Immuno-monitoring of CD8+ T cells in whole blood versus PBMC samples

Appay, Victor; Reynard, Séverine; Voelter, Verena; Romero, Pedro; Speiser, Daniel E.; Leyvraz, Serge

doi:10.1016/j.jim.2005.11.007

Cited by 48 publications

(37 citation statements)

References 14 publications

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“…greatly facilitates clinical investigation of MDSC. Analysis of samples which have undergone manipulations yields mostly qualitative information [4]. Additional purification steps may alter the natural frequency of MDSC in circulation and may unintentionally introduce biases.…”

Section: Discussionmentioning

confidence: 99%

Increased circulating myeloid-derived suppressor cells correlate with clinical cancer stage, metastatic tumor burden, and doxorubicin–cyclophosphamide chemotherapy

Díaz-Montero

Salem

Nishimura

et al. 2008

Cancer Immunol Immunother

1,096

889

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Abnormal accumulation of myeloid-derived suppressor cells (MDSC) is an important mechanism of tumor immune evasion. Cyclophosphamide (CTX) has also been shown in non-tumor bearing animals to cause transient surges in MDSC. Knowledge of MDSC is primarily based on preclinical work, and to date only few published studies have involved cancer patients. The goal of this study was to test the hypothesis that circulating MDSC levels correlate with clinical cancer stage, CTX-based chemotherapy, and metastatic tumor burden. Whole blood was collected from 106 newly diagnosed solid tumor patients (stages I-IV). Percentages of circulating MDSC (Lin −/Lo , HLA DR−, CD33 + CD11b + ) were determined prior to initiation of systemic therapy. In 17 early stage breast cancer patients receiving doxorubicin-cyclophosphamide chemotherapy every 14 days (ddAC) blood was collected on day 1 of each cycle. Circulating MDSC were significantly increased in cancer patients of all stages relative to healthy volunteers. A significant correlation between circulating MDSC and clinical cancer stage was also observed. Moreover, among stage IV patients, those with extensive metastatic tumor burden had the highest percent and absolute number of MDSC. Significant increases in circulating MDSC were observed with ddAC NIH Public AccessAuthor Manuscript Cancer Immunol Immunother. Author manuscript; available in PMC 2012 July 23.

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Section: Discussionmentioning

confidence: 99%

Increased circulating myeloid-derived suppressor cells correlate with clinical cancer stage, metastatic tumor burden, and doxorubicin–cyclophosphamide chemotherapy

Díaz-Montero

Salem

Nishimura

et al. 2008

Cancer Immunol Immunother

1,096

889

View full text Add to dashboard Cite

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“…The protocol for whole blood staining was described previously. 15,16 To assess absolute cell count, 50 L of whole blood was added and stained in Trucount tubes according to the manufacturer's directions (BD Biosciences). Data were acquired on a BD FACSCalibur flow cytometer (BD Biosciences) and analyzed with Cell Quest Version 5.2.1 and Multiset Version 2.2.1 (BD Biosciences) software.…”

Section: Immunophenotyping Of Peripheral Bloodmentioning

confidence: 99%

Immunosuppressive CD14+HLA-DRlow/− monocytes in B-cell non-Hodgkin lymphoma

Lin

Gustafson

Bulur

et al. 2011

Blood

224

225

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Immunosuppression is a known risk factor for B-cell non-Hodgkin lymphoma (NHL), yet mechanisms of tumor-associated immunosuppression remain to be fully characterized. We examined the immunophenotype of 40 NHL patients and 27 age-matched healthy volunteers to better understand systemic immune suppression. NHL peripheral blood mononuclear cells had significantly decreased interferon-␥ production and proliferation. This suppression was not the result of regulatory T cells, interleukin-6 or interleukin-10, as these factors were not different between NHL and healthy volunteers (controls). We were able to restore T-cell proliferation by removing NHL monocytes, suggesting that these monocytes are suppressive. This suppression was mediated in part through arginine metabolism as exogenous arginine supplementation partially overcame monocytes' suppression of T-cell proliferation in vitro and NHL patients had elevated arginase I in their plasma. NHL monocytes had impaired STAT1 phosphorylation and interferon-␣ production to CpG stimulation and a dendritic cell differentia- IntroductionSystemic immune suppression is often seen in cancer patients and is thought to contribute to patient morbidity via tumor-mediated immune evasion. Indeed, patients with compromised immune systems, such as those with HIV infection, or on immunosuppressive medications are at increased risk of developing non-Hodgkin lymphoma (NHL). 1,2 Polymorphisms in host germline immune genes also have been associated with risk of developing NHL 3 as well as survival in NHL patients. 4 Conversely, a small percentage of patients with indolent NHL have spontaneous regression of their disease without any treatment, 5 possibly linked to a host antitumor immune response. The presence of host immune cells in the tumor microenvironment has also been correlated with treatment outcome and survival. [6][7][8][9] Of note, reduction in absolute count of circulating lymphocytes has been identified as a poor prognostic factor for overall survival in newly diagnosed NHL 10,11 as well as a predictor of poor treatment response. 9,12,13 Although evidence for the role of immune suppression in tumor establishment and pathogenesis is unquestionable, the mechanisms and the cellular phenotype of systemic immune suppression in NHL patients remain to be fully characterized.In this study, we investigated the qualitative and quantitative systemic immune suppression in B-cell NHL. We found circulating mononuclear cells had suppressed interferon-␥ (IFN-␥) recall response and proliferative capacity. These suppressed functions were mediated by circulating monocytes and partly mediated via arginine metabolism. These monocytes also had other impaired adaptive immune response including a decreased capacity to generate mature dendritic cells. In addition, the innate immune response of these monocytes to CpG stimulation was impaired as measured by intracellular STAT1 phosphorylation and interferonalpha (IFN-␣) production. These multifactorial suppressive functions of monocytes in NHL were corre...

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“…The screening assay described in this paper is distinct from the previous attempts in that it measures the response of dilute whole blood to mAbs immobilized on Protein A beads. Development of an assay that uses whole blood may be important because the isopycnic gradient process used to isolate PBMC can change the composition and ratio of cells in the culture [33,34]. We feel the use of whole blood more closely mimics the in vivo environment, and therefore, should provide results that more closely resemble what will happen when mAbs are given to patients and thus may have superior predictive value.…”

Section: Discussionmentioning

confidence: 99%

“…This is thought to be due to a well-ordered molecular orientation favoring steric accessibility of the antigen to the antibody [45]. The drawback to using a bacterial protein to immobilize mAbs is that the bacterial antigen itself may cause release of cytokines from monocytes and lymphocytes in vitro [33,46]. In addition there are slight differences in the amount of human or mouse IgG Protein A binds [47,48].…”

Section: Discussionmentioning

confidence: 99%