Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue

Alain Diaz; Maria Romero; Daniela Frasca; Bonnie B. Blomberg

doi:10.1007/978-1-4939-9650-6_7

Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue

Alain Diaz, Maria Romero, Daniela Frasca, Bonnie B. Blomberg

Microbiology & Immunology

Research output: Contribution to journal › Article

1 Scopus citations

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	https://doi.org/10.1007/978-1-4939-9650-6_7
State	Published - Jan 1 2019

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

Access to Document

10.1007/978-1-4939-9650-6_7

Fingerprint Dive into the research topics of 'Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue'. Together they form a unique fingerprint.

Cite this

Diaz, A., Romero, M., Frasca, D., & Blomberg, B. B. (2019). Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue. Methods in molecular biology (Clifton, N.J.), 2032, 115-127. https://doi.org/10.1007/978-1-4939-9650-6_7

@article{5df7e90c87c74f5e88d55fb1dc07c3e8,

title = "Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue",

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.",

keywords = "Adipose tissue (AT), Body mass index (BMI), Cardiovascular (CV), Dulbecco{\textquoteright}s modified Eagle{\textquoteright}s medium (DMEM), Fetal calf serum (FCS), Free fatty acids (FFAs), Germinal center (GC), Hanks{\textquoteright} 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)",

author = "Alain Diaz and Maria Romero and Daniela Frasca and Blomberg, {Bonnie B.}",

year = "2019",

month = jan,

day = "1",

doi = "10.1007/978-1-4939-9650-6_7",

language = "English (US)",

volume = "2032",

pages = "115--127",

journal = "Methods in molecular biology (Clifton, N.J.)",

issn = "1064-3745",

publisher = "Humana Press",

}

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 -