+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Flow cytometry complement-mediated cytotoxicity assay detects baboon xenoantibodies directed to porcine epitopes undetected by hemolytic assay

Flow cytometry complement-mediated cytotoxicity assay detects baboon xenoantibodies directed to porcine epitopes undetected by hemolytic assay

Transplant Immunology 13(4): 313-317

The pig-to-primate model is increasingly being utilized as the final preclinical means of assessing therapeutic strategies aimed at allowing discordant xenotransplantation. To obtain information about the nature of cytotoxic response in pig-to-baboon xenotransplants, we sought to determine if serum cytotoxicity in this model was assay dependent. Sera from nine kidney or heart xenotransplanted baboons were obtained before transplantation and at the time of acute humoral xenograft rejection (AHXR). Cytotoxicity was measured by an anti-pig haemolytic assay (APHA) and by a flow cytometry complement-dependent assay (FCCA), using pig blood lymphocytes (PBLs). Serum samples showing inter-assay differences were absorbed with pig erythrocytes and assayed by APHA and FCCA, as well as by measuring anti-alphaGal and total anti-pig xenoantibodies. The results showed that in four AHXR samples, FCCA cytotoxicity was higher than APHA cytotoxicity. Absorption with pig erythrocytes diminished FCCA and removed APHA cytotoxicity. Residual FCCA activity was due to total anti-pig and IgM anti-alphaGal and non-Gal antibodies. Our results indicate that some cytotoxic antibodies present in the sera of xenotransplanted baboons at time of AHXR are IgM antibodies directed against pig PBL antigens not detected by APHA.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 049092120

Download citation: RISBibTeXText

PMID: 15589745

DOI: 10.1016/j.trim.2004.09.001

Related references

FDA/PI flow cytometry assay of complement-mediated cytotoxicity of antibodies generated during xenotransplantation. Cytometry. Part A 62(1): 54-60, 2004

An improved fluorescence assay for the determination of lymphocyte-mediated cytotoxicity using flow cytometry. Journal Of Immunological Methods. 177(1-2): 101-111, 1994

Rapid flow cytometry-based assay for the evaluation of γδ T cell-mediated cytotoxicity. Molecular Medicine Reports 17(3): 3555-3562, 2018

Antibody-dependent cell-mediated cytotoxicity: A flow cytometry-based assay using fluorophores. Journal of Immunological Methods 258(1-2): 183-191, 1 December, 2001

A novel multiparametric flow cytometry-based cytotoxicity assay simultaneously immunophenotypes effector cells: comparisons to a 4 h 51Cr-release assay. Journal of Immunological Methods 325(1-2): 51-66, 2007

Two modifications of the clonogenic assay: One which detects cell mediated cytotoxicity and a second for factor mediated cytotoxicity. Methods in Cell Science 13(2): 133-136, 1991

A flow cytometry based assay that simultaneously measures cytotoxicity and monocyte mediated antibody dependent effector activity. Journal of Immunological Methods 462: 74-82, 2018

A flow cytometry-based assay for the evaluation of antibody-dependent cell-mediated cytotoxicity (ADCC) in cancer cells. Methods in Molecular Biology 1165: 241-252, 2014

A differential assay of NK-cell-mediated cytotoxicity in K562 cells revealing three sequential membrane impairment steps using three-color flow-cytometry. Journal of Immunological Methods 307(1-2): 41-53, 2005

A flow cytometry assay simultaneously detects independent apoptotic parameters. Cytometry 45(2): 151-157, 2001

Hemolytic assay for goat (caprine) and swine (porcine) complement. Veterinary Immunology and Immunopathology 1(4): 317-328, 1980

Detection of antibody-dependent cell-mediated cytotoxicity by automated flow cytometry. Comparison to a chromium release assay and characterization of the effector cell subpopulation. Journal of Immunological Methods 36(3-4): 325-333, 1980

Fluorescent target array T helper assay: a multiplex flow cytometry assay to measure antigen-specific CD4+ T cell-mediated B cell help in vivo. Journal of Immunological Methods 387(1-2): 181-190, 2013

Flow cytometry-based cytotoxicity and antibody binding assay. Current Protocols in Cytometry 66: 6.34.1-6.34.11, 2013

A novel assay for assessment of HIV-specific cytotoxicity by multiparameter flow cytometry. Cytometry. Part A 68(2): 71-80, 2005