Post-thaw Viability of Cryopreserved Hematopoietic Progenitor Cell Grafts: Does It Matter?

Cell viability in peripheral blood progenitor cell (PBPC) grafts and its influence on the clinical course following transplantation was evaluated in 81 consecutive transplantations (72 autologous, 9 allogeneic) performed in patients with hematological diseases. Viability of cells in PBPC grafts immediately upon collection was 98.6±3.5%, after addition of dimethyl sulfoxide (DMSO) 73.3±21.8%, and post-thaw 65.2±16.1%. It did not differ significantly between patients with different diagnoses, gender, age, type of priming used, dose of G-CSF administered or number of CD34+ cells collected. However, grafts stored for more than 60 days showed lower post-thaw viability compared to the ones thawed in the 60 days following cryopreservation (56.6±15.2% vs. 67.6±15.5%, p=0.04). Post-thaw graft viability did not influence engraftment time, but there was a predisposition towards infectious complications in the post-transplant period in patients receiving grafts with lower percentage of viable cells. They developed febrile neutropenia more often (72.2% vs. 50% of patients, p=0.05) and had more febrile days (2.4±2.6 vs. 1.5±2.3, p=0.05) following transplantation. We have demonstrated that PBPC grafts are capable of long term engraftment regardless of the graft storage time or percentage of viable cells post-thaw, which confirms the robustness of CD34+ cells during the freeze/thaw procedures carried out in daily clinical practice. Granulocyte concentration in PBPC grafts could have an influence on infectious complications following transplantation and needs to be further investigated on a larger number of patients

An Unusual Presentation of Gaucher’s Disease: Aortic Valve Fibrosis in a Patient Homozygous for a Rare G377S Mutation

Gaucher’s disease (GD) has variable presentations, but cardiac involvement is a generally uncommon clinical manifestation of the disease. In the past 25 years, the underlying genetic disorder in GD has been well characterized, with almost 300 mutations identified in the glucocerebrosidase gene (GBA). Nevertheless, clear genotype-phenotype correlations have been confirmed only for the most frequent mutations. We present a female patient, who was known to have aortic valve pathology from the age of 30. Despite medical follow up, at the age of 60 she presented with heart failure (NYHA III). At that time echocardiography showed severe fibrosed aortic valve stenosis. Valvuloplasty was planned, when thrombocytopenia, previously considered to be autoimmune, became severe. Anemia and leukopenia were also noted. Moderate splenomegaly and severe bone marrow infiltration were found on MRI. Bone marrow aspiration revealed typical Gaucher cells and the enzyme activity assay confirmed the diagnosis. DNA investigation showed that the patient is homozygous for the G377S mutation. To our knowledge, of all mutations identified so far, only homozygosity for the D409H mutation has been associated with cardiovascular valvular disease in patients with a rare type 3c GD. G377S, found in our patient, is a rare mutation, previously reported as a \u27mild’ mutation, because of the finding that homoallelic patients were essentialy asymptomatic or had mild disease. Our patient, also homozygous for G377S mutation, had a severe form of type 1 GD, with rare cardiac valve involvement, which is a previously unreported clinical presentation for this mutation. This case further proves that patients with the same genotypes can have different phenotypes, emphasizing the influence of other genetic and/or environmental factors

Post-thaw Viability of Cryopreserved Hematopoietic Progenitor Cell Grafts: Does It Matter?

Cell viability in peripheral blood progenitor cell (PBPC) grafts and its influence on the clinical course following transplantation was evaluated in 81 consecutive transplantations (72 autologous, 9 allogeneic) performed in patients with hematological diseases. Viability of cells in PBPC grafts immediately upon collection was 98.6±3.5%, after addition of dimethyl sulfoxide (DMSO) 73.3±21.8%, and post-thaw 65.2±16.1%. It did not differ significantly between patients with different diagnoses, gender, age, type of priming used, dose of G-CSF administered or number of CD34+ cells collected. However, grafts stored for more than 60 days showed lower post-thaw viability compared to the ones thawed in the 60 days following cryopreservation (56.6±15.2% vs. 67.6±15.5%, p=0.04). Post-thaw graft viability did not influence engraftment time, but there was a predisposition towards infectious complications in the post-transplant period in patients receiving grafts with lower percentage of viable cells. They developed febrile neutropenia more often (72.2% vs. 50% of patients, p=0.05) and had more febrile days (2.4±2.6 vs. 1.5±2.3, p=0.05) following transplantation. We have demonstrated that PBPC grafts are capable of long term engraftment regardless of the graft storage time or percentage of viable cells post-thaw, which confirms the robustness of CD34+ cells during the freeze/thaw procedures carried out in daily clinical practice. Granulocyte concentration in PBPC grafts could have an influence on infectious complications following transplantation and needs to be further investigated on a larger number of patients

Amniotic membrane transplantation for severe ocular graft-versus-host disease following allogeneic hematopoietic stem cell transplantation

Allogeneic stem cell transplantation (allo-SCT) offers cure to otherwise incurable hematologic malignancies, but can also lead to many infectious and immune complications, most importantly graft-versus-host disease (GVHD). Ocular GVHD (oGVHD) occurs in 40-60% of allo-SCT patients and can result in severe ocular surface disease causing vision impairment and deterioration of quality of life. Amniotic membrane transplantation (AMT) is an established technique in the treatment of various diseases of the ocular surface. This method could provide new options in the management of otherwise disabling severe oGVHD We describe a young female patient with myeloproliferative neoplasm who underwent allo-SCT from an unrelated donor and suffered from numerous post-transplant complications. In the early post-transplant period she developed acute skin and liver GVHD, requiring introduction of immunosupressive treatment with steroids. Steroid treatment was then complicated with miopathy, iatrogenic diabetes and many infections. She developed serious herpes virus (HSV) ophtalmitis followed by severe GVHD of the eye. Despite multiagent local therapy, oGVHD progressed to ocular ulcers with threatening corneal perforation. We hesitated from increasing systemic immunosupression due to severity of previous HSV reactivation. Therefore we decided to perform AMT which led to complete corneal healing and full clinical recovery. Moreover, there was no recurrence of severe oGVHD and the patient resumed her daily activities In conclusion, this case report serves as a foundation for further research of AMT possibilities. This procedure could become beneficial in the treatment of severe oGVHD, especially in patients at high risk for infectious complications and contraindication for systemic immunosuppression

Diffuse Large B-cell Lymphoma in Patient after Treatment of angioimmunoblastic T-cell Lymphoma

Relatively few cases of Epstein-Barr (EBV)-positive B-cell lymphomas arising in patients with angioimmunoblastic T-cell lymphoma (AITL) have been reported. We report a case of AITL in which diffuse large B-cell lymphoma arose 13 months after the initial diagnosis of AITL. In a 36-year-old female patient, evaluated for moderate leukocytosis, peripheral and abdominal lymphadenopathy AITL was diagnosed in March 2008, based on results of fine-needle aspiration cytology (FNAC) of the enlarged cervical and supraclavicular lymph nodes.The diagnosis was also confirmed by immunophenotyping and histopathology of the cervical lymph nodes. The patient initially recieved FED chemotherapy (fludarabine, cyclophosphamide, dexamethasone) followed by elective autologous hematopoietic stem cell transplantation. In April 2009 the patient was hospitalized because of fever, pancytopenia, hyperbilirubinemia and peripheral lymphadenopathy. The FNAC of the enlarged cervical lymph nodes was performed again, but this time the smears were composed of polymorphous population of lymphocytes with the predomination of large cells, CD20+ on immunocytochemical stains. The immunophenotyping confirmed a predomination of monoclonal mature B-cells. Patient had high number of EBV DNA copies in plasma and serologic testing revealed increased titers of EBV VCA IgG and EBV EBNA IgG. CHOP-R chemotherapy (cyclophosphamide, doxorubicin, vincristine, prednisone and rituximab) was then administered, resulting in good partial response of the disease. Reduced intensity allogeneic stem cell transplantation performed thereafter, resulted in complete remission of the disease. AITL is a rare lymphoproliferative disorder in which the neoplastic T-cells represent the minority of the lymph node cell population and almost all cases harbor EBV-infected B-cells. Various authors postulated that immunodeficiency in AITL patients together with immunosupresive effects of cytotoxic drugs, may be responsible for EBV-induced proliferation of latently or newely EBV-infected B-cells with eventual clonal selection and progression to aggressive B-cell lymphoma

Febrilne reakcije nakon infuzije krvotvornih matičnih stanica češće su ukoliko se ne primijeni premedikacija kortikosteroidima što rezultira većom upotrebom antibiotika rano nakon transplantacije

Background: There is no consensus as to the need for steroid premedication before fresh product hematopoietic stem cell (HSC) infusion. In case of febrile reaction post-HSC infusion, on-call staff frequently prescribe antibiotics empirically. Considering the recent data on the value of microbiota and its influence on GVHD incidence, we analysed the frequency of febrile reactions and the use of antibiotic after HSC infusion in 149 consecutive patients. Methods: In the time period between 1/2018 and 10/2019, 149 patients were subject to transplantation in our institution. Per institutional standard operating procedure (SOP), all the patients received premedication before hematopoietic stem cell infusion consisting of 20 mg chloropyramine-chlorid iv, and in case of ABO incompatible graft 1 mg/kg methylprednisolone iv. Retrospective data was collected by using patient charts. Survival probability was calculated by applying Kaplan-Meier method. Results: Fifty-two patients received steroids, 12 patients (23%) developing fever after graft infusion, 46 patients received no steroids, 26 of them (57%) developed fever (p<0.001). There was no difference in the number of patients having positive blood cultures. Nine (17%) and 16 (35%) patients received IV antibiotics in the “steroid” and no-steroid” group, respectively (p<0.05). There was no difference in survival between “steroid” and “no-steroid” group. Conclusions: Even with no difference in the frequency of febrile episodes caused by systemic infection, a significantly more patients not receiving steroid premedication develop fever and are treated with IV antibiotics, which could potentially have further implications on transplantation outcomes due to its influence on microbiota early post-transplant.Uvod: Trenutno ne postoji konsenzus o potrebi primjene premedikacije kortikosteroidima prije infuzije svježih krvotvornih matičnih stanica (KMS). U slučaju febrilne reakcije nakon infuzije KMS-a, dežurno osoblje često propiše antibiotike empirijski. Uzimajući u obzir nedavne podatke o vrijednosti mikrobiote i njezinom utjecaju na incidenciju GVHD-a, analizirali smo učestalost febrilnih reakcija i uporabu antibiotika nakon infuzije KMS-a u 149 uzastopna bolesnika. Metode: U razdoblju između 1/2018. i 10/2019. u našoj je ustanovi transplantirano ukupno 149 pacijenata. Prema institucionalnom standardnom operativnom postupku (SOP), svi su pacijenti primali premedikaciju prije infuzije krvotvornih matičnih stanica koja se sastojala od 20 mg kloropiramin-klorida iv, a u slučaju ABO inkompatibilnog presatka i 1 mg / kg metilprednizolona iv. Retrospektivno su prikupljeni podaci koristeći povijesti bolesti pacijenata. Vjerojatnost preživljavanja izračunata je Kaplan-Meierovom metodom. Rezultati: Pedeset i dva bolesnika su primila kortikosteroide, od njih je 12 bolesnika (23%) razvilo vrućicu nakon infuzije presatka, dok 46 bolesnika nije primilo kortikosteroide, a od njih je 26 (57%) razvilo vrućicu (p <0,001). Nije bilo razlike u broju bolesnika koji su imali pozitivne hemokulture. U skupini koja je primila kortikosteroide, 9 (17%) bolesnika je liječeno iv antibioticima, dok je u skupini koja nije primala kortikosteroide, 16 (35%) bolesnika liječeno iv antibioticima (p <0,05). Nije bilo razlike u preživljenju između te dvije skupine. Zaključci: Čak i bez razlike u učestalosti febrilnih epizoda uzrokovanih sistemskom infekcijom, znatno više pacijenata koji nisu dobili premedikaciju je razvilo vrućicu i liječilo se iv antibioticima, što bi potencijalno moglo imati daljnje implikacije na ishode transplantacije zbog utjecaja na mikrobiotu rano nakon transplantacije

Glycoprotein YKL-40: a novel biomarker of chronic graftvs- host disease activity and severity?

Aim To investigate whether increased YKL-40 levels positively correlate with graft-vs-host disease (cGVHD) activity and severity and if YKL-40 could serve as a disease biomarker. Methods This case-control study was conducted at the University Hospital Centre Zagreb from July 2013 to October 2015. 56 patients treated with hematopoietic stem cell transplantation (HSCT) were included: 35 patients with cGVHD and 21 without cGVHD. There was no difference between groups in age, sex, median time from transplant to study enrollment, intensity of conditioning, type of donor, or source of stem cells. Blood samples were collected at study enrollment and YKL-40 levels were measured with ELISA. Disease activity was estimated using Clinician’s Impression of Activity and Intensity of Immunosuppression scales and disease severity using Global National Institutes of Health (NIH) score. Results YKL-40 levels were significantly higher in cGVHD patients than in controls (P = 0.003). The difference remained significant when patients with myelofibrosis were excluded from the analysis (P = 0.017). YKL-40 level significantly positively correlated with disease severity (P < 0.001; correlation coefficient 0.455), and activity estimated using Clinician’s Impression of Activity (P = 0.016; correlation coefficient 0.412) but not using Intensity of Immunosuppression (P = 0.085; correlation coefficient 0.296). Conclusion YKL-40 could be considered a biomarker of cGVHD severity and activity. However, validation in a larger group of patients is warranted, as well as longitudinal testing of YKL-40 levels in patients at risk of developing cGVHD

BLOODSTREAM INFECTIONS AFTER LIVER AND HEMATOPOIETIC STEM CELL TRANSPLANTATION

Cilj ovog retrospektivnog istraživanja je ustanoviti i usporediti incidenciju, vrijeme pojavnosti kao i etiologiju infekcija krvotoka u bolesnika liječenih ortotopnom transplantacijom jetre (OTJ) ili transplantacijom krvotvornih matičnih stanica (TKMS), na temelju iskustva jedne ustanove. Analizirano je 280 uzastopnih transplantacija u vremenskom razdoblju od 34 mjeseca. Zabilježene su 84 epizode infekcija krvotoka (47 kod OTJ bolesnika, 37 kod TKMS bolesnika) s medijanom nastupa 28 dana nakon transplantacije. Relativna incidencija infekcija krvotoka iznosila je 34,6 (OTJ) i 29,4 (TKMS) epizoda na 100 bolesnika, te nije uočena statistički značajna razlika između skupina (p=0,52). Infekcije krvotoka su se u TKMS bolesnika javile značajno ranije (p=0,003), nego u OTJ bolesnika. Ovo je istraživanje potvrdilo nedavno objavljene podatke o povratku gram-negativnih patogena kao glavnih uzročnika infekcija krvotoka: gram-negativni bacili predominantno su izolirani u OTJ bolesnika, gdje su bili odgovorni za 58,5% slučajeva infekcija krvotoka, a učestalo su izolirani (39%) i kod TKMS bolesnika. Mikrobiološki izolati iz skupine bolesnika s transplantiranom jetrom pokazali su višu incidenciju rezistentnih enterobakterija koje proizvode beta-laktamaze proširenog spektra (ESBL) u usporedbi s izolatima bolesnika s transplantacijom krvotvornih matičnih stanica. P. aeruginosa bio je najkompliciraniji mikroorganizam za liječenje u obje skupine s obzirom da je rezistencija na karbapeneme ovog uzročnika iznosila 57% kod OTJ i 44% kod TKMS. U zaključku, infekcije krvotoka su važne komplikacije nakon ortotopne transplantacije jetre, kao i transplantacije krvotvornih matičnih stanica. Nadzor i analiza mikroorganizama koji uzrokuju infekcije krvotoka i druge ozbiljne infektivne komplikacije u transplantiranih bolesnika ostaje glavni preduvjet za planiranje intervencija vezanih uz liječenje infekcija u tih bolesnika.The aim of this retrospective study was to evaluate and compare the incidence, timing and etiology of bloodstream infections (BSIs) in patients treated with liver- (LT) or hematopoietic stem cell transplantation (HSCT) in a single institution. We evaluated 280 consecutive transplantations over a period of 34 months. Our results demonstrated 84 episodes of BSIs (47 in LT patients and 37 in HSCT patients) at a median of 28 days post-transplantation. Relative incidence of 34.6 and 29.4 BSI episodes per 100 LT and HSCT patients, respectively, did not differ significantly between the two groups (p=0.52). BSIs in HSCT patients occurred significantly earlier (p=0.003) than in LT patients. The recently described reemergence of gram-negative (GN) pathogens as causative agents of BSIs in these patients was confirmed: GN bacilli were the predominant isolates in the LT group, responsible for 58.5% of BSIs and a very frequent (39%) cause of BSIs in the HSCT group. A higher incidence of resistant enterobacteriaceae producing extended spectrum beta-lactamases was found in isolates from LT patients compared to HSCT patients. In both groups, Pseudomonas aeruginosa was the most difficult to treat organism, with 57% of these isolates in LT patients and 44% in HSCT patients being resistant to carbapenems. To conclude, BSIs were confirmed to be important infectious complications of both LT and HSCT. Surveillance and analysis of bacteria causing bloodstream and other serious infections in transplanted patients remain the main prerequisites for planning interventions regarding prevention and treatment of infections in these patient

Which questionnaires should we use to evaluate quality of life in patients with chronic graft-vs-host disease?

Aim To investigate the ability of two standard quality of life (QOL) questionnaires – The Short Form (36-item) Health Survey (SF-36) and The European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire- Core 30 (EORTC QLQ C30) to evaluate QOL in patients with chronic graft-vs-host disease (cGVHD) graded according to National Institutes of Health (NIH) consensus criteria. Methods In this cross-sectional study, QOL was assessed in patients who underwent allogeneic stem cell transplantation (allo-SCT) at the University Hospital Centre Zagreb and were alive and in complete remission for more than one year after allo-SCT. Results The study included 58 patients, 38 patients with cGVHD and 20 controls, patients without cGVHD. Patients with cGVHD scored according to the NIH criteria had significantly lower scores of global health status and lower QOL on all SF-36 subscales and most of QLQ C30 functional subscales (P < 0.050 for all comparisons). Furthermore, patients with active cGVHD had significantly lower QOL scores than patients with inactive cGVHD, and this difference was most evident in physical functioning subscale of SF-36 (P = 0.0007) and social functioning subscale of QLQ C30 (P = 0.009). Conclusion cGVHD scored according to the NIH criteria is correlated with patient-reported QOL, particularly in the physical domains as detected by SF-36. QLQ C30 questionnaire adds more information on social functioning and should be used as a valuable tool in the evaluation of social domains in cGVHD patients