TY - CHAP
T1 - Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue
AU - Diaz, Alain
AU - Romero, Maria
AU - Frasca, Daniela
AU - Blomberg, Bonnie B.
N1 - Funding Information:
This study was supported by NIHR56 AG32576 (DF/BBB), R21 AG042826 (DF/BBB), R21 AI096446 (BBB/DF), R56 AG059719 [4].
Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2019.
PY - 2019
Y1 - 2019
N2 - The human obese subcutaneous adipose tissue (SAT) contributes to systemic and B cell intrinsic inflammation, reduced B cell responses, and increased secretion of autoimmune antibodies. Immune cells are recruited to the SAT by chemokines released by both adipocytes and infiltrating immune cells. We describe here the characterization of B lymphocytes from the SAT and blood (control) of obese females undergoing weight reduction surgeries (breast reduction or panniculectomy). We show how to isolate the immune cells from the blood and SAT, how to characterize B cells and their subsets, and how to measure markers of activation and/or transcription factors in SAT-derived B cells and B cell subsets. We also show how to evaluate other immune cell types infiltrating the SAT, including T cells, NK cells, monocyte/macrophages, in order to measure relative proportions of these cell types as compared to the blood.
AB - The human obese subcutaneous adipose tissue (SAT) contributes to systemic and B cell intrinsic inflammation, reduced B cell responses, and increased secretion of autoimmune antibodies. Immune cells are recruited to the SAT by chemokines released by both adipocytes and infiltrating immune cells. We describe here the characterization of B lymphocytes from the SAT and blood (control) of obese females undergoing weight reduction surgeries (breast reduction or panniculectomy). We show how to isolate the immune cells from the blood and SAT, how to characterize B cells and their subsets, and how to measure markers of activation and/or transcription factors in SAT-derived B cells and B cell subsets. We also show how to evaluate other immune cell types infiltrating the SAT, including T cells, NK cells, monocyte/macrophages, in order to measure relative proportions of these cell types as compared to the blood.
KW - Adipose tissue (AT)
KW - Body mass index (BMI)
KW - Cardiovascular (CV)
KW - Dulbecco’s modified Eagle’s medium (DMEM)
KW - Fetal calf serum (FCS)
KW - Free fatty acids (FFAs)
KW - Germinal center (GC)
KW - Hanks’ balanced salt solution (HBSS)
KW - Insulin resistance (IR)
KW - Insulin sensitivity (IS)
KW - Peripheral blood mononuclear cells (PBMC)
KW - Reactive oxygen species (ROS)
KW - Red blood cells (RBC)
KW - Room temperature (RT)
KW - Stromal vascular fraction (SVF)
KW - Subcutaneous adipose tissue (SAT)
KW - Toll-like receptor (TLR)
KW - Type-2 diabetes (T2D)
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U2 - 10.1007/978-1-4939-9650-6_7
DO - 10.1007/978-1-4939-9650-6_7
M3 - Chapter
C2 - 31522416
AN - SCOPUS:85072168435
T3 - Methods in Molecular Biology
SP - 115
EP - 127
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -