Evaluation of immunomodulatory effect of recombinant human granulocyte‐macrophage colony‐stimulating factor on polymorphonuclear cell from dogs with cancer in vitro

Abstract

The objective of this in vitro study was to evaluate the immunomodulatory effects of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) on polymorphonuclear cell (PMN) function in dogs with cancer. PMNs were harvested from dogs with naturally developing cancer as a pre-clinical model to evaluate the immunomodulatory effects of rhGM-CSF on PMN phagocytic and cytotoxic functions, cytokine production and receptor expression. Some aspects of cancer-related PMN dysfunction in dogs with cancer were restored following incubation with rhGM-CSF including PMN phagocytosis, respiratory burst and LPS-induced TNF-α production. In addition, rhGM-CSF increased surface HLA-DR expression on the PMNs of dogs with cancer. These data suggests that dysfunction of innate immune response in dogs with cancer may be improved by rhGM-CSF. The results of this study provided a pathophysiologic rationale for the initiation of clinical trials to continue evaluating rhGM-CSF as an immunomodulatory therapy in dogs with cancer.

Immune Function in Critically Ill Dogs

“…A CyAn ADP flow cytometry instrument (Beckman Coulter, Inc.; Brea, CA, USA) and associated data analysis software (Summit V5.2.0.7477, Dako Colorado, Inc.; Ft. Collins, CO, USA) were used for analysis of phagocytosis and respiratory burst capacity in phagocytic cells, including neutrophils and monocytes, quantification of CD3 + /CD4 + , CD3 + /CD8 + , and CD21 + lymphoid phenotypes, and quantification of monocytic cells expressing HLA‐DR and TLR‐4 for all the dogs enrolled in this study using previously described methods …”

“…A CyAn ADP flow cytometry instrument (Beckman Coulter, Inc.; Brea, CA, USA) and associated data analysis software (Summit V5.2.0.7477, Dako Colorado, Inc.; Ft. Collins, CO, USA) were used for analysis of phagocytosis and respiratory burst capacity in phagocytic cells, including neutrophils and monocytes, quantification of CD3 + /CD4 + , CD3 + /CD8 + , and CD21 + lymphoid phenotypes, and quantification of monocytic cells expressing HLA-DR and TLR-4 for all the dogs enrolled in this study using previously described methods. 14,[17][18][19] For phagocytosis and respiratory burst function, aggregation artifacts caused by bacteria or cells were excluded by gating around live cells based on DNA stain uptake (Fig 1). Gating of phagocytic cells, including both neutrophils and monocytes, was performed using forward versus side scatter.…”

Dogs with osteosarcoma have altered pro‐ and anti‐inflammatory cytokine profiles

“…While inflammation and cytokine production in dogs with cancer is not well studied, particularly in dogs with OSA, one study found that dogs with OSA had increased plasma TNF concentrations compared to healthy dogs (Tuohy, Lascelles, Griffith, & Fogle, ). Other studies investigated leukocyte cytokine secretion in dogs with lymphoma and soft tissue sarcoma (Axiak‐Bechtel et al., ; Fowler, Axiak, & DeClue, ; Zhang et al., ), but these immunologic parameters have not been investigated in dogs with OSA. The objective of this study was to determine the leukocyte production of TNF, IL‐6 and IL‐10 in addition to the TNF and IL‐6 to IL‐10 ratios at the time of OSA diagnosis following stimulation with pathogen‐associated molecular pathogens.…”

“…Other studies investigated leukocyte cytokine secretion in dogs with lymphoma and soft tissue sarcoma (Axiak-Bechtel et al, 2014;Fowler, Axiak, & DeClue, 2011;Zhang et al, 2017), but these immunologic parameters have not been investigated in dogs with OSA.…”

Identification of immunologic and clinical characteristics that predict inflammatory response to C. Novyi-NT bacteriolytic immunotherapy

“…Proposed explanations for the limited response rate include a lack of appropriate immune response to infection or variability in the degree of hypoxic tissue in the tumor leading to reduced CNV-NT germination. Dogs with various forms of neoplasia have altered immune function which could impact the efficacy of immunotherapies like induced infection and little is known about the regional blood flow distribution in solid tumors in this species [ 5 – 8 ].…”