Role of macroscopic on-site evaluation of endoscopic ultrasoundguided fine-needle aspiration/biopsy:Results of a multicentric prospective study

BACKGROUND The concept of macroscopic on-site evaluation(MOSE)was introduced in 2015 when the endoscopist observed better diagnostic yield when the macroscopically visible core on MOSE was superior to 4 mm.Recent studies suggest that MOSE by the endoscopist may be an excellent alternative to rapid on-site evaluation,and some classi-fications have been published.Few studies have assessed the adequacy of histologic cores in MOSE during endoscopic ultrasound-guided fine-needle aspiration/biopsy(EUS-FNA/FNB).AIM To evaluate the performance of MOSE during EUS-FNA/FNB.METHODS This multicentric prospective study was conducted in 16 centers in 3 countries(Egypt,Iraq,and Morocco)and included 1108 patients with pancreatic,biliary,or gastrointestinal pathology who were referred for EUS examination.We prospectively analyzed the MOSE in 1008 patients with available histopathological reports according to 2 classifications to determine the adequacy of the histological core samples.Data management and analysis were performed using a Statistical Package for Social Sciences(SPSS)version 27.RESULTS A total of 1074 solid lesions were biopsied in 1008 patients with available cytopathological reports.Mean age was 59 years,and 509 patients(50.5%)were male.The mean lesion size was 38 mm.The most frequently utilized needles were FNB-Franseen(74.5%)and 22 G(93.4%),with a median of 2 passes.According to 2 classifications,618 non-bloody cores(61.3%)and 964 good samples(95.6%)were adequate for histological evaluation.The overall diagnostic yield of cytopathology was 95.5%.The cytological examination confirmed the diagnosis of malignancy in 861 patients(85.4%),while 45 samples(4.5%)were inconclusive.Post-procedural adverse events occurred in 33 patients(3.3%).Statistical analysis showed a difference between needle types(P=0.035)with a high sensitivity of FNB(97%).The analysis of the relationship between the MOSE-score and the final diagnosis showed a significant difference between the different scores of the MOSE(P<0.001).CONCLUSION MOSE is a simple method that allows endoscopists to increase needle passes to improve sample quality.There is significantly higher FNB sensitivity and cytopathology diagnostic yield with good MOSE cores.

Systemic air embolism after transthoracic lung biopsy: A case report and review of literature

Computed tomography (CT)-guided lung biopsy is a common diagnostic procedure that is associated with various complications, including pneumothorax, hemoptysis and parenchymal hemorrhage. Systemic air embolism is a very rare (0.07%) but potentially lifethreatening complication. We report a fatal case of air embolism to the cerebral and coronary arteries confirmed by head and chest CT, followed by a review of the literature.

Fine-needle aspiration technique under endoscopic ultrasound guidance:A technical approach for RNA profiling of pancreatic neoplasms

BACKGROUND Early diagnosis of pancreatic ductal adenocarcinoma(PDAC)has been a longstanding challenge.The prognosis of patients with PDAC depends on the stage at diagnosis.It is necessary to identify biomarkers for the detection and differentiation of pancreatic tumors and optimize PDAC sample preparation procedures for DNA and RNA analysis.Most molecular studies are done using paraffin-embedded blocks;however,the integrity of DNA and RNA is often compromised in this format.Moreover,RNA isolated from human pancreatic tissue samples is generally of low quality,in part,because of the high concentration of endogenous pancreatic RNAse activity present.AIM To assess the potential of endoscopic ultrasound-guided fine-needle aspiration(EUS-FNA)to obtain specimens from pancreatic neoplasms for subsequent RNA molecular profiling,including next-generation sequencing(NGS).METHODS Thirty-four EUS-FNA samples were included in this study:PDAC(n=15),chronic pancreatitis(n=5),pancreatic cysts(n=14),mucinous cysts(mucinous cystic neoplasia/intraductal papillary mucinous neoplasia)n=7,serous cystic neoplasms n=5,and pseudocysts n=2.Cyst material consisted of cyst fluid and cyst wall samples obtained by through-the-needle biopsy(TTNB).Samples were stored at -80℃ until analysis.RNA purity(A260/230,A260/280 ratios),concentration,and integrity(RIN)were assessed.Real-time polymerase chain reaction was conducted on all samples,and small RNA libraries were prepared from solid mass samples.RESULTS RNA was successfully extracted from 29/34(85%)EUS-FNA samples:100% pancreatic adenocarcinoma samples,100% chronic pancreatitis samples,70% pancreatic fluid cyst samples,and 50%TTNB samples.The relative expression of GAPDH and HPRT were obtained for all successfully extracted RNA samples(n=29)including lowquality RNA specimens.Low concentration and nonoptimal RIN values(no less than 3)of RNA extracted from EUS-FNA samples did not prevent NGS library preparation.The suitability of cyst fluid samples for RNA profiling varied.The quality of RNA extracted from mucinous cyst fluid had a median RIN of 7.7(5.0-8.2),which was compatible with that from solid neoplasms[6.2(0-7.8)],whereas the quality of the RNA extracted from all fluids of serous cystic neoplasms and TTNB samples had a RIN of 0.CONCLUSION The results demonstrate the high potential of EUS-FNA material for RNA profiling of various pancreatic lesions,including low-quality RNA specimens.

Endoscopic ultrasound-guided fine-needle aspiration biopsy-Recent topics and technical tips

Endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNA) is a useful procedure that enables reliable pathological diagnoses of pancreatobiliary diseases, subepithelial lesions, and swollen lymph nodes. In recent years, a pathological diagnosis based on EUS-FNA has made it possible to provide accurate treatment methods not only in these fields, but also in respiratory organs and otorhinolaryngology. This review discusses the latest topics pertaining to EUS-FNA as well as procedural tips.

Cheesy material on macroscopic on-site evaluation after endoscopic ultrasound-guided fine-needle biopsy:Don't miss the tuberculosis

Endoscopic ultrasound-guided fine-needle biopsy(EUS-FNB)is an excellent investigation to diagnose pancreatic lesions and has shown high accuracy for its use in pathologic diagnosis.Recently,macroscopic on-site evaluation(MOSE)performed by an endoscopist was introduced as an alternative to rapid on-site cytologic evaluation to increase the diagnostic yield of EUS-FNB.The MOSE of the biopsy can estimate the adequacy of the sample directly by the macroscopic evaluation of the core tissue obtained from EUS-FNB.Isolated pancreatic tuberculosis is extremely rare and difficult to diagnose because of its non-specific signs and symptoms.Therefore,this challenging diagnosis is based on endoscopy,imaging,and the bacteriological and histological examination of tissue biopsies.This uncommon presentation of tuberculosis can be revealed as pancreatic mass mimicking cancer.EUS-FNB can be very useful in providing a valuable histopathological diagnosis.A calcified lesion with a cheesy core in MOSE must be suggestive of tuberculosis,leading to the request of the GeneXpert,which can detect Mycobacterium tuberculosis deoxyribonucleic acid and resistance to rifampicin.A decent diagnostic strategy is crucial to prevent unnecessary surgical resection and to supply conservative management with antitubercular therapy.

Needle tract seeding of papillary thyroid carcinoma after fine-needle capillary biopsy:A case report

BACKGROUND Fine-needle biopsy is an accurate and cost-efficient tool for the assessment of thyroid nodules.It includes two primary methods:Fine-needle capillary biopsy(FNCB)and fine-needle aspiration biopsy.Needle tract seeding(NTS)is a rare complication of thyroid fine-needle biopsy mainly caused by fine-needle aspiration biopsy rather than FNCB.Here,we present an extremely rare case of a papillary thyroid carcinoma(PTC)patient with FNCB-derived NTS.CASE SUMMARY We report a 32-year-old woman with PTC who showed subcutaneous NTS 1 year after FNCB and thyroidectomy.NTS was diagnosed based on clinical manifestations,biochemistry indices,and imaging(computed tomography and ultrasound).Pathological identification of PTC metastases consistent with the puncture path is the gold standard for diagnosis.Surgical resection was the main method used to treat the disease.After surgery,thyroid function tests and ultrasound scans were performed every 3-6 mo.To date,no evidence of tumor recurrence has been observed.CONCLUSION FNCB is a safe procedure as NTS is rare,and can be easily removed surgically with no recurrence.Accordingly,NTS should not limit the usefulness of FNCB.

Sub-acute disseminated pulmonary tuberculosis in a girl diagnosed by CT guided transthoracic needle biopsy: case report and literature review

Primary tuberculosis often occurs in children with the main types of hilar lymphonode tuberculosis or Ghon complex. When pulmonary tuberculosis aggravates and tubercule bacillus invades pulmonary veins, a disseminated disease may occur clinically. One 8-year-old girl presented with hypodynamia and emaciation. CT indicated the disseminated disease and extensive mediastinal and hilar lymphadenopathy. But she had negative sputum smears and negative PPD test. The bronchi showed inflammatory change in fiberoptic bronchoscopy. Percutaneous transthoracic needle biopsy (PTNB) guided by CT was used as a means for the diagnosis of pulmonary tuberculosis. The case was rapidly diagnosed as sub-acute disseminated pulmonary tuberculosis with the help of combined CT imagine and histology.

The efficacy and safety of endoscopic ultrasound-guided fine-needle biopsy in gallbladder masses

Background: Endoscopic ultrasound-guided fine-needle biopsy(EUS-FNB) is a widely used modality for acquiring various target samples, but its efficacy in gallbladder masses is unknown. The aim of this retrospective study was to evaluate the efficacy and safety of EUS-FNB in patients with gallbladder masses. Methods: The study samples were composed of patients from March 2015 to July 2019 who needed to identify the nature of gallbladder masses through EUS-FNB. The outcomes of this study were the adequacy of specimens, diagnostic yields, technical feasibility, and adverse events of the EUS-FNB in gallbladder masses. Results: A total of 27 consecutive patients with a median age of 58 years were included in this study. The 22-gauge FNB needle was feasible in all lesions. The median follow-up period of the patients was 294 days. The specimens sufficient for diagnosis account for 89%(24/27) and 93%(25/27) in cytology and histology, respectively. The overall diagnostic yields for malignancy showed the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 95.45% [95% confidence interval(CI): 75.12%-99.76%], 100%(95% CI: 46.29%-10 0%), 10 0%(95% CI: 80.76%-100%), 83.33%(95% CI: 36.48%-99.12%), and 96.30%(95% CI: 80.20%-99.99%), respectively. The subgroup analysis revealed that FNB could obtain sufficient specimens and high diagnostic yields in both gallbladder mass < 20.5 mm group and ≥20.5 mm group. One patient experienced mild abdominal pain after the procedure and recovered within one day. Conclusions: EUS-FNB is a reasonable diagnostic tool for the pretreatment diagnosis of patients with gallbladder masses, especially for patients who may miss the opportunity of surgery and need sufficient specimens to identify the pathological type so as to determine chemotherapy regimens. Further largescale studies are needed to confirm our conclusion.

Clinical value of CTS-guided percutaneous transthoracic biopsy in elder patients with lung cancer

Objective： To evaluate the value of CTS-guided percutaneous transthoracic biopsy for the diagnosis of pulmonary lesions in elder patients. Methods： The intact data of 78 elder patients underwent percutaneous pulmonary biopsy with a spring-core biopsy needle under CTS guidance were analyzed. The diagnosis of all the cases was confirmed by pathology and follow-up. Results： Ninety-five lung biopsies were performed in 78 cases and satisfactory tissue specimens obtained in all cases, the success rate was 100%. The diagnosis for malignant tumors and benign lesions was 72 and 6, respectively. The overall accuracy rate was 100%. Pneumothorax was noted in 5 cases （6.4%） and hemoptysis seen in 10 patients （12.8%）. Conclusion： CTS-guided percutaneous transthoracic biopsy with a spring-core biopsy needle are accurate, safe and quick for pulmonary lesions in elder patients.

Diagnosis of hepatic inflammatory pseudotumor by fine-needle biopsy

Hepatic inflammatory pseudotumor(IPT)is a benign lesion characterized by chronic infiltration of inflammatory cells and fibrosis that clinically,radiologically,and pathologically mimics malignancy.However,the epidemiology of IPTs remains unclear.IPTs are often misdiagnosed as malignant lesions because of the lack of characteristic features.We present the case of a 32-year-old man that was misdiagnosed as intrahepatic cholangiocarcinoma by CECT,CEMRI,and CEUS,which was finally confirmed as IPT by fine-needle liver biopsy.In this report,the key factor in the diagnosis of liver inflammatory masses was the presence of hepatic perfusion disorder.

Influence of the safety and diagnostic accuracy of preoperative endoscopic ultrasound-guided fine-needle aspiration for resectable pancreatic cancer on clinical performance

AIM: To evaluate the safety and diagnostic accuracy of endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) in a cohort of pancreatic cancer patients.

Role of endoscopic ultrasound fine-needle aspiration evaluating adrenal gland enlargement or mass

AIM： To report the clinical impact of adrenal endoscop-ic ultrasound fine-needle aspiration （EUS-FNA） in the evaluation of patients with adrenal gland enlargement or mass.METHODS： In a retrospective single-center case-series, patients undergoing EUS-FNA of either adrenal gland from 1997-2011 in our tertiary care center were included. Medical records were reviewed and results of EUS, cytology, adrenal size change on follow-up imag-ing ≥ 6 mo after EUS and any repeat EUS or surgery were abstracted. A lesion was considered benign if： （1） EUS-FNA cytology was benign and the lesion remained 〈 1 cm from its original size on follow-up computed tomography （CT）, magnetic resonance imaging or repeat EUS ≥ 6 mo after EUS-FNA; or （2） subsequent adrenalectomy and surgical pathology was benign. RESULTS： Ninety-four patients had left （n = 90） and/or right （n = 5） adrenal EUS-FNA without adverse events. EUS indications included： cancer staging or suspected recurrence （n = 31）, pancreatic （n = 20）, medi-astinal （n = 10）, adrenal （n = 7）, lung （n = 7） mass or other indication （n = 19）. Diagnoses after adrenal EUS-FNA included metastatic lung （n = 10）, esophageal （n= 5）, colon （n = 2）, or other cancer （n = 8）; benign primary adrenal mass or benign tissue （n = 60）; or was non-diagnostic （n = 9）. Available follow-up confrmed a benign lesion in 5/9 non-diagnostic aspirates and 32/60 benign aspirates. Four of the 60 benign aspirates were later confrmed as malignant by repeat biopsy, follow-up CT, or adrenalectomy. Adrenal EUS-FNA diagnosed metastatic cancer in 24, and ruled out metastasis in 10 patients. For the diagnosis of malignancy, EUS-FNA of either adrenal had sensitivity, specifcity, positive predic-tive value and negative predictive value of 86%, 97%, 96% and 89%, respectively.CONCLUSION： Adrenal gland EUS-FNA is safe, mini-mally invasive and a sensitive technique with signifcant impact in the management of adrenal gland mass or enlargement.

Core needle biopsy for thyroid nodules assessment-a new horizon?

Ultrasound-guided fine-needle aspiration is the standard for evaluating thyroid nodules with a high safety profile and a relatively low number of non-diagnostic cytological findings.Nevertheless,this diagnostic method traditionally has its weak points.Several diagnostic categories such as BethesdaⅠ,ⅢandⅣare not reliable for thyroid carcinoma risk assessment.Recent advancements in a core needle biopsy made it possible to use this tool as a new method for thyroid nodules evaluation.The main feature of this method is the use of thin needles(18-21G)and guns with an automatic trigger mechanism.The histological material collected with the use of a core needle biopsy is usually superior to cytological.Therefore,the core needle biopsy can be used as a complementary technique to a standard fine needle aspiration in difficult and dubious cases of thyroid neoplasia with uncertain malignant potential.

Stage IV malignant transformation of mature cystic teratoma palliatively treated with concurrent chemoradiotherapy:A case report

BACKGROUND Malignant transformation(MT)of mature cystic teratoma(MCT)has a poor prognosis,especially in advanced cases.Concurrent chemoradiotherapy(CCRT)has an inhibitory effect on MT.CASE SUMMARY Herein,we present a case in which CCRT had a reduction effect preoperatively.A 73-year-old woman with pyelonephritis was referred to our hospital.Computed tomography revealed right hydronephrosis and a 6-cm pelvic mass.Endoscopic ultrasound-guided fine-needle biopsy(EUS-FNB)revealed squamous cell carci-noma.The patient was diagnosed with MT of MCT.Due to her poor general con-dition and renal malfunction,we selected CCRT,expecting fewer adverse effects.After CCRT,her performance status improved,and the tumor size was reduced;surgery was performed.Five months postoperatively,the patient developed dis-semination and lymph node metastases.Palliative chemotherapy was ineffective.She died 18 months after treatment initiation.CONCLUSION EUS-FNB was useful in the diagnosis of MT of MCT;CCRT suppressed the disea-se and improved quality of life.

Mucosal-incision assisted biopsy for suspected gastric gastrointestinal stromal tumors

To evaluate the diagnostic yield of the procedure, mucosal-incision assisted biopsy (MIAB), for the histological diagnosis of gastric gastrointestinal stromal tumor (GIST), we performed a retrospective review of the 27 patients with suspected gastric GIST who underwent MIAB in our hospitals. Tissue samples obtained by MIAB were sufficient to make a histological diagnosis (diagnostic MIAB) in 23 out of the 27 patients, where the lesions had intraluminal growth patterns. Alternatively, the samples were insufficient (non-diagnosticMIAB) in remaining 4 patients, three of whom had gastric submucosal tumor with extraluminal growth patterns. Although endoscopic ultrasound and fine needle aspiration is the gold standard for obtaining tissue specimens for histological and cytological analysis of suspected gastric GISTs, MIAB can be used as an alternative method for obtaining biopsy specimens of lesions with an intraluminal growth pattern.

Endoscopic ultrasound-guided through-the-needle microforceps biopsy and needle-based confocal laser-endomicroscopy increase detection of potentially malignant pancreatic cystic lesions:A single-center study

BACKGROUND Currently,there is insufficient data about the accuracy in the diagnosing of pancreatic cystic lesions(PCLs),especially with novel endoscopic techniques such as with direct intracystic micro-forceps biopsy(mFB)and needle-based confocal laser-endomicroscopy(nCLE).AIM To compare the accuracy of endoscopic ultrasound(EUS)and associated techniques for the detection of potentially malignant PCLs:EUS-guided fine needle aspiration(EUS-FNA),contrast-enhanced EUS(CE-EUS),EUS-guided fiberoptic probe cystoscopy(cystoscopy),mFB,and nCLE.METHODS This was a single-center,retrospective study.We identified patients who had undergone EUS,with or without additional diagnostic techniques,and had been diagnosed with PCLs.We determined agreement among malignancy after 24-mo follow-up findings with detection of potentially malignant PCLs via the EUSguided techniques and/or EUS-guided biopsy when available(EUS malignancy detection).RESULTS A total of 129 patients were included, with EUS performed alone in 47/129. In 82/129 patients,EUS procedures were performed with additional EUS-FNA (21/82), CE-EUS (20/82), cystoscopy(27/82), mFB (36/82), nCLE (44/82). Agreement between EUS malignancy detection and the 24-mo follow-up findings was higher when associated with additional diagnostic techniques thanEUS alone [62/82 (75.6%) vs 8/47 (17%);OR 4.35, 95%CI: 2.70-7.37;P < 0.001]. The highestmalignancy detection accuracy was reached when nCLE and direct intracystic mFB were bothperformed, with a sensitivity, specificity, positive predictive value, negative predictive value andobserved agreement of 100%, 89.4%, 77.8%, 100% and 92.3%, respectively (P < 0.001 comparedwith EUS-alone).CONCLUSIONThe combined use of EUS-guided mFB and nCLE improves detection of potentially malignantPCLs compared with EUS-alone, EUS-FNA, CE-EUS or cystoscopy.

Endoscopic ultrasound guided liver biopsy: Recent evidence

Liver biopsy(LB)is an essential tool in diagnosing,evaluating and managing various diseases of the liver.As such,histopathological results are critical as they establish or aid in diagnosis,provide information on prognosis,and guide the appropriate selection of medical therapy for patients.Indications for LB include evaluation of persistent elevation of liver chemistries of unclear etiology,diagnosis of chronic liver diseases such as Wilson's disease,autoimmune hepatitis,small duct primary sclerosing cholangitis,work up of fever of unknown origin,amyloidosis and more.Traditionally,methods of acquiring liver tissue have included percutaneous LB(PCLB),transjugular LB(TJLB)or biopsy taken surgically via laparotomy or laparoscopy.However,traditional methods of LB may be inferior to newer methods.Additionally,PCLB and TJLB carry higher risks of adverse events and complications.More recently,endoscopic ultrasound guided LB(EUS-LB)has evolved as an alternative method of tissue sampling that has proven to be safe and effective,with limited adverse events.Compared to PC and TJ routes,EUS-LB may also have a greater diagnostic yield of tissue,be superior for a targeted approach of focal lesions,provide higher quality images and allow for greater patient comfort.These advantages have contributed to the increased use of EUS-LB as a technique for obtaining liver tissue.Herein,we provide a review of the recent evidence of EUS-LB for liver disease.

Characterizing gastrointestinal stromal tumors and evaluating neoadjuvant imatinib by sequencing of endoscopic ultrasound-biopsies

AIM To evaluate endoscopic ultrasound(EUS)-guided biopsies for the pretreatment characterization of gastrointestinal stromal tumors(GIST) to personalize the management of patients.METHODS All patients with lesions suspected to be GIST who were referred for EUS-sampling at a tertiary Swedish center were eligible for inclusion 2006-2015. During the observational study phase(2006-2011), routine fine-needle-aspiration(EUS-FNA) was performed.In 2012-2015, we converted to an interventional, randomized protocol with dual sampling EUS-FNA and fine-needle-biopsy-sampling(EUS-FNB) for all lesions. c-KIT-and DOG-1-immunostaining was attempted in all samples and a manual count of the Ki-67-index was performed. FNB-sampled tissue and the resected specimens were subjected to Sanger sequencing of the KIT and platelet-derived growth factor alpha(PDGFRA) genes. RESULTS In all, 64 unique patients with GIST were included, and of these, 38 were subjected to pretreatment dual sampling. EUS-FNB had a higher diagnostic sensitivity when compared head-to-head with EUS-FNA(98% vs 58%, P < 0.001) and was more adequate for Ki-67-indexing(Ki-67EUS)(92% vs 40%, P < 0.001). Sequencing of EUS-biopsies was successful in 43/44(98%) patients, and the mutation profiles(KIT-mutation 73%, PDGFRA-mutation 18%, wild-type 7%) were fully congruent with those detected in the corresponding resected specimens. In imatinib-na?ve patients, the Ki-67_(EUS) was comparable with the Ki-67-index in the corresponding surgical specimens(Ki-67_(SURG))(2.7% vs 2.9%, P = 0.68). In patients treated with neoadjuvant imatinib who also carried mutations indicating sensitivity, the Ki-67 EUS was higher than the Ki-67_(SURG)(2.5% vs 0.2%, P = 0.005), with a significant reduction in the Ki-67-index of-91.5%(95%CI:-82.4 to-96.0, P = 0.005). CONCLUSION EUS-guided biopsy sampling is accurate for the pretreatment diagnosis and characterization of GISTs and allows the prediction and evaluation of tumor response to neoadjuvant imatinib therapy.

Co-Inhibitors of Second Signal of Lymphocyte Response in Human Renal Transplants: PD-L2, GITR, and ILT-2/3/5 Positive Cells from Aspiration Biopsies Associate with Acute Rejection-Freedom

Following organ transplantation, the outcome of the encounter between an APC and a T lymphocyte is strongly dependent on the presence of costimulatory and co-inhibitory molecules, the former associated with allograft rejection and the latter with allograft acceptance. We evaluated the expression of PD-L2, GITR, ILT-2/3/5, and ILT-4 on graft-infiltrating cells procured by Fnab from human KTx under different immunosuppressive regimens. Methods: Fnab biopsies were performed on days 7 or 14 - 30 in stable KTx and on the day of acute rejection diagnosis. Cytopreparations were studied by the enzymatic avidin biotin complex staining. Results: Acute rejection group showed a significant down-regulated expression of PD-L2, GITR, and ILT-2/3/5 as compared to stable group, while for ILT-4 we did not find significant difference. Anti-IL2αR and rapamicyn treatment trend to down-regulate ILT-4 expression, although meaningless. A significantly positive correlation was observed between PD-L2 and GITR expression in Fnab. The PPV for acute rejection diagnosis for both PD-L2 and GITR was clearly above 0.8. Conclusions: Our findings point to an early entrance of cells expressing PD-L2, GITR and ILT-2/3/5 inside human KTx who are going to remain rejection-free. Both PD-L2 and GITR shared a high ability to rule-in and rule-out acute rejection.