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Isolation of Leukocytes from Human Breast Milk for Use in an Antibody-dependent Cellular Phagocytosis Assay of HIV Targets
Published on: September 6, 2019
Cytokine production by human milk cells and peripheral blood mononuclear cells from the same mothers
Joanna S Hawkes1, Dani-Louise Bryan, Robert A Gibson
1Child Nutrition Research Centre, Flinders Medical Centre, Bedford Park, South Australia, Australia.
Samples of milk (n = 80) and venous blood were collected at 5 weeks postpartum from 82 lactating mothers. Human milk cells and peripheral blood mononuclear cells were isolated and the production of interleukin-1beta, interleukin-6, and tumor necrosis factor-a in the absence and presence of lipopolysaccharide was determined by enzyme-linked immunosorbent assay. Human milk cells spontaneously produced significantly less interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha than peripheral blood mononuclear cells in the absence of stimulation. In vitro stimulation of human milk cells with lipopolysaccharide (500 ng/ml) for 24 hr increased cytokine production by approximately 40-50%, whereas peripheral blood mononuclear cells responded to lipopolysaccharide (200 ng/ml) with increased cytokine production of up to 350%. These observations suggest that cells in milk are capable of active involvement in the production of the interleukin-1beta, interleukin-6, and tumor necrosis factor-a in the mammary gland and have the capacity to respond to further stimulation after leaving the breast.
Samples of milk (n = 80) and venous blood were collected at 5 weeks postpartum from 82 lactating mothers. Human milk cells and peripheral blood mononuclear cells were isolated and the production of interleukin-1beta, interleukin-6, and tumor necrosis factor-a in the absence and presence of lipopolysaccharide was determined by enzyme-linked immunosorbent assay. Human milk cells spontaneously produced significantly less interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha than peripheral blood mononuclear cells in the absence of stimulation. In vitro stimulation of human milk cells with lipopolysaccharide (500 ng/ml) for 24 hr increased cytokine production by approximately 40-50%, whereas peripheral blood mononuclear cells responded to lipopolysaccharide (200 ng/ml) with increased cytokine production of up to 350%. These observations suggest that cells in milk are capable of active involvement in the production of the interleukin-1beta, interleukin-6, and tumor necrosis factor-a in the mammary gland and have the capacity to respond to further stimulation after leaving the breast.
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