scholarly journals Quantification of Human β-Defensin-2 and -3 in Body Fluids: Application for Studies of Innate Immunity

2007 ◽  
Vol 53 (4) ◽  
pp. 757-765 ◽  
Author(s):  
Santosh K Ghosh ◽  
Thomas A Gerken ◽  
Keith M Schneider ◽  
Zhimin Feng ◽  
Thomas S McCormick ◽  
...  
Keyword(s):  
Total Protein ◽  
Body Fluids ◽  
Cationic Peptides ◽  
Healthy Donors ◽  
Suction Blister Fluid ◽  
Vaginal Swabs ◽  
Health And Disease ◽  
Cervicovaginal Lavage ◽  
Healthy Participants ◽  
Assay Conditions

AbstractBackground: Human β-defensins (hBDs) are epithelial cell-derived antimicrobial and immunoregulatory cationic peptides. Our objective was to establish an analytical tool to quantify inducible hBD-2 and -3 in body fluids.Methods: We developed sandwich ELISAs using commercially available capture and detection antibodies and determined optimal assay conditions (with 250 mmol/L CaCl2) to overcome masking by endogenous components of body fluids. We used recombinant hBD as calibrators and for recovery testing.Results: hBD-2 and -3 detection limits were ∼75 ng/L and ∼3 μg/L, respectively. Mean (SD range) values in saliva samples from healthy donors (n = 60) were 9.5 (1.2–21) μg/L for hBD-2 and 326 (50–931) μg/L for hBD-3. We did not detect hBD-3 in suction blister fluid (BF; n = 10) or bronchoalveolar lavage (BAL; n = 5) from healthy participants. We detected low hBD-2 peptide concentrations in BF and BAL, 0.16 (0.03–0.32) and 0.04 (0–0.049) μg/g total protein, respectively. We observed no correlation of hBD-2 in BF and saliva or BAL and saliva from the same person. In vaginal swabs from healthy women (n = 2), mean hBD-2 and -3 concentrations were 3.42 and 103 μg/g total protein, respectively. Cervicovaginal lavage from the same women contained mean concentrations of 1.46 and 55.5 μg/g total protein.Conclusion: These ELISA assays can measure inducible hBD peptide concentrations in body fluids by overcoming masking effects of anionic molecules. This approach may therefore be applicable for quantifying these peptides in health and disease.

scholarly journals Method for the determination of immune complexes by reaction with polyethylene glycol

1982 ◽  
Vol 63 (2) ◽  
pp. 10-13
Author(s):  
B. A. Molotilov ◽  
A. N. Mayansky ◽  
N. D. Pozdnyak ◽  
L. Ch. Samerkhanova
Keyword(s):  
Polyethylene Glycol ◽  
Lupus Erythematosus ◽  
Immune Complexes ◽  
Clinical Laboratory ◽  
Circulating Immune Complexes ◽  
Healthy Donors ◽  
Highly Sensitive ◽  
Photoelectric Colorimeter ◽  
Health And Disease

A study of circulating immune complexes was carried out using a reaction with polyethylene glycol. The method turned out to be simple, highly sensitive and affordable for any clinical laboratory with a photoelectric colorimeter. Analysis of the survey data of 115 healthy donors, 63 patients with rheumatoid arthritis and 16 patients with systemic lupus erythematosus made it possible to establish the level of circulating immune complexes in health and disease. The circulating immune complexes were studied in patients with rheumatism and chronic tonsillitis. To assess the results of the reaction, human aggregated gamma globulin (manufactured by Kazan NIIEM) was used.

scholarly journals High Purity Isolation of Low Density Neutrophils Casts Doubt on Their Exceptionality in Health and Disease

2021 ◽  
Vol 12 ◽  
Author(s):  
Gareth R. Hardisty ◽  
Frances Llanwarne ◽  
Danielle Minns ◽  
Jonathan L. Gillan ◽  
Donald J. Davidson ◽  
...  
Keyword(s):  
High Purity ◽  
Neutrophil Extracellular Trap ◽  
Normal Density ◽  
Low Density ◽  
Inflammatory Conditions ◽  
Functional Differences ◽  
Healthy Donors ◽  
Health And Disease

Low density neutrophils (LDNs) are described in a number of inflammatory conditions, cancers and infections and associated with immunopathology, and a mechanistic role in disease. The role of LDNs at homeostasis in healthy individuals has not been investigated. We have developed an isolation protocol that generates high purity LDNs from healthy donors. Healthy LDNs were identical to healthy normal density neutrophils (NDNs), aside from reduced neutrophil extracellular trap formation. CD66b, CD16, CD15, CD10, CD54, CD62L, CXCR2, CD47 and CD11b were expressed at equivalent levels in healthy LDNs and NDNs and underwent apoptosis and ROS production interchangeably. Healthy LDNs had no differential effect on CD4+ or CD8+ T cell proliferation or IFNγ production compared with NDNs. LDNs were generated from healthy NDNs in vitro by activation with TNF, LPS or fMLF, suggesting a mechanism of LDN generation in disease however, we show neutrophilia in people with Cystic Fibrosis (CF) was not due to increased LDNs. LDNs are present in the neutrophil pool at homeostasis and have limited functional differences to NDNs. We conclude that increased LDN numbers in disease reflect the specific pathology or inflammatory environment and that neutrophil density alone is inadequate to classify discrete functional populations of neutrophils.

scholarly journals Intraplatelet serotonin and plasma 5-hydroxyindoles in health and disease

Blood ◽  
1981 ◽  
Vol 57 (3) ◽  
pp. 505-509 ◽  
Author(s):  
RD Shuttleworth ◽  
JR O'Brien
Keyword(s):  
Myocardial Infarction ◽  
Acute Myocardial Infarction ◽  
Acetic Acid ◽  
Chest Pain ◽  
Platelet Lysate ◽  
The Other ◽  
Positive Material ◽  
Healthy Donors ◽  
Health And Disease

Abstract Using a fluorometric test sensitive to serotonin (5-HT) and to other 5- hydroxyindoles (5-HIs) it was shown that platelets take up 5-HT and that the added 5-HT and the inherent test-positive material in platelet lysate can be directly measured. However, platelets do not take up 5- hydroxyindole acetic acid or the 5-HIs from the plasma. Thus, 5-HT and the other 5-HIs can be distinguished. Various methods of liberating intraplatelets 5-HT were investigated. Several anticoagulants, temperatures, and speeds of centrifugation were employed, with no effect on the results. It was found that in healthy donors, with increasing age, there was a decrease in intraplatelet 5-HT and an increase in plasma 5-HIs. The quantities of these substances were inversely related. In acute myocardial infarction, the 5-HT was normal taking age into account, but the plasma 5-HIs were increased. In postoperative patients, the platelet 5-HT was low and the plasma 5-HIs were normal. In patients with chest pain but no myocardial infarction, both platelet 5-HT and plasma 5-HIs were normal. The relevance of these findings to 5-HT metabolism and the evidence for in vivo activation of platelets is discussed.

Sample viscosity can be a source of analytical error when discrete sampler-dilutors are used.

1981 ◽  
Vol 27 (11) ◽  
pp. 1896-1898 ◽  
Author(s):  
K M Chan ◽  
J H Ladenson
Keyword(s):  
Total Protein ◽  
Protein Concentration ◽  
Body Fluids ◽  
Sample Temperature ◽  
Total Protein Concentration ◽  
Sampling System ◽  
Hyperviscosity Syndrome ◽  
Analytical Error

Abstract Total protein concentration in the serum of a patient with hyperviscosity syndrome differed as measured by the biuret procedure in the DuPont aca (80 g/L) and the SMA 12/60 (105 g/L), owing to viscosity-dependent errors with the aca sampling system; the magnitude depended on sample temperature and volume of sample aspirated. This kind of error was not observed with the SMA 12/60 and was far less severe when a Micromedic sampler-dilutor was tested. It could be eliminated in the case of the aca by adding sample to test packs with a syringe rather than with the aca automated sampler-dilutor. We thus recommend use of the syringe method when unusually viscous samples (serum or other body fluids) are analyzed in the aca.

Measurement of deoxyribonuclease I activity in human tissues and body fluids by a single radial enzyme-diffusion method

1993 ◽  
Vol 39 (3) ◽  
pp. 448-452 ◽  
Author(s):  
D Nadano ◽  
T Yasuda ◽  
K Kishi
Keyword(s):  
Limit Of Detection ◽  
Body Fluids ◽  
Diffusion Method ◽  
Agarose Gel ◽  
Human Tissues ◽  
Deoxyribonuclease I ◽  
Gel Layer ◽  
Dark Circle ◽  
Healthy Donors ◽  
Measured Volume

Abstract In the single radial enzyme-diffusion (SRED) method for assay of deoxyribonuclease I, a precisely measured volume of the enzyme solution is dispensed into a circular well in an agarose gel layer in which DNA and ethidium bromide are uniformly distributed. A circular dark zone is formed as the enzyme diffuses from the well radially into the gel and digests substrate DNA. The diameter of the dark circle of hydrolyzed DNA increases in size with time and correlates linearly with the amount of enzyme applied to the well. Thus, the SRED can be used for quantitation of deoxyribonuclease I with a limit of detection of 2 x 10(-6) unit. This corresponds to 1 pg of purified urine deoxyribonuclease I. We measured the deoxyribonuclease I activity of 17 different human tissues and body fluids from healthy donors. Urine samples showed the greatest activity, 6.0 +/- 2.2 kilo-units/g protein (mean +/- SD). Serum deoxyribonuclease I activity was 4.4 +/- 1.8 units/L.

The Kidney and Body Fluids in Health and Disease

JAMA ◽  
1983 ◽  
Vol 250 (21) ◽  
pp. 2986
Author(s):  
George Dunea
Keyword(s):  
Body Fluids ◽  
Health And Disease

scholarly journals A NKp80-based identification strategy reveals that CD56neg NK cells are not completely dysfunctional in health and disease

2020 ◽  
Author(s):  
Ane Orrantia ◽  
Iñigo Terrén ◽  
Alicia Izquierdo-Lafuente ◽  
Juncal A. Alonso-Cabrera ◽  
Victor Sandá ◽  
...  
Keyword(s):  
Nk Cells ◽  
Good Alternative ◽  
Specific Marker ◽  
Effector Functions ◽  
Multiparametric Flow Cytometry ◽  
Healthy Donors ◽  
A Cell ◽  
Health And Disease ◽  
Lineage Markers ◽  
Identification Strategy

ABSTRACTNatural killer (NK) cells are usually identified by the absence of other lineage markers, due to the lack of a cell surface specific marker. CD56neg NK cells, classically identified as CD56negCD16+ are known to be expanded in some pathological conditions. However, studies on CD56neg NK cells had revealed different results regarding the phenotype and functionality of these cells. This could be due to, among others, the unstable expression of CD16, which hinders CD56neg NK cells identification. Hence, we aim to determine an alternative surface marker to CD16 to better identify CD56neg NK cells. Using multiparametric flow cytometry, we have found that NKp80 is a good alternative to CD16 not only in healthy donors but also in HIV-1 infected subjects and multiple myeloma patients. Furthermore, we found differences between the functionality of CD56negNKp80+ and CD56negCD16+ NK cells both in healthy donors and patients, suggesting that the effector functions of CD56neg NK cells are not as diminished as previously thought. We proposed NKp80 as a noteworthy marker to identify and accurately re-characterize human CD56neg NK cells.

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