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32. Donor antibodies to HNA-3a implicated in TRALI reactions prime neutrophils and cause PMN-mediated damage to human pulmonary microvascular endothelial cells in a two-event in vitro model

Author

Silliman, Christopher C., Curtis, Brian R., Kopko, Patricia M., Khan, Samina Y., Kelher, Marguerite R., Schuller, Randy M., Sannoh, Baindu, and Ambruso, Daniel R.

Abstract

Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-related mortality. Antibodies to HNA-3a are commonly implicated in TRALI. We hypothesized that HNA-3a antibodies prime neutrophils (PMNs) and cause PMN-mediated cytotoxicity through a two-event pathogenesis. Isolated HNA-3a+or HNA-3a−PMNs were incubated with plasma containing HNA-3a antibodies implicated in TRALI, and their ability to prime the oxidase was measured. Human pulmonary microvascular endothelial cells (HMVECs) were activated with endotoxin or buffer, HNA-3a+or HNA-3a−PMNs were added, and the coculture was incubated with plasma ± antibodies to HNA-3a. PMN-mediated damage was measured by counting viable HMVECs/mm2. Plasma containing HNA-3a antibodies primed the fMLP-activated respiratory burst of HNA-3a+, but not HNA-3a−, PMNs and elicited PMN-mediated damage of LPS-activated HMVECs when HNA-3a+, but not HNA-3a−, PMNs were used. Thus, antibodies to HNA-3a primed PMNs and caused PMN-mediated HMVEC cytotoxicity in a two-event model identical to biologic response modifiers implicated in TRALI.

Published
2007
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33. Antithrombin III Deficiency: Decreased Synthesis of a Biochemically Normal Molecule

Author

Ambruso, Daniel R., Leonard, Bruce D., Bies, Roger D., Jacobson, Linda, Hathaway, Wm.E., and Reeve, E.Basil

Abstract

A 29-yr-old white female has suffered from recurrent venous thromboses over the last 12 yr. Plasma antithrombin III (AT-III) levels were 48% of normal by Immunoelectrophoresis and 56% by chromogenic assay. Three of four siblings and the father had similar AT-III levels without associated venous thromboses. Heparin-Sepharose chromatography demonstrated normal behavior of the patient's AT-III. Her purified AT-III could not be distinguished from AT-III purified from a normal control either by SDS Polyacrylamide gel electrophoresis or by crossed Immunoelectrophoresis, and the heparin cofactor activity and the progressive antithrombin activity of both AT-III samples were identical. Turnover studies were made in the patient using her own purified AT-III labeled with 131I, (*l). The results did not differ significantly from studies made with autologous *I-AT-III in two normal control women. Her fractional breakdown rate of 0.54 total plasma AT-III per day compared with 0.45 and 0.52 in the controls. These studies indicate that the patient synthesizes a normal AT-III molecule at half normal rates.

Published
1982
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34. In VivoEffects of Interferon-Gamma 1-b (IFN-γ) on Neutrophils

Author

Ambruso, Daniel R., Ellison, Michael A, Briones, Natalie, and Baroffio, Angelina

Abstract

Introduction: Most information about the immunoregulatory functions of INF-γ has focused on the interaction with the adaptive immune system and less is reported about neutrophil function. Furthermore, the majority of studies with neutrophils evaluate the effects of INF-γ on differentiated cells. The clinical use of IFN-γ has been driven by these data and the clinical findings that administration of this cytokine to patients with Chronic Granulomatous Disease results in decreased incidence of severe infections although the defect in Nox2 activity is not altered. To determine the in vivoeffects on myeloid cells developed under the influence of this cytokine, we evaluated neutrophils from healthy human subjects receiving IFN-γ.

Published
2016
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