RESUMO
BACKGROUND: Control of HIV is suggested to depend on potent effector functions of the virus-specific CD8(+) T-cell response. Antigen opsonization can modulate the capture of antigen, its presentation, and the priming of specific CD8(+) T-cell responses. OBJECTIVE: We have previously shown that opsonization of retroviruses acts as an endogenous adjuvant for dendritic cell (DC)-mediated induction of specific cytotoxic T lymphocytes (CTLs). However, in some HIV-positive subjects, high levels of antibodies and low levels of complement fragments coat the HIV surface. METHODS: Therefore we analyzed the effect of IgG opsonization on the antigen-presenting capacity of DCs by using CD8(+) T-cell proliferation assays after repeated prime boosting, by measuring the antiviral activity against HIV-infected autologous CD4(+) T cells, and by determining IFN-γ secretion from HIV-specific CTL clones. RESULTS: We find that DCs exposed to IgG-opsonized HIV significantly decreased the HIV-specific CD8(+) T-cell response compared with the earlier described efficient CD8(+) T-cell activation induced by DCs loaded with complement-opsonized HIV. DCs exposed to HIV bearing high surface IgG levels after incubation in plasma from HIV-infected subjects acted as weak stimulators for HIV-specific CTL clones. In contrast, HIV opsonized with plasma from patients exhibiting high complement and low IgG deposition on the viral surface favored significantly higher activation of HIV-specific CD8(+) T-cell clones. CONCLUSION: Our ex vivo and in vitro observations provide the first evidence that IgG opsonization of HIV is associated with a decreased CTL-stimulatory capacity of DCs.
Assuntos
Células Dendríticas/imunologia , Infecções por HIV/imunologia , HIV/imunologia , Linfócitos T Citotóxicos/imunologia , Anticorpos Antivirais/imunologia , Apresentação de Antígeno , Antígenos Virais/imunologia , Linfócitos T CD4-Positivos/imunologia , Proliferação de Células , Células Clonais , Proteínas do Sistema Complemento/imunologia , Humanos , Interferon gama/imunologia , Ativação LinfocitáriaRESUMO
BACKGROUND: The complement system is tightly controlled by several regulators. Two of these, factor H (FH) and C4b-binding protein (C4BP), can be acquired by pathogens conveying resistance to complement attack. The aim of the study was to characterize the FH binding molecule of Candida albicans, a potentially life-threatening yeast. METHODS: The gene coding for this molecule was identified by probing an expression library and homozygous deletion mutants of the respective gene were constructed. Binding and functional assays were undertaken to compare wild-type and knockout strains. RESULTS: The high-affinity glucose transporter 1 (CaHgt1p) was identified as an FH-binding molecule. Homozygous hgt1Δ/Δ deletion mutants, but not the restored strain in which HGT1 was reintegrated, showed a decreased binding of FH and even of C4BP, demonstrating its function as an FH- and C4BP-binding protein. This led to an enhanced terminal complement complex deposition after incubation with human serum; CaHgt1p thus functions as complement inhibitor. hgt1Δ/Δ mutants failed to form rosettes with complement-coated sheep erythrocytes, and show reduced binding to HIV-gp160, implying that a complement receptor 3 (CR3) moiety, known as fungal HIV binding molecule is lacking. CONCLUSIONS: CaHgt1p is a multifunctional evasion molecule, as complement inhibitor, CR3 analogue and HIV receptor.
Assuntos
Candida albicans/metabolismo , Candidíase/metabolismo , Complemento C4b/metabolismo , Fator H do Complemento/metabolismo , Complexo de Ataque à Membrana do Sistema Complemento/metabolismo , Proteínas Fúngicas/metabolismo , Proteínas Facilitadoras de Transporte de Glucose/metabolismo , Proteína gp160 do Envelope de HIV/metabolismo , Candida albicans/citologia , Candida albicans/genética , Candida albicans/imunologia , Candidíase/imunologia , Fator H do Complemento/imunologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/imunologia , Proteínas Facilitadoras de Transporte de Glucose/genética , Proteínas Facilitadoras de Transporte de Glucose/imunologia , Humanos , Imunidade Inata , Antígeno de Macrófago 1/metabolismo , Ligação ProteicaRESUMO
Previous studies have demonstrated the involvement of complement (C) in induction of efficient CTL responses against different viral infections, but the exact role of complement in this process has not been determined. We now show that C opsonization of retroviral particles enhances the ability of dendritic cells (DCs) to induce CTL responses both in vitro and in vivo. DCs exposed to C-opsonized HIV in vitro were able to stimulate CTLs to elicit antiviral activity significantly better than non-opsonized HIV. Furthermore, experiments using the Friend virus (FV) mouse model illustrated that the enhancing role of complement on DC-mediated CTL induction also occurred in vivo. Our results indicate that complement serves as natural adjuvant for DC-induced expansion and differentiation of specific CTLs against retroviruses.
Assuntos
Proteínas do Sistema Complemento/imunologia , Células Dendríticas/imunologia , Ativação Linfocitária/imunologia , Infecções por Retroviridae/imunologia , Linfócitos T Citotóxicos/imunologia , Adjuvantes Imunológicos , Animais , Apresentação de Antígeno/imunologia , Feminino , HIV/imunologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos KnockoutRESUMO
BACKGROUND: To determine the correlation between protection and humoral immune response against simian immunodeficiency virus (SIVmac251), 11 macaques were immunized with live-attenuated SIVmac239Deltanef either intravenously or via the tonsils and exposed to SIVmac251 after either 6 or 15 months along with unvaccinated controls. RESULTS: Independent of the route of vaccine application, viremia was significantly reduced in vaccinees compared with controls 2 weeks post-challenge. Concomitantly, viremia correlated inversely with SIV-specific IgG, complement-mediated lysis and neutralizing antibodies and these parameters seemed to contribute to reduced viremia. During chronic infection, six monkeys controlled viremia in the circulation (two or fewer infectious units per 10(6) PBMCs) and showed no signs of trapping in lymphatic tissues (Appendix S1). CONCLUSIONS: As no significant differences were observed throughout the study, with respect to the humoral immune response and viremia control, between the two vaccinated cohorts, mucosal immunization strategies are recommended due to more simplified application.
Assuntos
Macaca mulatta/imunologia , Vacinas contra a SAIDS/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/imunologia , Vacinação/veterinária , Administração Sublingual , Animais , Anticorpos Antivirais/sangue , Estudos de Coortes , Citometria de Fluxo/veterinária , Imunidade Humoral/imunologia , Hibridização In Situ/veterinária , Injeções Intravenosas/veterinária , Macaca mulatta/virologia , Testes de Neutralização/veterinária , RNA Viral/sangue , Vacinas contra a SAIDS/administração & dosagem , Síndrome de Imunodeficiência Adquirida dos Símios/prevenção & controle , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Estatísticas não Paramétricas , Vacinação/métodos , Vacinas Atenuadas/administração & dosagem , Vacinas Atenuadas/imunologia , Viremia/imunologia , Viremia/veterinária , Viremia/virologiaRESUMO
HLA-specific antibodies generated by allo-immunization are supposed to be involved in the control of HIV infections by both the neutralizing capacity of HLA-specific antibodies (Abs) and HLA-specific Ab-dependent complement-mediated lysis (CML). We further characterized CML of HIV primary isolates induced by HLA-specific Abs. Although HIV-specific and HLA allo-type specific Abs induced only weak CML of HIV primary isolates, several combinations of HLA allo-type specific Abs with HIV-specific Abs could enhance CML significantly. Nevertheless, certain HLA-specific Abs did not improve but even inhibit CML of HIV, although the corresponding HLA molecules were present. Thus, our results emphasize a possible limitation of allo-immunization as a potential approach to induce protective immunity against HIV.
Assuntos
Proteínas do Sistema Complemento/imunologia , Anticorpos Anti-HIV/imunologia , Infecções por HIV/imunologia , HIV-1/imunologia , Antígenos HLA/imunologia , Animais , Infecções por HIV/terapia , Infecções por HIV/virologia , Humanos , Alótipos de Imunoglobulina/imunologia , Imunoterapia/métodos , CamundongosRESUMO
Zygomycosis is increasingly recognized in immunocompromised hosts. We investigated whether platelets become activated after contact with Zygomycetes and adhere to conidial and hyphal structures using immunofluorescence. The platelets' influence on fungal viability was evaluated by assessing hyphal elongation and hyphal damage. Platelets became activated and strongly adhered to conidia and hyphae of Zygomycetes. Platelets induced time dependent damage to hyphae and significantly reduced (P<.05) hyphal elongation. We found that platelets possess antifungal capacities against Zygomycetes based on granule dependent mechanisms and significantly reduce fungal growth and spread, both of which are of major importance in evolving invasive disease.
Assuntos
Plaquetas/imunologia , Mucorales/imunologia , Ativação Plaquetária/imunologia , Adesão Celular , Células Cultivadas , HumanosRESUMO
Infections with enterohemorrhagic Escherichia coli (EHEC) are a major cause of hemolytic uremic syndrome (HUS). Shiga toxins (Stxs), especially Stx2, are believed to represent major virulence factors of EHEC, contributing to HUS pathogenesis. Beside EHEC-associated HUS, there are hereditary atypical forms of HUS, which are mostly caused by mutations of complement regulators. The aim of the present study was to investigate whether or not complement is also involved in the pathogenesis of EHEC-induced typical HUS, by being activated either directly or indirectly by involvement of its inhibitors. Purified Stx2 markedly activated complement via the alternative pathway and was found to bind to factor H (FH), however, only when it was active. No apparent cleavage or destruction of FH was visible, and cofactor activity in fluid phase was unaffected, but clearly delayed for surface-attached FH, where it is essential for host cell protection. Binding studies using FH constructs revealed that Stx2 binds to short consensus repeats (SCRs) 6-8 and SCRs18-20, but not to SCRs16-17, i.e., to regions involved in the surface recognition function of FH. In conclusion, complement, and in particular FH, not only plays an important role in atypical HUS, but most probably also in EHEC-induced HUS.
Assuntos
Ativação do Complemento/fisiologia , Fator H do Complemento/metabolismo , Síndrome Hemolítico-Urêmica/metabolismo , Toxina Shiga/metabolismo , Animais , Western Blotting , Células CHO , Cricetinae , Cricetulus , Escherichia coli Êntero-Hemorrágica , Ensaio de Imunoadsorção Enzimática , Infecções por Escherichia coli/complicações , Síndrome Hemolítico-Urêmica/microbiologia , Humanos , ImunoprecipitaçãoRESUMO
Besides Shiga toxins (Stx), Stx-producing Escherichia coli (STEC) harbour several other putative virulence factors, including the serine protease EspP. We have investigated 214 STEC strains from Austria belonging to 61 different serotypes from humans, animals, and food for the presence of this serine protease gene and have determined the espP subtypes and their association with clinical outcome. espP was detected in 121 (57%) out of 214 strains. Sixty-five of 68 strains (96%) of non-sorbitol-fermenting (NSF) O157:H7/NM (NM, non-motile) were positive for espP, while none of 8 SF E. coli O157:NM isolates contained this gene. All 9 strains of serotype O145:NM and 17 of 21 strains (81%) of serotype O26:H11/NM were positive for espP. Nineteen STEC serogroups including O103 and O111 serogroups--considered to be highly pathogenic--were completely negative for espP. Only 5 of 12 strains isolated from patients suffering from haemolytic uraemic syndrome (HUS) were espP-positive (all serogroup NSF O157) as well as 28 of 39 strains from patients with bloody diarrhoea, 40 of 63 strains from patients with non-bloody diarrhoea, and 15 of 19 strains from asymptomatic patients. In O157:H7/NM, O26:H11/NM, and O145:NM only espP subtype alpha was found, whereas in most of the other non-O157 serogroups, subtypes beta and gamma were found. Subtype delta was not detected in our strain collection. Regarding the espP subtypes, only subtype alpha, but not beta and gamma, were found in HUS patients. Moreover, we could demonstrate that espP, and in particular subtype alpha, is associated with highly pathogenic serogroups.
Assuntos
Proteínas de Escherichia coli/genética , Serina Endopeptidases/genética , Escherichia coli Shiga Toxigênica/enzimologia , Escherichia coli Shiga Toxigênica/patogenicidade , Fatores de Virulência/genética , Animais , Áustria , DNA Bacteriano/genética , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/veterinária , Microbiologia de Alimentos , Genótipo , Humanos , Reação em Cadeia da Polimerase/métodos , Sorotipagem , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/isolamento & purificaçãoRESUMO
Cerebral aspergillosis is a mostly lethal infection of the central nervous system. Former results identified low cerebral complement levels as one cause for insufficient immune reaction. Therefore we studied cerebral complement expression after fungal invasion and investigated putative mechanisms of Aspergillus spp to cope with the complement-induced selection pressure. Brain tissue derived from patients with cerebral aspergillosis or non-infected individuals was analyzed immunohistochemically for complement synthesis. Correlations between expression levels were determined statistically. Increased complement synthesis, a prerequisite for strengthened antifungal potency, was visible in resident astrocytes, neurons, oligodendrocytes as well as in infiltrating macrophages after fungal infection. Surprisingly, microglia, although regarded as major immune cells, only marginally participated in synthesis of most complement proteins. Several evasion mechanisms were found that help the fungus to establish a cerebral infection even in the presence of complement: Fungal hyphae limit the surface deposition of C3 and thus interfere with complement-dependent phagocytosis. Furthermore, the "sealing off" in brain abscesses not only inhibits fungal spreading but also forms protection shields against complement attack. Complement indeed seems to represent an important selection pressure and evokes the development of fungal evasion mechanisms. Counteractions for these evasion processes might represent interesting therapeutic approaches.
Assuntos
Aspergillus/imunologia , Encefalopatias/imunologia , Encefalopatias/microbiologia , Proteínas do Sistema Complemento/imunologia , Neuroaspergilose/imunologia , Adulto , Idoso , Encéfalo/patologia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-IdadeRESUMO
Using laser scanning microscopy, we investigated whether platelets are capable of internalizing Aspergillus conidia and examined Aspergillus-platelet adherence. The influence of platelets on fungal growth was evaluated by assessing galactomannan (GM) release, hyphal elongation, and colony size. A secretion assay with [(3)H]-serotonin (5-hydroxytryptamine [5-HT]) was performed. Exposure to platelets resulted in significantly decreased GM release (p<.05), hyphal elongation (p<.001), colony size, pigmentation, and 5-HT release ( p<.05). A lack of antifungal effects was observed with the microfilament inhibitor cytochalasin D. Platelets attenuate the virulence of Aspergillus species in vitro on the basis of granule-dependent effects.
Assuntos
Aspergillus fumigatus/patogenicidade , Aspergillus/patogenicidade , Plaquetas/fisiologia , Plaquetas/ultraestrutura , Grânulos Citoplasmáticos/metabolismo , Aspergillus/classificação , Aspergillus/metabolismo , Aspergillus/fisiologia , Aspergillus fumigatus/metabolismo , Aspergillus fumigatus/fisiologia , Adesão Celular , Galactose/análogos & derivados , Humanos , Mananas/metabolismo , Microscopia Confocal , Microscopia de Fluorescência , Serotonina/metabolismo , Serotonina/farmacologiaRESUMO
The in vitro activity of posaconazole plus amphotericin B against conidia and hyphae of 30 clinical zygomycetes was investigated. The combination of posaconazole with amphotericin B was found to be significantly more synergistic (40%) against hyphae (P < 0.05) than against conidia (10%). Antagonism was not observed.
Assuntos
Anfotericina B/administração & dosagem , Antifúngicos/administração & dosagem , Mucorales/efeitos dos fármacos , Triazóis/administração & dosagem , Absidia/efeitos dos fármacos , Cunninghamella/efeitos dos fármacos , Sinergismo Farmacológico , Humanos , Hifas/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Mucor/efeitos dos fármacos , Mucorales/patogenicidade , Mucorales/ultraestrutura , Mucormicose/tratamento farmacológico , Mucormicose/microbiologia , Rhizomucor/efeitos dos fármacos , Rhizopus/efeitos dos fármacos , Esporos Fúngicos/efeitos dos fármacosRESUMO
Secreted aspartic proteases (Saps) represent an important virulence factor facilitating fungal adherence. Several protease inhibitors (PIs), including HIV PIs, have been shown to reduce Candida adhesion. The aim of this study was to ascertain whether or not the recently discovered PIs Aureoquinone and Laccaridiones A and B, isolated from Basidiomycete cultures, or Bestatin, act as Sap-inhibitors and/or inhibitors of fungal adhesion. Drug effects on candidial Sap-production were determined by Sap-ELISA. Control tubes, in the absence of drug, served as positive controls, while tubes excluding both drug and proteinase induction medium were used as negative controls. Aureoquinone as well as Laccaridiones A and B, but not Bestatin, significantly inhibited Candida albicans adhesion to both epithelial and endothelial cells in a dose dependent manner and also reduced Sap-release (effects were not because of a direct interaction of the Basidiomycete metabolites with secreted Saps). Laccaridione B was consistently found to be the most effective PI tested. Interestingly, these drugs are neither fungistatic nor fungicidal at the concentrations applied. Laccaridione B may represent a promising novel type of antimycotic drug--targeting virulence factors without killing the yeast.
Assuntos
Antibiose , Basidiomycota/metabolismo , Candida albicans/patogenicidade , Inibidores de Proteases/farmacologia , Benzoquinonas/metabolismo , Candida albicans/crescimento & desenvolvimento , Adesão Celular , Linhagem Celular , Células Endoteliais/microbiologia , Células Epiteliais/microbiologia , Humanos , Leucina/análogos & derivados , Leucina/metabolismo , Quinonas/metabolismoRESUMO
MICs and fractional inhibitory concentrations were evaluated for anidulafungin and voriconazole alone and in combination against conidia and hyphae under hypoxic (1% oxygen-5% CO(2)-94% nitrogen) conditions against 31 Aspergillus isolates. Anidulafungin exhibited excellent activity against conidia and hyphae of Aspergillus spp. The visual reading of the MIC for anidulafungin was optimal under hypoxic conditions.
Assuntos
Aspergillus/efeitos dos fármacos , Equinocandinas/farmacologia , Hifas/efeitos dos fármacos , Pirimidinas/farmacologia , Esporos Fúngicos/efeitos dos fármacos , Triazóis/farmacologia , Anaerobiose , Anidulafungina , Antifúngicos/farmacologia , Aspergillus fumigatus/efeitos dos fármacos , Sinergismo Farmacológico , Testes de Sensibilidade Microbiana , VoriconazolRESUMO
This study investigated the basis for intrinsic amphotericin B (AMB) resistance in Aspergillus terreus. The ergosterol content, cell wall composition, and lipid peroxidation level had no influence on AMB resistance. The level of catalase production in A. terreus was significantly higher than that in A. fumigatus (P < 0.05). This higher-level production may contribute to AMB resistance in A. terreus since oxidative damage has been implicated in AMB action.
Assuntos
Anfotericina B/farmacologia , Antifúngicos/farmacologia , Aspergillus/classificação , Aspergillus/efeitos dos fármacos , Catalase/biossíntese , Farmacorresistência Fúngica , Aspergilose , Aspergillus/enzimologia , Humanos , Testes de Sensibilidade MicrobianaRESUMO
Upon crossing the endothelial barrier of the host, HIV initiates immediate responses of the immunity system. Among its components, the complement system is one of the first the first elements, which are activated to affect HIV propagation. Complement participates not only in the early phase of the immune response, but its effects can be observed continuously and also concern the induction and modification of the adaptive immune response. Here we discuss the role of complement in early and late stages of HIV pathogenesis and review the escape mechanisms, which protect HIV from destruction by the complement system.
Assuntos
Ativação do Complemento , Proteínas do Sistema Complemento/imunologia , Infecções por HIV/imunologia , HIV/imunologia , Anafilatoxinas/imunologia , Animais , Células Dendríticas/imunologia , Células Dendríticas/virologia , Infecções por HIV/sangue , Interações Hospedeiro-Patógeno , Humanos , Tecido Linfoide/imunologia , Tecido Linfoide/virologia , Mucosa/imunologia , Mucosa/virologia , Proteínas Opsonizantes/imunologia , Ligação ViralRESUMO
Literature harbours several reports of potable water-associated outbreaks. We studied the prevalence of Shiga toxin- (stx1/2), intimin- (eae) and haemolysin (hlyA) genes in Escherichia coli isolates from drinking water of private and public water supplies in a rural area of Upper Austria; 2633 water samples were gained between November 2000 and December 2003. Two hundred and eighty of these water samples were positive for E. coli (10.6%). Of these, 101 samples were drawn from drilled wells (36%), 96 from dug wells (34%), 61 from springs (22%) and 22 from water supplies without available information on technical details (8%); 141 of the samples were from public water supplies, 139 from private water supplies. Eleven of the E. coli isolates were found to be positive for one of the investigated virulence genes (3.9%): one isolate yielded stx2, seven eae, and three isolates had hlyA. The presence of these genes underlines the importance of control of water quality in public and also private water supplies.
Assuntos
Adesinas Bacterianas/isolamento & purificação , Escherichia coli Enteropatogênica/genética , Proteínas de Escherichia coli/isolamento & purificação , Proteínas Hemolisinas/isolamento & purificação , Toxina Shiga I/isolamento & purificação , Toxina Shiga II/isolamento & purificação , Microbiologia da Água , Abastecimento de Água/análise , Adesinas Bacterianas/genética , Áustria , Ingestão de Líquidos , Eletroforese em Gel de Campo Pulsado , Escherichia coli Enteropatogênica/isolamento & purificação , Escherichia coli O157/genética , Proteínas de Escherichia coli/genética , Proteínas Hemolisinas/genética , Humanos , Reação em Cadeia da Polimerase , Prevalência , Estudos Prospectivos , População Rural , Toxina Shiga I/genética , Toxina Shiga II/genética , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/isolamento & purificaçãoRESUMO
Due to ongoing recombination and mutations, HIV permanently escapes from neutralizing antibody (nAb) responses of the host. By the masking of epitopes or shedding of gp120, HIV-1 further impedes an efficient neutralization by Abs. Therefore, nAbs responses of the host are chasing behind a rapidly evolving virus and mainly non-neutralizing antibodies (non-nAbs) are present in the host. At the same time, complement deposition on immune-complexed HIV may counteract the immune response by enhancing the infection. On the other hand, complement-mediated lysis is a putative effector mechanism to control viral replication. Here we review the complex interplay between complement, neutralizing and non-neutralizing Abs during HIV infection and discuss the contribution of Abs and complement in blocking versus enhancing the course of infection.
Assuntos
Proteínas do Sistema Complemento/fisiologia , Anticorpos Anti-HIV/fisiologia , Infecções por HIV/imunologia , Citotoxicidade Celular Dependente de Anticorpos , Linfócitos B/fisiologia , Células Dendríticas Foliculares/fisiologia , Eritrócitos/fisiologia , Humanos , Integrina alfaXbeta2/fisiologia , Antígeno de Macrófago 1/fisiologiaRESUMO
Shiga toxins (Stx) are believed to play a key role in the pathogenesis of diseases caused by Stx-producing Escherichia coli (STEC), including the potentially life-threatening hemolytic uremic syndrome (HUS). In this study, 201 STEC strains collected from patients and environmental sources were investigated with regard to the stx genotypes and pathogenicity. The stx(2) and stx(2c) alleles were associated with high virulence and the ability to cause HUS, whereas stx(2d), stx(2e,)stx(1), and stx(1c) occurred in milder or asymptomatic infections. Quantification of Stx using an enzyme immunoassay and the Vero cell cytotoxicity assay showed no significant differences between the strains associated with HUS and those causing milder diseases. We hypothesize that the stx genotype and perhaps other yet unknown virulence factors rather than the amount of Stx or the in vitro cytotoxicity correlate with the development of HUS.
Assuntos
Síndrome Hemolítico-Urêmica/microbiologia , Toxina Shiga I/genética , Toxina Shiga II/genética , Escherichia coli Shiga Toxigênica/genética , Adesinas Bacterianas/metabolismo , Animais , Chlorocebus aethiops , Proteínas de Escherichia coli/metabolismo , Microbiologia de Alimentos , Doenças Transmitidas por Alimentos , Genótipo , Humanos , Técnicas Imunoenzimáticas , Toxina Shiga I/classificação , Toxina Shiga II/classificação , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/patogenicidade , Células Vero/microbiologia , Virulência/genéticaRESUMO
Outbreaks traced to bacterial contamination of multiple-dose vials are reported in the literature. During a four-month period, multi-dose vials (MDVs), single-dose vials (SDVs), and vials containing self-prepared admixtures were collected from various wards to analyse sterility of their contents. We examined 68 commercially available MDVs containing sodium chloride 0.9% or heparin with added preservative and 17 single dose vials (SDVs) containing aqua ad injectionem or sodium chloride 0.9% and 11 vials with admixtures (ADX) of heparin and sodium chloride 0.9%, both without preservative. Four of 96 (4.17%) vials were not sterile: two of them were contaminated with spore-forming bacteria, two with coagulase negative Staphylococci. Three of the samples were MDVs containing a preservative. The date of the first use was not marked on 28% of the vials. Twenty-eight samples were multiply used, although they were SDVs or ADXs without preservative or without an adequate amount of preservative, respectively. In 15 of 68 MDVs, the time limit after the first use was exceeded. On average, the volume of the samples was 80% of the original volume. A proportion of 4% of vials was not sterile. A training programme for health care workers in aseptic techniques and for validation of the preparation of solutions for parenteral use should be installed.