Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of human polymorphonuclear leukocytes (PMNs) from healthy individuals vs patients with X-linked chronic granulomatous disease


ABSTRACT: Polymorphonuclear leukocytes (PMNs) were obtained from healthy individuals (Healthy1, Healthy2, Healthy3, and Healthy4) or patients with X-linked chronic granulomatous disease (XCGD1, XCGD2, XCGD3, XCGD4, XCGD5, and XCGD6). The studies were performed in accordance with a protocol approved by the Institutional Review Board for Human Subjects, National Institute of Allergy and Infectious Diseases, and the Institutional Review Board for Human Subjects at the University of Iowa. All studies were conducted according to Declaration of Helsinki principles. PMNs (107) were combined with or without IgG and C3bi-coated latex beads (8 x 107) in wells of a 12-well tissue culture plate (pre-coated with 20% normal human serum) and centrifuged at 350 x g for 8 min at 4°C to synchronize phagocytosis. For the purpose of these studies, activated or Stimulated PMNs are defined as those that have been stimulated by phagocytosis of IgG- and C3bi-coated latex beads. Control PMNs are defined as resting cells or those left unstimulated throughout the time-course. Following centrifugation, plates were incubated at 37 °C in a CO2 for the indicated times. At the indicated times, tissue culture medium was aspirated from the plate and PMNs were lysed directly with RLT buffer (Qiagen, Valencia, CA). Purification of PMN RNA and subsequent preparation of labeled cRNA target (12 g) was performed as described in Methods. Labeling of samples, hybridization of cRNA with Hu95Av2 oligonucleotide arrays (Affymetrix, Santa Clara, CA), and scanning were performed according to standard Affymetrix protocols. Gene expression in healthy control and XCGD individuals was always compared at the same time points after phagocytosis

ORGANISM(S): Homo sapiens

DISEASE(S): XCGD

SUBMITTER: Frank DeLeo 

PROVIDER: E-GEOD-935 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Gene expression profiling provides insight into the pathophysiology of chronic granulomatous disease.

Kobayashi Scott D SD   Voyich Jovanka M JM   Braughton Kevin R KR   Whitney Adeline R AR   Nauseef William M WM   Malech Harry L HL   DeLeo Frank R FR  

Journal of immunology (Baltimore, Md. : 1950) 20040101 1


Human polymorphonuclear leukocytes (PMNs or neutrophils) kill invading microorganisms with reactive oxygen species (ROS) and cytotoxic granule components. PMNs from individuals with X-linked chronic granulomatous disease (XCGD) do not produce ROS, thereby rendering these individuals more susceptible to infection. In addition, XCGD patients develop tissue granulomas that obstruct vital organs, the mechanism(s) for which are unknown. To gain insight into the molecular processes that contribute to  ...[more]

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