title: Comparison of several functional methods to evaluate the immune response on stable kidney transplant patients.
abstract: The introduction of new immunosuppressive drugs in the last two decades has been associated with a significant decline in the prevalence of acute rejection and a huge improvement of graft survival. Monitoring blood levels of immunosuppressive drugs is the most common way to control drug doses in renal transplant patients. This approach is useful and widely used but doesn't give accurate information about the immune status of the patient. For this goal, there are many "in house" protocols which give more information, but cannot be standardized, limiting their applicability to compare results between different laboratories. In this study we compare three classical functional methods to evaluate the immune response: Mixed lymphocyte reaction (MLR), phytohemagglutinin stimulated peripheral blood lymphocytes (PBLs), and anti-CD3 monoclonal antibodies (mAbs) against PBL with the only FDA-labeled assay to measure the patient immune status: Cylex ImmuKnow® that measures the intracelullar ATP in CD4+ lymphocytes. We used n=111 stable renal transplant patients, all the patients with more than one year functioning grafts. We referred the results to a control population of healthy blood donors (n=125).
  
  
  RESULTS
  Measurement of intracellular ATP in CD4+ lymphocytes is able to differentiate immunosuppressed populations in renal transplant patients from health controls (242.30±21.62 vs. 386.43±25.12, p 0.0001). By contrary, there were no differences between controls and renal recipients when functional response was measured by MLR, PHA and anti-CD3 mAbs (2.48±0.45 vs. 2.37±0.41; 2.84±0.76 vs. 2.37±0.32; 2.32±0.34 vs. 1.89±0.38 respectively). In summary, our results show that the measurement of ATP in CD4+ lymphocytes gives more accurate information in comparison to the classical methods.
authors.affiliation: Department of Immunology, Instituto de Investigación Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: José A
authors.initials: JA
authors.lastname: Martínez-Flores
authors.affiliation: Department of Nephrology, Instituto de Investigación Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Manuel
authors.initials: M
authors.lastname: Serrano
authors.affiliation: Department of Immunology, Instituto de Investigación Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: Pablo
authors.initials: P
authors.lastname: Morales
authors.affiliation: Department of Immunology, Instituto de Investigación Hospital Universitario 12 de Octubre, Madrid, Spain; Section of Immunology, Universidad San Pablo-CEU, Madrid, Spain.
authors.forename: Estela
authors.initials: E
authors.lastname: Paz-Artal
authors.affiliation: Department of Nephrology, Instituto de Investigación Hospital Universitario 12 de Octubre, Madrid, Spain.
authors.forename: José M
authors.initials: JM
authors.lastname: Morales
authors.affiliation: Department of Immunology, Instituto de Investigación Hospital Universitario 12 de Octubre, Madrid, Spain; Section of Immunology, Universidad San Pablo-CEU, Madrid, Spain. Electronic address: aserrano@h12o.es.
authors.forename: Antonio
authors.initials: A
authors.lastname: Serrano
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D015415:Biomarkers / Q000378:metabolism; D017252:CD3 Complex / Q000276:immunology / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D002478:Cells, Cultured; D016903:Drug Monitoring / Q000379:methods; D005260:Female; D006801:Humans; D007166:Immunosuppressive Agents / Q000009:adverse effects / Q000097:blood / Q000627:therapeutic use*; D016030:Kidney Transplantation / Q000009:adverse effects; D008213:Lymphocyte Activation / Q000187:drug effects; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D008875:Middle Aged; D008934:Mitogens; D015166:Monitoring, Immunologic / Q000379:methods; D010835:Phytohemagglutinins; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D012189:Retrospective Studies; D013997:Time Factors; D016896:Treatment Outcome
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; D017252:CD3 Complex; D007166:Immunosuppressive Agents; D008934:Mitogens; D010835:Phytohemagglutinins; D000255:Adenosine Triphosphate
keywords: ATP levels; ImmuKnow; Immunosuppression; Lymphocyte proliferation; Mixed lymphocyte reaction; WST-1