scholarly journals Methods for Obtaining Purified Lymphocytes, Glass-adherent Mononuclear Cells, and a Population Containing Both Cell Types From Human Peripheral Blood

Blood ◽  
1972 ◽  
Vol 40 (1) ◽  
pp. 77-89 ◽  
Author(s):  
William R. Levis ◽  
Jay H. Robbins
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Human Leukocyte ◽  
Culture Fluid ◽  
Cell Types ◽  
Peripheral Blood Lymphocytes ◽  
Limiting Factor ◽  
Lymphocyte Populations ◽  
Two Populations

Abstract Methods are presented for obtaining simultaneously or separately two populations of cells from human peripheral blood, lymphocytes and monocytes, both of which are required to obtain blastogenesis and DNA synthesis in human leukocyte cultures. A simple 5-min centrifugation of heparinized blood yields a mononuclear leukocyte culture fluid containing 70-90% lymphocytes with few granulocytes but with sufficient numbers of monocytes so that the latter cell is not a limiting factor in the blastogenic reaction. A method is also presented for removing both granulocytes and monocytes from lymphocyte populations in a manner that permits monitoring and choice of the degree of lymphocyte purification. A method is also presented for obtaining glass-adherent mononuclear cells that do not undergo blastogenesis but will enable suitably stimulated "purified" lymphocytes to undergo blastogenesis. Studies of the function and morphology of these different cell populations are presented.

scholarly journals Characteristics of a Pathogenic Molecular Clone of an End-Stage Serum-Derived Variant of Simian Immunodeficiency Virus (SIVF359)

2001 ◽  
Vol 75 (19) ◽  
pp. 9328-9338 ◽  
Author(s):  
Lennart Holterman ◽  
Rob Dubbes ◽  
James Mullins ◽  
Gerald Learn ◽  
Henk Niphuis ◽  
...  
Keyword(s):  
Simian Immunodeficiency Virus ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Peripheral Blood Mononuclear ◽  
Molecular Clone ◽  
Immunodeficiency Virus ◽  
End Stage

ABSTRACT End-stage simian immunodeficiency virus (SIV) isolates are suggested to be the most fit of the evolved virulent variants that precipitate the progression to AIDS. To determine if there were common characteristics of end-stage variants which emerge from accelerated cases of AIDS, a molecular clone was derived directly from serum following in vivo selection of a highly virulent SIV isolate obtained by serial end-stage passage in rhesus monkeys (Macaca mulatta). This dominant variant caused a marked cytopathic effect and replicated to very high levels in activated but not resting peripheral blood lymphocytes. Furthermore, although this clone infected but did not replicate to detectable levels in rhesus monocyte-derived macrophages, these cells were able to transmit infection to autologous T cells upon contact. Interestingly, although at low doses this end-stage variant did not use any of the known coreceptors except CCR5, it was able to infect and replicate in human peripheral blood mononuclear cells homozygous for the Δ32 deletion of CCR5, suggesting the use of a novel coreceptor. It represents the first pathogenic molecular clone of SIV derived from viral RNA in serum and provides evidence that not only the genetic but also the biological characteristics acquired by highly fit late-stage disease variants may be distinct in different hosts.

scholarly journals Spurious detection of terminal deoxynucleotidyl transferase in phytohemagglutinin-stimulated lymphocytes

Blood ◽  
1986 ◽  
Vol 68 (1) ◽  
pp. 310-312
Author(s):  
JA Fletcher ◽  
R Bell ◽  
M Koekebakker ◽  
AA Dowers ◽  
RP McCaffrey ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Protein S ◽  
Peripheral Blood Lymphocytes ◽  
Lymphoid Cells ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Peripheral Blood Mononuclear ◽  
Biochemical Assays ◽  
Inducible Protein

Expression of terminal transferase (TdT) is believed to be restricted to primitive lymphoid cells; recently, however, indirect immunofluorescent (IF) assays have been used to demonstrate the apparent presence of TdT in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocytes and in various nonlymphoid malignancies. Using an IF assay, we found that a heteroantiserum to TdT reacted with cultured and PHA-stimulated human peripheral blood mononuclear cells, but we were unable to confirm the presence of TdT in these cells using immunoblotting and biochemical assays. We conclude that the IF results are spurious and most likely represent recognition by the heteroantiserum of inducible protein(s) other than TdT.

scholarly journals Effects of 2-amino-4,6-di-tert-butylphenol derivatives on the viability and functional state of human peripheral blood lymphocytes

2020 ◽  
pp. 19-28
Author(s):  
Darya B. Nizheharodava ◽  
Galina A. Ksendzova ◽  
Aliaksei G. Sysa ◽  
Mariya Yu. Yurkevich ◽  
Maryna V. Labai ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Immunomodulatory Activity ◽  
Peripheral Blood Mononuclear ◽  
Blood Lymphocytes ◽  
Tert Butyl ◽  
Human Peripheral Blood Lymphocytes

Derivatives of 2-amino-4,6-di-tert-butylphenol exhibit antiviral properties and radical regulatory activity against various types of organic radicals which determines the actuality of their further investigation. But the question of aminophenol derivatives immunomodulatory activity remains open. In this regard, the aim of the study was to assess the effects of 2-amino-4,6-di-tert-butylphenol derivatives on the viability and functional potential of human peripheral blood lymphocytes. As a result of the studies, it was shown that aminophenol compounds at concentrations of 10–5–10–7 mol did not exert a toxic effect while at a concentration of 10–4 mol showed a cytotoxic effect due to the induction of secondary necrosis. Compounds N-(2-hydroxy-3,5-di-tert-butylphenyl)-4-methylbenzenesulfonamide and 2,4-di-tert-butyl-6-morpholinophenol at a concentration of 10–6 mol stimulated the extracellular production of α-interferon by peripheral blood mononuclear cells and intracellular production of γ-interferon by CD3+T-lymphocytes. An immunosuppressive effect (more than 50 %) of N-(2-hydroxy-3,5-di-tert-butylphenyl)-4-methylbenzenesulfonamide and 2,4-di-tert-butyl-6-morpholinophenol compounds at a concentration of 10–5 mol was revealed to the mitogen-induced proliferation of T-lymphocytes.

scholarly journals MicroRNA levels quantified in whole blood varies from PBMCs

F1000Research ◽  
2014 ◽  
Vol 3 ◽  
pp. 183 ◽  
Author(s):  
Sadaf Atarod ◽  
Hannah Smith ◽  
Anne Dickinson ◽  
Xiao-Nong Wang
Keyword(s):  
Whole Blood ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Microrna Expression ◽  
Peripheral Blood Mononuclear ◽  
Total Rna ◽  
Disease Stages ◽  
Different Cell Types

MicroRNAs are non-coding RNAs that negatively regulate mRNA expression and play significant roles in both health and disease. Differential microRNA expression has been used to aid diagnosis and discriminate disease stages. The accuracy and reliability of microRNA expression measurement is of utmost importance. Quantification of microRNA expression in human peripheral blood is commonly detected using total RNA extracted via different methods. To date, no convincing data are available showing whether microRNA quantification results can be influenced by the use of total RNA extracted from whole blood or peripheral blood mononuclear cells (PBMCs). This study examined miR-146a-5p and miR-155-5p expression using total RNA extracted in parallel from whole blood and PBMCs of 14 healthy volunteers. The data showed that the quantification of miRNA using total RNA extracted from whole blood varied from that of PBMCs, indicating that the miRNA expression was a result of all the different cell-types present in whole blood. Our results suggested that the source of total RNA and the statistical analyses performed are crucial considerations when designing miRNA research.

scholarly journals MicroRNA levels quantified in whole blood varies from PBMCs

F1000Research ◽  
2015 ◽  
Vol 3 ◽  
pp. 183 ◽  
Author(s):  
Sadaf Atarod ◽  
Hannah Smith ◽  
Anne Dickinson ◽  
Xiao-Nong Wang
Keyword(s):  
Whole Blood ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Microrna Expression ◽  
Peripheral Blood Mononuclear ◽  
Total Rna ◽  
Disease Stages ◽  
Different Cell Types

MicroRNAs are non-coding RNAs that negatively regulate mRNA expression and play significant roles in both health and disease. Differential microRNA expression has been used to aid diagnosis and discriminate disease stages. The accuracy and reliability of microRNA expression measurement is of utmost importance. Quantification of microRNA expression in human peripheral blood is commonly detected using total RNA extracted via different methods. To date, no convincing data are available showing whether microRNA quantification results can be influenced by the use of total RNA extracted from whole blood or peripheral blood mononuclear cells (PBMCs). This study examined miR-146a-5p and miR-155-5p expression using total RNA extracted in parallel from whole blood and PBMCs of 14 healthy volunteers. The data showed that the quantification of miRNA using total RNA extracted from whole blood varied from that of PBMCs, indicating that the miRNA expression was a result of all the different cell-types present in whole blood. Our results suggested that the source of total RNA and the statistical analyses performed are crucial considerations when designing miRNA research.

scholarly journals Large granular lymphocytes in human peripheral blood: ultrastructural and cytochemical characterization of the granules

Blood ◽  
1982 ◽  
Vol 59 (2) ◽  
pp. 277-283 ◽  
Author(s):  
CE Grossi ◽  
A Cadoni ◽  
A Zicca ◽  
A Leprini ◽  
M Ferrarini
Keyword(s):  
Golgi Apparatus ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Single Cells ◽  
Cell Types ◽  
Large Granular Lymphocytes ◽  
Enzyme Cytochemistry ◽  
Acetate Esterase ◽  
Granular Lymphocytes

Abstract Large granular lymphocytes (LGL) are defined as nonadherent mononuclear cells with cytoplasmic azurophilic granules, avid receptors for the Fc portion of IgG, and cytotoxic functions (NK or ADCC activities). In the present study, the granules of LGL isolated from human peripheral blood have been analyzed by enzyme cytochemistry and electron microscopy. It had been found that: (1) in the single cells, granules at different stages of maturation could be detected: in addition, packaging of the granules took place in the proximity of the Golgi apparatus, which is similar to that seen in secretory cell types. (2) Acid phosphatase (AP) was observed within the granules and the vesicles located in the Golgi area: the Golgi apparatus identified through its thiamine pyrophosphatase-positivity was consistently negative for AP. (3) Alpha naphthyl-acetate esterase (ANAE) activity was localized in the granules as well as on the membrane of LGL and monocytes. (4) The ANAE activity of LGL was of the monocytic and not of the lymphocytic type, as shown by NaF inhibition. (5) The LGL granules, although identifiable as primary lysosomes, were not involved in the process of phagocytosis, since LGL failed consistently to ingest latex particles or opsonized red cells.

scholarly journals Evasion of Host Defenses by Measles Virus: Wild-Type Measles Virus Infection Interferes with Induction of Alpha/Beta Interferon Production

2000 ◽  
Vol 74 (16) ◽  
pp. 7478-7484 ◽  
Author(s):  
Denise Naniche ◽  
Annie Yeh ◽  
Danelle Eto ◽  
Marianne Manchester ◽  
Robert M. Friedman ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Measles Virus ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Antiviral Immunity ◽  
Peripheral Blood Lymphocytes ◽  
Wild Type ◽  
Peripheral Blood Mononuclear ◽  
Double Stranded Rna ◽  
Alpha Beta

ABSTRACT Measles is a highly contagious disease currently responsible for over one million childhood deaths, particularly in the developing world. Since alpha/beta interferons (IFNs) are pivotal players both in nonspecific antiviral immunity and in specific cellular responses, their induction or suppression by measles virus (MV) could influence the outcome of a viral infection. In this study we compare the IFN induction and sensitivity of laboratory-passaged attenuated MV strains Edmonston and Moraten with those of recent wild-type viruses isolated and passaged solely on human peripheral blood mononuclear cells (PBMC) or on the B958 marmoset B-cell line. We report that two PBMC-grown wild-type measles isolates and two B958-grown strains of MV induce 10- to 80-fold-lower production of IFN by phytohemagglutinin-stimulated peripheral blood lymphocytes (PBL) compared to Edmonston and Moraten strains of measles. Preinfection of PBL with these non-IFN-inducing MV isolates prevents Edmonston-induced but not double-stranded-RNA-induced IFN production. This suggests that the wild-type viruses can actively inhibit Edmonston-induced IFN synthesis and that this is not occurring by double-stranded RNA. Furthermore, the wild-type MV is more sensitive than Edmonston MV to the effect of IFN. MV is thus able to suppress the synthesis of the earliest mediator of antiviral immunity, IFN-α/β. This could have important implications in the virulence and spread of MV.

scholarly journals Large granular lymphocytes in human peripheral blood: ultrastructural and cytochemical characterization of the granules

Blood ◽  
1982 ◽  
Vol 59 (2) ◽  
pp. 277-283 ◽  
Author(s):  
CE Grossi ◽  
A Cadoni ◽  
A Zicca ◽  
A Leprini ◽  
M Ferrarini
Keyword(s):  
Golgi Apparatus ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Single Cells ◽  
Cell Types ◽  
Large Granular Lymphocytes ◽  
Enzyme Cytochemistry ◽  
Acetate Esterase ◽  
Granular Lymphocytes

Large granular lymphocytes (LGL) are defined as nonadherent mononuclear cells with cytoplasmic azurophilic granules, avid receptors for the Fc portion of IgG, and cytotoxic functions (NK or ADCC activities). In the present study, the granules of LGL isolated from human peripheral blood have been analyzed by enzyme cytochemistry and electron microscopy. It had been found that: (1) in the single cells, granules at different stages of maturation could be detected: in addition, packaging of the granules took place in the proximity of the Golgi apparatus, which is similar to that seen in secretory cell types. (2) Acid phosphatase (AP) was observed within the granules and the vesicles located in the Golgi area: the Golgi apparatus identified through its thiamine pyrophosphatase-positivity was consistently negative for AP. (3) Alpha naphthyl-acetate esterase (ANAE) activity was localized in the granules as well as on the membrane of LGL and monocytes. (4) The ANAE activity of LGL was of the monocytic and not of the lymphocytic type, as shown by NaF inhibition. (5) The LGL granules, although identifiable as primary lysosomes, were not involved in the process of phagocytosis, since LGL failed consistently to ingest latex particles or opsonized red cells.

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