Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue

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

Research output: Contribution to journalArticle

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 languageEnglish (US)
Pages (from-to)115-127
Number of pages13
JournalMethods in molecular biology (Clifton, N.J.)
Volume2032
DOIs
StatePublished - Jan 1 2019

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Immunophenotyping
Subcutaneous Fat
Adipose Tissue
B-Lymphocytes
B-Lymphocyte Subsets
Abdominoplasty
Chemokines
Adipocytes
Natural Killer Cells
Weight Loss
Monocytes
Blood Cells
Breast
Transcription Factors
Macrophages
Inflammation
T-Lymphocytes
Antibodies

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 journalArticle

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