Effects of anticoagulants and Ficoll on human serum antibody reactivities and functions against Mycobacterium tuberculosis

Tuberculosis (Edinb). 2020 Jan:120:101901. doi: 10.1016/j.tube.2020.101901. Epub 2020 Jan 10.

Abstract

The ability to utilize leftover samples containing anticoagulants or Ficoll would provide substantial opportunities for future antibody and biomarker studies. Some anticoagulants might influence antibody reactivity against pathogens, but comprehensive studies investigating effects in the context of TB are lacking. We enrolled 24 individuals with and without history of M. tuberculosis and/or HIV-infection and investigated TB antibody reactivities, function, and other host protein biomarkers in simultaneously obtained serum and plasma from serum separation, EDTA, heparin, acid citrate dextrose (ACD), or mononuclear cell preparation (CPT™) tubes which contain heparin and Ficoll. Antibody isotype reactivities to two mycobacterial antigens, as well as phagocytosis of M. tuberculosis, correlated strongly and significantly between serum and plasma, irrespective of type of anticoagulant or Ficoll present (r ≥ 0.85, p < 0.0001). However, the presence of ACD resulted in slightly lower values than those obtained with serum in both indirect (antibody reactivities to mycobacterial antigens) and Sandwich ELISAs (soluble CD14 measurements). Our data demonstrate that leftover plasma, regardless of containing anticoagulants or Ficoll, can be used in TB antibody or other host protein biomarker studies but suggest the value of a correction factor when using ACD plasma interchangeably with serum in antibody binding studies.

Keywords: ACD; CPT; EDTA; Heparin; Immunoglobulins; Proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Antibodies, Bacterial / blood
  • Antibodies, Bacterial / drug effects*
  • Antibody Specificity / drug effects
  • Anticoagulants / pharmacology*
  • Antigens, Bacterial / immunology
  • Binding Sites, Antibody
  • Blood Specimen Collection
  • Citric Acid / pharmacology
  • Coinfection
  • Edetic Acid / pharmacology
  • Female
  • Ficoll / pharmacology*
  • Glucose / analogs & derivatives
  • Glucose / pharmacology
  • HIV Infections / blood
  • HIV Infections / immunology
  • HIV Infections / virology
  • Heparin / pharmacology
  • Humans
  • Macrophages / drug effects
  • Macrophages / immunology
  • Macrophages / microbiology
  • Male
  • Middle Aged
  • Mycobacterium tuberculosis / immunology*
  • Phagocytosis / drug effects
  • THP-1 Cells
  • Tuberculosis / blood
  • Tuberculosis / immunology*
  • Tuberculosis / microbiology

Substances

  • Antibodies, Bacterial
  • Anticoagulants
  • Antigens, Bacterial
  • acid citrate dextrose
  • Ficoll
  • Citric Acid
  • Heparin
  • Edetic Acid
  • Glucose