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82-P: Proficiency testing for ImmuKnow®
Abstracts
S53
82-P
PROFICIENCY TESTING FOR ImmuKnow®. M.R. Carreno,1 P. Ruiz.1,2 1Department of Surgery-Division of Transplantation, The University of Miami Transplant Institute – The Leonard Miller School of Medicine, Miami, FL, USA; 2Department of Pathology, The University of Miami Transplant Institute – The Leonard Miller School of Medicine. Aim: Cell Mediated Immune (CMI) responses from multicenter trials have been always centralized to one institution due to centers variability. While results have been imperative, the clinical use was precluded (turnaround time ⬎4days). Today, CMI can be done and used clinically (⬍24hrs). Nevertheless, an internal “personnel proficiency” (I-PT) and external “laboratory proficiency” (E-PT) for Immuknow must be done. We have enrolled (since 2007) up to eleven centers to participate in a twice/year E-PT and present here evaluations, difficulties, and technical issues. Methods: Specimens from healthy volunteer and immunosuppressed (IS) patients were tested in a blindfolded manner. Samples were tested on the day draw and after 24 hours under different conditions (motions and blood volume/containers). We sent E-PT to other centers including ourselves. All shipping was done with manufacturer’s precautions and results evaluated using quantitative and qualitative FDA guidelines. Results: In I-PT there was consistent reproducibility, in the quantitative but not in the qualitative assessment when specimens were close to zone changes. In E-PT, healthy volunteers showed excellent correlation with in-house results. Shipped specimens from IS patients showed good correlation when compared overall results. In addition, centers variations were reduced by elimination of air apace within sent out containers. Conclusions: We recommend Immuknow users to have an I-PT and E-PT exchanges before attempting to do clinical interpretations. Once proficient, we emphasize the need for pre and post-transplant serial testing in conjunction with a qualitative assessment of the patient dependent values. Immuknow is an additional powerful biomarker tool in the management of transplant patients.
83-P
SERIAL CYLEX- ImmuKnow® TESTING IS BENEFICIAL IN INTESTINAL TRANSPLANTATION. M.R. Carreno, P. Tryphonopoulos, E. Island, L. Smith, A. Tekin, S. Nishida, D. Levi, A. Tzakis, P. Ruiz. DEPTs Surgery & Pathology, Univ. of Miami, Miami, FL. Aim: Human intestinal transplantation (ITx) has technically improved and been adopted as a viable treatment modality for short bowel syndrome. During this time more effective immunosuppression (IS) and new tools to monitor immunity have developed. We present here the potential benefits of serial monitoring cell mediated immunity (CMI) using the ImmuKnow assay. Methods: We evaluated 28 recipients throughout the first year post-ITx. Patients received ab induction and maintained on Prograf and low steroids. The global CMI was studied serially in a quantitative and qualitative manner with Immuknow. Results: Pre-ITx mean responses were 287.9 ⫾162.2 ng/ml, significantly lower than healthy controls (432.8⫾95.3) or ESRD (342.1⫾155) pre-liver (332.8⫾123.5), pre-heart (338.9⫾143.2) or pre-Lung (439.7⫾123.8) Tx. At two months post-ITx a decrease to 271.3⫾152.4 ng/ml was observed while at 12 months post-ITx, a return to baseline was evident (289.6⫾211.2) (n⫽8). The mean IS trough levels were overall higher (9.5 vs 6.3 ng/ml) than in all other solid organs Tx. An inverted peripheral CD4:CD8 ratios were the norm and CD4 enumeration did not correlate with ImmuKnow response. Patients that tested higher pre-Tx remained higher post-ITx without notable evidence of acute rejection and those low followed a similar curve. While the numbers of patients are few to subgroup, we found an inverse correlation of IS with the degree of CMI. Of those with infections, there was a tendency to drop ⬎150 from pre-ITx level. Conclusions: Serial monitoring with ImmuKnow is crucial to establish baselines. Significant changes throughout the post-Tx helps individualize and tailor IS. Our results suggest that Immuknow can aid significantly in the management of post-ITx recipients.
S53
82-P
PROFICIENCY TESTING FOR ImmuKnow®. M.R. Carreno,1 P. Ruiz.1,2 1Department of Surgery-Division of Transplantation, The University of Miami Transplant Institute – The Leonard Miller School of Medicine, Miami, FL, USA; 2Department of Pathology, The University of Miami Transplant Institute – The Leonard Miller School of Medicine. Aim: Cell Mediated Immune (CMI) responses from multicenter trials have been always centralized to one institution due to centers variability. While results have been imperative, the clinical use was precluded (turnaround time ⬎4days). Today, CMI can be done and used clinically (⬍24hrs). Nevertheless, an internal “personnel proficiency” (I-PT) and external “laboratory proficiency” (E-PT) for Immuknow must be done. We have enrolled (since 2007) up to eleven centers to participate in a twice/year E-PT and present here evaluations, difficulties, and technical issues. Methods: Specimens from healthy volunteer and immunosuppressed (IS) patients were tested in a blindfolded manner. Samples were tested on the day draw and after 24 hours under different conditions (motions and blood volume/containers). We sent E-PT to other centers including ourselves. All shipping was done with manufacturer’s precautions and results evaluated using quantitative and qualitative FDA guidelines. Results: In I-PT there was consistent reproducibility, in the quantitative but not in the qualitative assessment when specimens were close to zone changes. In E-PT, healthy volunteers showed excellent correlation with in-house results. Shipped specimens from IS patients showed good correlation when compared overall results. In addition, centers variations were reduced by elimination of air apace within sent out containers. Conclusions: We recommend Immuknow users to have an I-PT and E-PT exchanges before attempting to do clinical interpretations. Once proficient, we emphasize the need for pre and post-transplant serial testing in conjunction with a qualitative assessment of the patient dependent values. Immuknow is an additional powerful biomarker tool in the management of transplant patients.
83-P
SERIAL CYLEX- ImmuKnow® TESTING IS BENEFICIAL IN INTESTINAL TRANSPLANTATION. M.R. Carreno, P. Tryphonopoulos, E. Island, L. Smith, A. Tekin, S. Nishida, D. Levi, A. Tzakis, P. Ruiz. DEPTs Surgery & Pathology, Univ. of Miami, Miami, FL. Aim: Human intestinal transplantation (ITx) has technically improved and been adopted as a viable treatment modality for short bowel syndrome. During this time more effective immunosuppression (IS) and new tools to monitor immunity have developed. We present here the potential benefits of serial monitoring cell mediated immunity (CMI) using the ImmuKnow assay. Methods: We evaluated 28 recipients throughout the first year post-ITx. Patients received ab induction and maintained on Prograf and low steroids. The global CMI was studied serially in a quantitative and qualitative manner with Immuknow. Results: Pre-ITx mean responses were 287.9 ⫾162.2 ng/ml, significantly lower than healthy controls (432.8⫾95.3) or ESRD (342.1⫾155) pre-liver (332.8⫾123.5), pre-heart (338.9⫾143.2) or pre-Lung (439.7⫾123.8) Tx. At two months post-ITx a decrease to 271.3⫾152.4 ng/ml was observed while at 12 months post-ITx, a return to baseline was evident (289.6⫾211.2) (n⫽8). The mean IS trough levels were overall higher (9.5 vs 6.3 ng/ml) than in all other solid organs Tx. An inverted peripheral CD4:CD8 ratios were the norm and CD4 enumeration did not correlate with ImmuKnow response. Patients that tested higher pre-Tx remained higher post-ITx without notable evidence of acute rejection and those low followed a similar curve. While the numbers of patients are few to subgroup, we found an inverse correlation of IS with the degree of CMI. Of those with infections, there was a tendency to drop ⬎150 from pre-ITx level. Conclusions: Serial monitoring with ImmuKnow is crucial to establish baselines. Significant changes throughout the post-Tx helps individualize and tailor IS. Our results suggest that Immuknow can aid significantly in the management of post-ITx recipients.