indifference predominated with all strains and in all combinations. Synergism was detected only in 11 out of a total of 175 tests performed by the chequerboard method. Using the time-kill technique cefdinir reacted synergically in 25 of 126 tests. Antagonism was never observed. S. aureus exposed to sub-inhibitory concentrations of cefdinir failed to grow on mannitol-salt agar and to produce haemolysins, but retained coagulase activity. Penicillinase production was also lost in about 17% of the survivors. Hydrophobicity changes were detected in all species tested with the exception of S. saprophyticus.

Anti-Bacterial Agents↗

PubMed1993

Plasmid loss from gram-negative bacteria exposed to subinhibitory concentrations of beta-lactam drugs and azithromycin.

The stability of F'lac, pW101 and pHSG298 in Escherichia coli K12 exposed to subinhibitory concentrations of beta-lactam antibiotics, amikacin and tetracycline was studied. High molecular weight low copy plasmids (F'lac and pW101) were eliminated from bacteria treated with PBP-3 binding molecules, while a low molecular weight high copy extrachromosomal element (pHSG298) was not. None of the carbapenem antibiotics, mecillinam, amikacin or tetracycline promoted high rate plasmid loss from their hosts. Under the same conditions, plasmid-mediated ampicillin-resistance due to beta-lactamase production was also lost from F'lacTn1-carrying bacteria. In contrast, the high copy R6K plasmid was stably inherited in their hosts with the exception of those organisms treated with cefixime. When the same experiments were performed with a Klebsiella pneumoniae strain induced to form filaments by azithromycin at sub-MICs, F'lacTn1 and pW101 loss was detected, while pHSG298 was stably inherited. These results confirm previous observations that plasmid stability is correlated with cell shape and that recovery is more easily achieved when bacteria undergo an unbalanced division resulting in cell filamentation.

Anti-Bacterial Agents↗

PubMed1992

Parameters characterizing the in vitro activity of cefixime, a new oral broad spectrum cephalosporin, against respiratory and urinary pathogens.

The wide and potent in vitro activity of cefixime, a new oral broad spectrum cephalosporin, has been confirmed on a collection of respiratory and urinary pathogens recently isolated in Italy. The new cephem emerged as the most bactericidal of all the comparators tested against several fast as well as slowly-growing gram-negative species including Enterobacteria, Haemophilus and Moraxella, irrespective of their ability to synthetize beta-lactamases. Among the gram-positive species Streptococcus pyogenes and S. pneumoniae were effectively covered. Cefixime activity was not adversely influenced by several important variables such as pH (over the range from 5 to 8), inoculum size (from 10(5) to 10(8) CFU per ml) and the presence of 50% human serum or urine. Time-kill tests confirmed a pronounced bactericidal potency of the drug especially towards common respiratory pathogens (H. influenzae, M. catarrhalis, S. pneumoniae and S. pyogenes). Killing of urinary strains was optimal at cefixime concentrations reached in urine since eradication, except for Proteus mirabilis, was enhanced with increasing levels of the drug. The absence of an untoward paradoxic effect on the rate of cefixime bactericidal action was confirmed by employing a dynamic bladder model simulating the pharmacokinetic parameters of the drug after a single 200 mg daily dosage. Interactions of cefixime with several other drugs that may be employed in combination therapy were generally prone to provide indifference and synergism while antagonism was never observed. Favorable interactions were also registered when cefixime acted with other antibiotics on partially resistant species such as Staphylococci and Pseudomonas. The new cephem seems to provide excellent opportunities for expanding oral cephalosporin therapy to a wide range of infections produced by susceptible pathogens in the adult and pediatric populations.

Anti-Infective Agents, Urinary↗

PubMed1992

Antibacterial profile of flurithromycin, a new macrolide.

The in-vitro activity of flurithromycin against common respiratory tract Gram-positive (85 strains) and Gram-negative (44 strains) pathogens, and a collection of anaerobes (125 strains) was compared with that of erythromycin, cefixime, amoxycillin, co-amoxiclav, ciprofloxacin, netilmicin and clindamycin. Flurithromycin possessed the same spectrum and potency of antimicrobial activity as erythromycin. The presence of 50% human serum in the test media enhanced the activity of flurithromycin against all isolates, with the exception of Streptococcus pyogenes. Flurithromycin induced a post-antibiotic effect (PAE) that ranged from 0.25 h (Haemophilus influenzae) to 3.5 h (Streptococcus pneumoniae) for all strains tested. The presence of serum increased or diminished the duration of the PAE, depending on the strain being analysed. No interaction between flurithromycin and the other drugs tested was observed by the checkerboard technique, but when the time-kill system was used, 35 cases of synergy were noted out of 120 tests performed (29%), of which 15 (43%) were with Moraxella catarrhalis, 12 (34%) with Staphylococcus aureus, four (11%) with H. influenzae and the remainder with S. pneumoniae and S. pyogenes. Netilmicin produced most synergic interactions. Antagonism was not detected by either methods.

Bacteria, Anaerobic↗

PubMed1991

Synergistic antibacterial interaction of cefotaxime and desacetylcefotaxime.

Cefotaxime (CTX) is metabolized in desacetylcefotaxime (dCTX), a less potent compound which shows, however, a higher stability against selected beta-lactamases produced by Gram-negative organisms. The aim of this study was to verify if the antimicrobial activity of CTX against 260 clinical aerobic and anaerobic pathogens isolated in our institution was enhanced by its metabolic derivative dCTX. The combination of CTX and dCTX, assessed by checkerboard titration, was completely or partially synergistic towards 61% of the 220 aerobic organisms tested and against 68% of the 40 Bacteroides strains analyzed. In addition we have investigated, by the time-kill method, the in-vitro interactions against 50 aerobic strains of CTX and dCTX alone and in combination with netilmicin, a drug often employed in severe infections in combination with beta-lactam agents in order to provide effective killing of resistant nosocomial pathogens. Time-kill studies indicated that 36% of the aerobic nosocomial strains were synergistically inhibited by the combination of CTX/dCTX with netilmicin. These results indicate that dCTX makes an important contribution to the clinical efficacy of CTX.

Bacteria↗

PubMedDate not supplied

Handling of retroviral antigens by human antigen-presenting cells.

Antigen-specific T helper cells play an important role in retroviral infections. Indeed, they provide help for B-cell activation and antibody production and for clonal expansion of cytolytic lymphocytes. Therefore, we used retrovirus-specific human T helper clones in order to define modes of antigen presentation, antigen-presenting cells and the molecular context of Th epitopes that could be exploited in the design of immunogens aimed at optimizing the Th cell response. In particular, we describe several mechanisms of receptor-mediated antigen uptake that enhance the stimulation of human T-cell clones specific for HIV and HTLV-1 antigens; we report on the differential recognition of Th epitopes depending on the molecular-viral context; we show that dendritic cells are the most efficient presenting cells and are essential for the induction of in vitro primary Th cell responses; and finally, we propose that Th cells specific for internal, conserved antigens of HIV such as reverse transcriptase, may be candidates for intrastructural help resulting in induction of envelope specific antibodies.

Antigen Presentation↗