TY - JOUR
T1 - Effects of anticoagulants and Ficoll on human serum antibody reactivities and functions against Mycobacterium tuberculosis
AU - Gadsden, Nicholas J.
AU - Liu, Yanyan
AU - Gati, Stephanie
AU - Chen, Tingting
AU - Jenny-Avital, Elizabeth R.
AU - Achkar, Jacqueline M.
N1 - Funding Information:
This work was supported in part by grants from the National Institutes of Health (NIH/NIAID) AI117927 , AI127173 , and AI146329 to J.M.A. All authors declared no conflict of interest. The following antigens and murine mAbs were obtained through BEI Resources, NIAID, NIH: native PstS1 (Rv0934), purified from M. tuberculosis , Strain H37Rv, NR-14859; mAb to PhoS 1/PstS1 (IT-15, NR-13605); and mAb to LAM (CS-35, NR-13811). Appendix A
Funding Information:
This work was supported in part by grants from the National Institutes of Health (NIH/NIAID) AI117927, AI127173, and AI146329 to J.M.A. All authors declared no conflict of interest. The following antigens and murine mAbs were obtained through BEI Resources, NIAID, NIH: native PstS1 (Rv0934), purified from M. tuberculosis, Strain H37Rv, NR-14859; mAb to PhoS 1/PstS1 (IT-15, NR-13605); and mAb to LAM (CS-35, NR-13811).
Publisher Copyright:
© 2020
PY - 2020/1
Y1 - 2020/1
N2 - 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.
AB - 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.
KW - ACD
KW - CPT
KW - EDTA
KW - Heparin
KW - Immunoglobulins
KW - Proteins
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U2 - 10.1016/j.tube.2020.101901
DO - 10.1016/j.tube.2020.101901
M3 - Article
C2 - 32090862
AN - SCOPUS:85077736809
SN - 1472-9792
VL - 120
JO - Tuberculosis
JF - Tuberculosis
M1 - 101901
ER -
