Enumeration and phenotypic assessment of human plasmacytoid and myeloid dendritic cells in whole blood

Chowdhury, Ferdousi; Johnson, Peter; Williams, Anthony P.

doi:10.1002/cyto.a.20872

Cited by 15 publications

(12 citation statements)

References 47 publications

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“…The subsets of circulating DC precursors include precursor myeloid DCs and precursor plasmacytoid DCs (212223). These cells express complementary sets of TLRs and respond to different pathogen-associated molecular patterns by upregulating CCR7 expression and producing distinct combinations of cytokines.…”

Section: Sources and Functions Of Dcsmentioning

confidence: 99%

Dendritic Cell Dysfunction in Patients with End-stage Renal Disease

Kim

et al. 2017

Immune Netw

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End-stage renal disease (ESRD) with immune disorder involves complex interactions between the innate and adaptive immune responses. ESRD is associated with various alterations in immune function such as a reduction in polymorphonuclear leukocyte bactericidal activity, a suppression of lymphocyte proliferative response to stimuli, and a malfunction of cell-mediated immunity at the molecular level. ESRD also increases patients' propensity for infections and malignancies as well as causing a diminished response to vaccination. Several factors influence the immunodeficiency in patients with ESRD, including uremic toxins, malnutrition, chronic inflammation, and the therapeutic dialysis modality. The alteration of T-cell function in ESRD has been considered to be a major factor underlying the impaired adaptive cellular immunity in these patients. However, cumulative evidence has suggested that the immune defect in ESRD can be caused by an Ag-presenting dendritic cell (DC) dysfunction in addition to a T-cell defect. It has been reported that ESRD has a deleterious effect on DCs both in terms of their number and function, although the precise mechanism by which DC function becomes altered in these patients is unclear. In this review, we discuss the effects of ESRD on the number and function of DCs and propose a possible molecular mechanism for DC dysfunction. We also address therapeutic approaches to improve immune function by optimally activating DCs in patients with ESRD.

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Section: Sources and Functions Of Dcsmentioning

confidence: 99%

Dendritic Cell Dysfunction in Patients with End-stage Renal Disease

Kim

et al. 2017

Immune Netw

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“…Follow-up samples were also taken at day 49 after first infusion. These whole blood samples were stained to assess changes in T-, B-, and NK-cell numbers using the BD Multitest CD3 FITC/CD16 þ CD56 PE/CD45 PerCP/CD19 APC Reagent (Becton Dickinson), and DC number and activation as described previously (8). Samples were acquired using FACSCanto flow cytometers (Becton Dickinson).…”

Section: Pharmacodynamic Assessmentsmentioning

confidence: 99%

Clinical and Biological Effects of an Agonist Anti-CD40 Antibody: A Cancer Research UK Phase I Study

Johnson

Challis

Chowdhury

et al. 2015

Clinical Cancer Research

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Purpose: This phase I study aimed to establish the biologic effects and MTD of the agonistic IgG1 chimeric anti-CD40 antibody ChiLob7/4 in patients (pts) with a range of CD40-expressing solid tumors and diffuse large B-cell lymphoma, resistant to conventional therapy. Potential mechanisms of action for agonistic anti-CD40 include direct cytotoxic effects on tumor cells and conditioning of antigen-presenting cells.Experimental Design: ChiLob7/4 was given by IV infusion weekly for 4 doses at a range from 0.5 to 240 mg/dose. Validated ELISAs were used to quantify ChiLob7/4 in serum and test for anti-chimeric MAb (HACA) responses. Pharmacodynamic assessments included quantitation of T-cell, natural killer-cell, and B-cell numbers and activation in blood by flow cytometry and a panel of cytokines in plasma by Luminex technology. Planned dose escalation was in cohorts of 3 patients until MTD or biologic effect, defined as reduction of peripheral blood CD19 þ B cells to 10% or less of baseline.Results: Twenty-nine courses of treatment were given to 28 subjects. The MTD was 200 mg Â 4, with dose-limiting toxicity of liver transaminase elevations at 240 mg. At 200 mg (range between 2.1 mg/kg and 3.3 mg/kg based on patient body weight), the trough level pretreatment was above 25 mg/mL. Grade 1-2 infusion reactions were seen above the dose of 16 mg, but could be prevented with single-dose corticosteroid premedication. HACA responses were seen after doses between 1.6 mg and 50 mg, but not above this. There were dose-dependent falls in blood Bcell numbers accompanied by reduced expression of CD21, and transient reductions in NK cell numbers with increased CD54 expression from 50 mg upward. MIP-1b and IL12 plasma concentrations rose after doses above 16 mg. Fifteen of 29 treatments were accompanied by disease stabilization for a median 6 months, the longest for 37 months.Conclusions: ChiLob7/4 can activate B and NK cells at doses that can be administered safely, and should be tested in combination with other antibodies and chemotherapy agents.

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“…Samples were acquired using the BD FACS Canto II and analyzed using the FACS Diva software. The gating strategy used was as reported (17). Briefly, leukocytes were gated using a side scatter versus CD45 gate.…”

Section: Dendritic Cell Flow Cytometric Analysismentioning

confidence: 99%

Ex Vivo Assays of Dendritic Cell Activation and Cytokine Profiles as Predictors of In Vivo Effects in an Anti-Human CD40 Monoclonal Antibody ChiLob 7/4 Phase I Trial

Chowdhury

Johnson

Glennie

et al. 2014

Cancer Immunology Research

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Immunostimulatory antibodies entering the clinic create challenge in terms of not only pharmacodynamics for monitoring anticipated mechanisms but also predetermining cytotoxicity. We show the use of ex vivo wholeblood samples to predict the activation requirements, cytokine signature, and adverse events of an anti-human-CD40 chimeric IgG1 antibody, ChiLob 7/4. Assessments were initially undertaken on human myeloid (mDC1) and plasmacytoid (pDC) dendritic cells, in which an absolute need for cross-linking was shown through the upregulation of activation markers CD83 and CCR7. Subsequent cytokine secretion evaluations of ex vivo whole blood showed the cross-linked antibody-induced increases in MIP1b, interleukin (IL)-8, IL-12, TNFa, and IL-6. This cytokine signature compared favorably with the Toll-like receptor (TLR) ligand lipopolysaccharide (LPS), in which levels of TNFa and IL-6 were significantly higher, suggesting a less intense proinflammatory response and possible modified cytokine release syndrome when used in human trials. Following first-in-human use of this agent within a dose escalation study, in vivo evaluations of dendritic cell activation and secreted cytokines closely matched the predetermined immunomonitoring endpoints. Patients showed a comparable pattern of MIP1b, IL-8, and IL-12 secretion, but no TNFa and IL-6 were identified. Mild symptoms relating to a cytokine release syndrome were seen at an equivalent dosage to that observed for dendritic cell activation and cytokine release. In summary, ChiLob 7/4 induces a distinctive pattern of dendritic cell activation and cytokine secretion in ex vivo assays that can be predictive of in vivo responses. Such preclinical approaches to monoclonal antibody evaluation may inform both the starting dosages and the anticipated cytokine release events that could occur, providing a valuable adjunct for future first-in-human assessments of immunostimulatory antibodies. Cancer Immunol Res; 2(3); 229-40. Ó2013 AACR.

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Enumeration and phenotypic assessment of human plasmacytoid and myeloid dendritic cells in whole blood

Cited by 15 publications

References 47 publications

Dendritic Cell Dysfunction in Patients with End-stage Renal Disease

Dendritic Cell Dysfunction in Patients with End-stage Renal Disease

Clinical and Biological Effects of an Agonist Anti-CD40 Antibody: A Cancer Research UK Phase I Study

Ex Vivo Assays of Dendritic Cell Activation and Cytokine Profiles as Predictors of In Vivo Effects in an Anti-Human CD40 Monoclonal Antibody ChiLob 7/4 Phase I Trial

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