Flow cytometric quantitation of phagocytosis in heparinized complete blood with latex particles and Candida albicans

Egido, J.; Viñuelas, Juan

doi:10.1590/s0037-86821997000600001

Cited by 6 publications

(5 citation statements)

References 12 publications

(5 reference statements)

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“…A 250‐μL aliquot of serum preincubated Candida cell suspension was mixed with 50 μL of PBLs (1 × 10 7 cells mL −1 ) to obtain a target cell : effector ratio of 4 : 1, and incubated for the next 30 min at 37 °C. After lysis of remaining red blood cells with ammonium chloride, the pellet was washed twice with PBS (1650 g , 10 min, 4 °C) and resuspended with 500 μL of lysis buffer (2.5% sodium desoxycholate, pH 8.7) for 10 min to lyse PBLs (Egido & Viñuelas, 1997; Soloviev et al , 2007). Cells were washed twice with PBS, incubated with 10 μL of RNAse (1 mg mL −1 ) for 30 min at room temperature, and then washed twice with PBS.…”

Section: Methodsmentioning

confidence: 99%

Model α-mannoside conjugates: immunogenicity and induction of candidacidal activity

Paulovičová

Bystrický

Paulovičová

et al. 2010

FEMS Immunol Med Microbiol

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The effect of Candida cell wall mannan-derived alpha-oligomannoside structural components on the modulation of the immune system and their role in protective immunity are studied here. Semi-synthetic alpha-mannoside-bovine serum albumin conjugates were used for immunization of rabbits. Dimeric alpha-mannoside, representing Candida antigenic factor 1, was used as a model of linear alpha-mannoside, and pentameric alpha-mannoside was used as a model of branched oligomannoside side chain structure. The induction of humoral immune response and the functionality of the serum tested by induction of peripheral blood leukocyte (PBL) candidacidal activity are documented. Anti-Candida albicans serotype B immunoglobulins (IgG and IgM) levels were higher than anti-serotype A following immunization with both conjugates. Dimer-conjugate postimmunization sera evidently enhanced C. albicans killing activity of PBLs in candidacidal assay. The study shows the importance of alpha-mannoside structures in perspective anti-Candida vaccine with a broad spectrum of effectiveness.

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Section: Methodsmentioning

confidence: 99%

Model α-mannoside conjugates: immunogenicity and induction of candidacidal activity

Paulovičová

Bystrický

Paulovičová

et al. 2010

FEMS Immunol Med Microbiol

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“…The development of fluorochromes to detect oxidative bursts due to phagocytosis (17,251,281) increased the number of studies with different microbes such as Borrelia burgdorferi (17), Staphylococcus spp. (128,196), Escherichia coli (70,271), Bordetella pertussis (299), Cryptococcus neoformans (48), Salmonella (272), and yeasts (90,96,114).…”

Section: General Applications Of Flow Cytometry To Microbiologymentioning

confidence: 99%

“…Recently, Peyron et al (252) have compared FCM with the NCCLS broth dilution method for amphotericin B. Oxonol was used to measure the variations in membrane potential. From the dose-response curve obtained for each isolate, the concentrations at which the fluorescence intensity was reduced by 50, 80, and 90%, i.e., the IC 50 , IC 80 , and IC 90 , respectively, were calculated. Regression analysis revealed that the best agreement with the M27-T NCCLS document was obtained with the IC 80 end point.…”

Section: Antifungal Agentsmentioning

confidence: 99%

Applications of Flow Cytometry to Clinical Microbiology

Alvarez-Barrientos¹,

Arroyo²,

Cantón³

et al. 2000

Clinical Microbiology Reviews

156

111

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Section: General Applications Of Flow Cytometry To Microbiologymentioning

confidence: 99%

Applications of Flow Cytometry to Clinical Microbiology

Álvarez-Barrientos¹,

Arroyo

Cantón³

et al. 2000

Clin Microbiol Rev

182

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SUMMARY Classical microbiology techniques are relatively slow in comparison to other analytical techniques, in many cases due to the need to culture the microorganisms. Furthermore, classical approaches are difficult with unculturable microorganisms. More recently, the emergence of molecular biology techniques, particularly those on antibodies and nucleic acid probes combined with amplification techniques, has provided speediness and specificity to microbiological diagnosis. Flow cytometry (FCM) allows single- or multiple-microbe detection in clinical samples in an easy, reliable, and fast way. Microbes can be identified on the basis of their peculiar cytometric parameters or by means of certain fluorochromes that can be used either independently or bound to specific antibodies or oligonucleotides. FCM has permitted the development of quantitative procedures to assess antimicrobial susceptibility and drug cytotoxicity in a rapid, accurate, and highly reproducible way. Furthermore, this technique allows the monitoring of in vitro antimicrobial activity and of antimicrobial treatments ex vivo. The most outstanding contribution of FCM is the possibility of detecting the presence of heterogeneous populations with different responses to antimicrobial treatments. Despite these advantages, the application of FCM in clinical microbiology is not yet widespread, probably due to the lack of access to flow cytometers or the lack of knowledge about the potential of this technique. One of the goals of this review is to attempt to mitigate this latter circumstance. We are convinced that in the near future, the availability of commercial kits should increase the use of this technique in the clinical microbiology laboratory.

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Flow cytometric quantitation of phagocytosis in heparinized complete blood with latex particles and Candida albicans

Cited by 6 publications

References 12 publications

Model α-mannoside conjugates: immunogenicity and induction of candidacidal activity

Model α-mannoside conjugates: immunogenicity and induction of candidacidal activity

Applications of Flow Cytometry to Clinical Microbiology

Applications of Flow Cytometry to Clinical Microbiology

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