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• W2550271193 abstract "1142 Objectives Determine LFA-1 receptor-binding characteristics of radiolabeled DANBIRT using radioisotopic dilution methods in isolated leukocytes from control and ozone-exposed rats using 68Ga-DANBIRT and 111In-DANBIRT. Methods All experiments were done using Sprague Dawley male rats. For the first experiment, rats were exposed to filtered air (n=4) and ozone [1 ppm (n=4)] for 4 hours. Whole blood was collected 24 hours post-exposure and incubated for 10 minutes with 68Ga-DANBIRT. For the second experiment, radioisotopic dilution methods were performed with 1:1, 1:10, 1:100 and 1:1000 log-fold serial dilution concentrations of DANBIRT. Whole blood was collected and incubated for 1 hour with 111In-DANBIRT. Leukocyte isolation was performed for neutrophil and peripheral blood mononuclear cell (PBMC) separations post-incubation with radiolabeled DANBIRT (Standard Specific Activity of 625mCi/pM). Cell morphology and sample purity were assessed by light microscopy. Sample smears and cytospin slides were stained using Wright-Giemsa. Results Leukocyte isolation methods successfully showed the purity of samples, with predominance of immature neutrophils (65%) and PBMCs (58%) in ozone-exposed rats with no morphologic changes. In vivo data showed an increased uptake of 68Ga-DANBIRT in neutrophils with a decreased uptake in PBMCs post-ozone exposure. Radioisotopic dilution methods evidence a saturation of binding sites in our isolated neutrophil samples starting at a 1:10 log-fold serial dilution from our initial concentration of 111In-DANBIRT (Figure 1). Conclusions Neutrophil and PBMC isolation methods helped demonstrate competitive receptor binding in leukocytes; showing an increased percentage of immature neutrophils, which illustrates an acute immune response in the ozone exposed group. Radiosotopic dilution methods evidence competitive binding in leukocytes. Competitive binding is increased when concentration increases, due to a mass effect of radiolabeled DANBIRT at equal or higher concentrations of 6.36e5pM. Previous studies using 111In-DANBIRT have demonstrated its potential as a SPECT/CT molecular imaging probe capable of visualizing inflammation found in models of cardiovascular injury. These correlative results demonstrate competitive and specific LFA-1 receptor binding especially in neutrophils, which will guide future studies using competitive binding concentrations in other vascular injury models.RESEARCH SUPPORT: Data was generated in the Keck-UNM Small-Animal Imaging Resource Shared Resource Center supported by the University of New Mexico Health Sciences Center, the UNM Cancer Center, and UNM College of Pharmacy. Grant numbers: NIH P30CA118100-06 & ES0014639." @default.
• W2550271193 created "2016-11-30" @default.
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• W2550271193 date "2016-05-01" @default.
• W2550271193 modified "2023-09-29" @default.
• W2550271193 title "Competitive receptor-binding assays of 111In-DANBIRT targeting of Leukocyte-function associated antigen-1 in a systemic inflammation rat model to inhaled ozone exposure." @default.
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