Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue

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

Research output: Chapter in Book/Report/Conference proceedingChapter

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)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages115-127
Number of pages13
DOIs
StatePublished - 2019
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume2032
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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

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