Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 115-127 |
| Number of pages | 13 |
| Journal | Methods in molecular biology (Clifton, N.J.) |
| Volume | 2032 |
| DOIs | |
| State | Published - Jan 1 2019 |
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Keywords
- Adipose tissue (AT)
- Body mass index (BMI)
- Cardiovascular (CV)
- Dulbecco’s modified Eagle’s medium (DMEM)
- Fetal calf serum (FCS)
- Free fatty acids (FFAs)
- Germinal center (GC)
- Hanks’ balanced salt solution (HBSS)
- Insulin resistance (IR)
- Insulin sensitivity (IS)
- Peripheral blood mononuclear cells (PBMC)
- Reactive oxygen species (ROS)
- Red blood cells (RBC)
- Room temperature (RT)
- Stromal vascular fraction (SVF)
- Subcutaneous adipose tissue (SAT)
- Toll-like receptor (TLR)
- Type-2 diabetes (T2D)
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue. / Diaz, Alain; Romero, Maria; Frasca, Daniela; Blomberg, Bonnie B.
In: Methods in molecular biology (Clifton, N.J.), Vol. 2032, 01.01.2019, p. 115-127.Research output: Contribution to journal › Article
}
TY - JOUR
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.
PY - 2019/1/1
Y1 - 2019/1/1
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)
UR - http://www.scopus.com/inward/record.url?scp=85072168435&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85072168435&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-9650-6_7
DO - 10.1007/978-1-4939-9650-6_7
M3 - Article
C2 - 31522416
AN - SCOPUS:85072168435
VL - 2032
SP - 115
EP - 127
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
SN - 1064-3745
ER -