Whole‐blood leukoreduction filters are a source for cryopreserved cells for phenotypic and functional investigations on peripheral blood lymphocytes

Néron, Sonia; Dussault, Nathalie; Racine, Claudia

doi:10.1111/j.1537-2995.2006.00772.x

Cited by 27 publications

(32 citation statements)

References 25 publications

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“…Despite the limited number of severe cases, given the relevance of these B-cell differentiation markers we determined the frequencies of CD19-, CD27-, CD38-, and CD138-positive cells within the PBMCs of 10 SLE patient samples by flow cytometry ( Table 2). The frequencies of CD19 + cells and CD27 + B cells, ranging respectively from 2-15% and 15-48%, were not significantly different from the values obtained with HC cells and similar to those previously reported for healthy individuals (Néron et al 2006). In contrast, the frequency of CD138 + cells, ranging from 1.4-10%, was well above normal ranges (p<0.002) compared with HC (0.5±0.4%), which were close to the expected normal frequency of 0.5% (Harada et al 1996).…”

Section: Cd138 + Cells Are Present At High Frequencies In Mononuclearsupporting

confidence: 90%

“…A maximum of 14 tubes from each patient were collected, pooled, and diluted in one volume of 10 mM phosphate-buffered saline, pH 7.4. Leukocytes from healthy individuals were recovered from leukoreduction systems, as described previously (Néron et al 2006;Néron et al 2007), after informed consent and following approval by the Research Ethics Committee of Héma-Québec. Peripheral blood mononuclear cells (PBMCs) were prepared from patients and controls by density centrifugation over Ficoll-Paque (GE Healthcare Bio--Sciences Inc., Baie d'Urfé, QC, Canada) (Fecteau and Néron 2003) and stored frozen until B-cell purification.…”

Section: Blood Sample Collection From Sle Patients and Healthy Indivimentioning

confidence: 99%

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CD40-activated B cells from patients with systemic lupus erythematosus can be modulated by therapeutic immunoglobulins in vitro

Néron

Boire

Dussault

et al. 2009

Arch. Immunol. Ther. Exp.

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Introduction: Aberrant signaling within and between B and T cells, considered to be central in systemic lupus erythematosus (SLE), could depend on enhanced CD40-CD154 activation. As a result, autoreactive B cells, normally anergic, differentiate and secrete antibodies attacking several normal tissues. Thus restorating B cell homeostasis might help control this disease. In this study, two facets of SLE B cells were investigated, namely their in vitro response to CD40-CD154 and the effect of treatment with human immunoglobulins for intravenous use (IVIg). Materials and Methods: Blood samples from SLE patients and healthy volunteers were obtained and used to isolate B cells, which were activated through CD40 in the presence or absence of IVIg. The phenotype, proliferation, and differentiation of the SLE B cells were determined and compared with those of control B cells using flow cytometry and standard ELISA. Results: In this model, CD40-activated SLE B cells, as control B cells, proliferated and differentiated and were characterized by the emergence of CD19 lo CD38 ++ CD138 + CD27 ++ cells. IVIg treatment of the CD40-activated SLE B cells resulted in higher differentiation, characterized by increased secretion rates of IgG and IgM, as reported previously for control B cells. Conclusions: Taken as a whole, such accelerated differentiation of CD40-activated B cells suggests that IVIg may participate in re-equilibration of the antibody repertoire by replacing pathological antibodies by de novo harmless antibodies.

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Section: Cd138 + Cells Are Present At High Frequencies In Mononuclearsupporting

confidence: 90%

Section: Blood Sample Collection From Sle Patients and Healthy Indivimentioning

confidence: 99%

CD40-activated B cells from patients with systemic lupus erythematosus can be modulated by therapeutic immunoglobulins in vitro

Néron

Boire

Dussault

et al. 2009

Arch. Immunol. Ther. Exp.

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“…They observed that the percentage of T cells, B cells, monocytes and dendritic cells was largely similar between PBMCs isolated from LRS cones or buffy coats, but that PBMCs from LRS cones contain more granulocytes [3–5, 7]. Dendritic cells isolated from LRS cones were functional and capable of activating T cell responses [3, 6].…”

Section: Introductionmentioning

confidence: 99%

“…Dendritic cells isolated from LRS cones were functional and capable of activating T cell responses [3, 6]. B cells isolated from LRS cones have similar expression of surface markers and ability to be activated through CD40 compared to B cells isolated from buffy coats [5]. Less is known about the functionality of T cells isolated from LRS cones compared to buffy coats.…”

Section: Introductionmentioning

confidence: 99%

TCR-mediated functions are enhanced in activated peripheral blood T cells isolated from leucocyte reduction systems

Tremblay

Houtman

2015

Journal of Immunological Methods

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Buffy coats are the most common method for the acquisition of activated primary human T cells for research or clinical applications, but recently leukocyte reduction system (LRS) cones have emerged as a viable source for these cells. In this study, we determined if activated human T cells derived from buffy coats or LRS cones had different functionality. No changes in the expression of surface receptors were observed except for a significant increase in CD44 expression on T cells isolated from LRS cones. LRS cone-derived T cells trended towards higher receptor-mediated cytokine production and had significantly increased donor-to-donor variability in IFN-γ production. TCR-induced ERK1/ERK2 and AKT phosphorylation was also increased in T cells isolated from LRS cones. In conclusion, LRS cones are an excellent source of T cells for clinical and research applications, but these cells have subtle functional differences from T cells isolated using standard buffy coats.

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“…Estudos já mostraram que a frequência leucocitária presente nos filtros de leucorredução é similar aquela encontrada no sangue (51,62,66,67), e que a frequência de células mononucleares presentes da CLR é semelhante à frequência nos filtros (45 …”

Section: Discussionunclassified

Análise de populações leucocitárias em doadores de plaquetas e em câmara de leucorredução.

Borges¹

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Versão corrigida. A versão original eletrônica se encontra-se disponível tanto na Biblioteca do ICB quanto na Biblioteca Digital de Teses e Dissertações da USP (BDTD). São Paulo 2014À Deus dedico meu trabalho e minha vida. AGRADECIMENTOSAgradeço em primeiro lugar a Deus, que me guiou até aqui, que me mostrou o caminho, que me deu forças quando mais precisava e me deu sabedoria e inteligência durante todas as etapas desse projeto.Ao meu amado esposo Joel, que me apoiou em uma jornada incerta, mas que culminou na realização desse desejo tão especial. Agradeço por me incentivar a ir atrás dos meus sonhos. Por me apoiar. Por secar minhas lágrimas. Por me lembrar de que sempre há uma saída. Agradeço por abrir mão dos seus sonhos para que eu pudesse realizar os meus.Sem essa pessoa tão especial nada disso seria possível.Agradeço aos meus pais, por todo apoio e por serem a base de toda minha vida. Plateletpheresis is an automatized procedure to obtain high purity platelet for transfusions. From this procedure it's possible to obtain a byproduct: The Leukoreduction System Chamber (LRSC), which is discarded at the end of donation process. This type of donation allows 24 donation/year, but the consequences of frequent donations are poorly investigated. Therefore, we identified and quantified leukocytes of frequent and first time platelet donor. Also, was evaluated the viability, for research, of mononuclear cells recovery from LRSC. The total number of mononuclear cells was higher in LRSC than in peripheral blood samples, but the frequencies were similar in both the samples. Activation state and functional capacity (measured by cell proliferation and cytokine production) were similar in both, blood and LRSC mononuclear cells, as well as spontaneous apoptosis. Among frequent (6 or more donations in 1 year) and first time donor, there was no difference in the leukocyte total number, suggesting that frequent donation do not modify these cells.

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Whole‐blood leukoreduction filters are a source for cryopreserved cells for phenotypic and functional investigations on peripheral blood lymphocytes

Abstract: Leukoreduction filters represent a valuable source of PBMNCs. These cells could be easily recovered to prepare frozen cell banks useful in basic phenotypic and functional analyses involving the main subsets of B cells and the global T-cell population.

Cited by 27 publications

References 25 publications

CD40-activated B cells from patients with systemic lupus erythematosus can be modulated by therapeutic immunoglobulins in vitro

CD40-activated B cells from patients with systemic lupus erythematosus can be modulated by therapeutic immunoglobulins in vitro

TCR-mediated functions are enhanced in activated peripheral blood T cells isolated from leucocyte reduction systems

Análise de populações leucocitárias em doadores de plaquetas e em câmara de leucorredução.

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