Immunophenotyping of Human B Lymphocytes in Blood and in Adipose Tissue

  • Alain Diaz
  • Maria Romero
  • Daniela Frasca
  • Bonnie B. BlombergEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2032)


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.

Key words

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) Red blood cells (RBC) Reactive oxygen species (ROS) Room temperature (RT) Subcutaneous adipose tissue (SAT) Stromal vascular fraction (SVF) Type-2 diabetes (T2D) Toll-like receptor (TLR) 



This study was supported by NIH R56 AG32576 (DF/BBB), R21 AG042826 (DF/BBB), R21 AI096446 (BBB/DF), R56 AG059719 [4].


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Alain Diaz
    • 1
  • Maria Romero
    • 1
  • Daniela Frasca
    • 1
  • Bonnie B. Blomberg
    • 1
    • 2
    Email author
  1. 1.Department of Microbiology and ImmunologyUniversity of Miami Miller School of MedicineMiamiUSA
  2. 2.Sylvester Comprehensive Cancer CenterUniversity of Miami Miller School of MedicineMiamiUSA

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