Neutropenia induced in outbred mice by a simplified low-dose cyclophosphamide regimen: characterization and applicability to diverse experimental models of infectious diseases

Zuluaga, Andres F.; Salazar, Beatriz E.; Rodríguez, Carlos A.; Zapata, Ana X; Agudelo, María; Vesga, Ómar

doi:10.1186/1471-2334-6-55

Cited by 165 publications

(154 citation statements)

References 24 publications

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“…Cyclophosphamide is a common chemotherapeutic agent that is also used for treatment of autoimmune disorders. While the drug is well-known to cause neutropenia (27), it has a number of unrelated side effects as well (28)(29)(30). As such, it was unclear if the neutropenia associated with cyclophosphamide was responsible for the development of murine pneumonia.…”

Section: Discussionmentioning

confidence: 99%

Neutrophil Depletion Causes a Fatal Defect in Murine Pulmonary Staphylococcus aureus clearance

Robertson

Perrone

McConnell

et al. 2008

Journal of Surgical Research

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Section: Discussionmentioning

confidence: 99%

Neutrophil Depletion Causes a Fatal Defect in Murine Pulmonary Staphylococcus aureus clearance

Robertson

Perrone

McConnell

et al. 2008

Journal of Surgical Research

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“…Therefore Regimen 4 should be used preferentially, to study neutropenic diseases. Regimens 1 and 3 are commonly found in the literature in neutropenic infection studies (28).…”

Section: Use Of the Volumetric Assay For Anc To Follow The Kinetics Omentioning

confidence: 99%

Absolute counting of neutrophils in whole blood using flow cytometry

et al. 2014

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Absolute neutrophil count (ANC) is used clinically to monitor physiological dysfunctions such as myelosuppression or infection. In the research laboratory, ANC is a valuable measure to monitor the evolution of a wide range of disease states in disease models. Flow cytometry (FCM) is a fast, widely used approach to confidently identify thousands of cells within minutes. FCM can be optimised for absolute counting using spiked-in beads or by measuring the sample volume analysed. Here we combine the 1A8 antibody, specific for the mouse granulocyte protein Ly6G, with flow cytometric counting in straightforward FCM assays for mouse ANC, easily implementable in the research laboratory. Volumetric and Trucount TM bead assays were optimized for mouse neutrophils, and ANC values obtained with these protocols were compared to ANC measured by a dual-platform assay using the Orphee Mythic 18 veterinary haematology analyser. The single platform assays were more precise with decreased intra-assay variability compared with ANC obtained using the dual protocol. Defining ANC based on Ly6G expression produces a 15% higher estimate than the dual protocol. Allowing for this difference in ANC definition, the flow cytometry counting assays using Ly6G can be used reliably in the research laboratory to quantify mouse ANC from a small volume of blood. We demonstrate the utility of the volumetric protocol in a time-course study of chemotherapy induced neutropenia using four drug regimens. V C 2014 International Society for Advancement of CytometryKey terms absolute count; neutrophils; ANC; flow cytometry; volumetry; single-platform; doubleplatform THE concentration of blood circulating neutrophils is considerably altered by a wide range of physical and chemical stimuli such as microbial infections, gamma irradiation, and chemotherapy. Fluctuations outside of the normal range of absolute neutrophil count (ANC) correlate with increased morbidity and mortality (1). ANC is a relevant clinical parameter (2) and is routinely obtained from automated haematology instruments. However, more than 25% of samples require additional manual review of peripheral blood smears, usually because measurements fall outside the acceptable range of the instrument (3). Manual smear inspection increases workload and cost while reducing ANC accuracy. Hence, standardized flow cytometry (FCM) absolute count protocols are being pursued for implementation in the clinic (4). Even in research laboratories where flow cytometers are prevalent, ANC is obtained with veterinary haematology analysers (5) and often outsourced (6). At the time of writing, there is no published protocol to measure mouse ANC using a single, straightforward, flow cytometric assay.FCM protocols are available for absolute counting of other white blood cell (WBC) subpopulations in animal experimental models and in humans (7). Volumetric FCM involves the systematic measurement of all volumes throughout the procedure, including the volume acquired on the flow cytometer. Therefore, with a simple

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“…Mice received a total dose of 250 mg/kg CPM by two 0.25 ml intraperitoneal injections scheduled on day 0 (150 mg/kg) and day 3 (100 mg/kg). 15 On day 3, some mice were subcutaneously treated with 0.2 ml of T140 (5 mg/kg), G-CSF (5 mg/mouse) or their combination once a day for five constitutive days (day 3-7). BM and blood samples were taken on days 0, 3, 4, 5, 6 and 7 ( Figure 2a).…”

Section: Mobilization With T140 and Cyclophosphamide Treatmentmentioning

confidence: 99%

The CXCR4 antagonist 4F-benzoyl-TN14003 stimulates the recovery of the bone marrow after transplantation

Abraham¹,

Beider²,

Wald³

et al. 2009

Leukemia

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Cytopenia represents a significant complication after chemotherapy, irradiation before bone marrow (BM) transplantation or as a therapy for cancer. The mechanisms that determine the pace of BM recovery are not fully understood. During the recovery phase after chemotherapy or irradiation, the signals for retention of white blood cells within the BM increase significantly. This leads to a delay in the release of WBC, which can be overcome by targeting the CXCR4 axis with the antagonist 4F-benzoyl-TN14003 (T140). The delay in the release of WBC is also accompanied by suppression in the production of progenitor cells and mature cells by the BM stroma. Administration of T140 to mice transplanted with BM cells stimulates the production of all types of progenitors and mature cells, and increases the exit of mature cells to the periphery. Moreover, addition of T140, but not AMD3100, to BM stromal cultures stimulates the production of mature cells and progenitors from all lineages. The unique ability of the CXCR4 antagonist, T140 to stimulate the production and exit of WBC cells may be used as a novel therapeutic approach to overcome cytopenia associated with treatments for cancer and BM transplantation.

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Neutropenia induced in outbred mice by a simplified low-dose cyclophosphamide regimen: characterization and applicability to diverse experimental models of infectious diseases

Cited by 165 publications

References 24 publications

Neutrophil Depletion Causes a Fatal Defect in Murine Pulmonary Staphylococcus aureus clearance

Neutrophil Depletion Causes a Fatal Defect in Murine Pulmonary Staphylococcus aureus clearance

Absolute counting of neutrophils in whole blood using flow cytometry

The CXCR4 antagonist 4F-benzoyl-TN14003 stimulates the recovery of the bone marrow after transplantation

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