American Society of Hematology

Figure 2.

$Figure 2. Anti-HNA-2a leads to neutrophil activation and clustering of leukocytes in the capillaries of the lung. (A) Production of ROS as determined by flow cytometry. Left panel shows response to mAb 7D8; right panel shows response to IgG fractions obtained from 2 of 3 analyzed human sera containing anti-HNA-2a. Appropriate controls (murine and human IgG, respectively) are given as histograms with dotted lines. All histograms are representative of 3 independent experiments. (B,C) Clustering of leukocytes in pulmonary capillaries following the administration of anti-HNA-2a (B), but not of control antibody (C). After the final venous pressure elevation, rat lungs were instilled with formaldehyde solution (4.5%, pH 7.2) through the trachea. Fixation was allowed to proceed overnight at room temperature. Subsequently, tissue samples were embedded in paraffin and sections of 5 μm from all lungs were stained with hematoxylin-eosin and antimyeloperoxidase. Pictures are representative for experiments and controls (antimyeloperoxidase, APAAP). A Zeiss Axioskop 40 (Carl Zeiss, Jena, Germany) with Plan-NEO Fluar 20×/0.5 NA objective lenses, in connection with a JVC KY-S75U digital camera (JVC, Friedberg, Germany) was used to acquire the micrographs. Images were processed with Diskus acquisition software, version 4.50 (Hilgers, Königswinter, Germany).$

Anti-HNA-2a leads to neutrophil activation and clustering of leukocytes in the capillaries of the lung. (A) Production of ROS as determined by flow cytometry. Left panel shows response to mAb 7D8; right panel shows response to IgG fractions obtained from 2 of 3 analyzed human sera containing anti-HNA-2a. Appropriate controls (murine and human IgG, respectively) are given as histograms with dotted lines. All histograms are representative of 3 independent experiments. (B,C) Clustering of leukocytes in pulmonary capillaries following the administration of anti-HNA-2a (B), but not of control antibody (C). After the final venous pressure elevation, rat lungs were instilled with formaldehyde solution (4.5%, pH 7.2) through the trachea. Fixation was allowed to proceed overnight at room temperature. Subsequently, tissue samples were embedded in paraffin and sections of 5 μm from all lungs were stained with hematoxylin-eosin and antimyeloperoxidase. Pictures are representative for experiments and controls (antimyeloperoxidase, APAAP). A Zeiss Axioskop 40 (Carl Zeiss, Jena, Germany) with Plan-NEO Fluar 20×/0.5 NA objective lenses, in connection with a JVC KY-S75U digital camera (JVC, Friedberg, Germany) was used to acquire the micrographs. Images were processed with Diskus acquisition software, version 4.50 (Hilgers, Königswinter, Germany).

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