Anti-CD38 Therapy with Daratumumab for Relapsed/Refractory CD20-Negative Diffuse Large B-Cell Lymphoma.

Diffuse large B-cell lymphoma (DLBCL) is the most common and one of the most aggressive subtypes of non-Hodgkin's lymphomas. Front-line therapy consists of chemotherapy in combination with anti-CD20 monoclonal antibody rituximab. Relapses after rituximab-based regimen have poor prognosis and call for new treatment options. Immunohistochemistry analysis of relapsed DLBCL often reveal CD20-negative lymphoma, which limits repeated use of rituximab in combination with salvage chemotherapy. CD38 is a surface antigen that binds to CD38, CD31/PECAM-1 and hyaluronic acid. CD38 is an important mediator of signal transmission from the microenvironment into the cell. Anti-CD38 monoclonal antibody daratumumab has been approved for the treatment of multiple myeloma. Expression of CD38 on the surface of DLBCL is highly variable (compared to strong expression on myeloma cells), but can be easily assessed by flow cytometry or immunohistochemistry. A patient-derived xenograft (PDX) model of CD20-negative, CD38-positive DLBCL derived from a patient with rituximab-refractory DLBCL was used for in vivo experiments. We demonstrated that daratumumab suppressed growth of subcutaneous PDX tumours significantly more effectively than rituximab. Analysis of tumours obtained from mice treated with daratumumab revealed down-regulation of surface CD38, suggesting endocytosis of CD38-daratumumab complexes. The results suggest a potential clinical use of daratumumab in combination with salvage chemotherapy in patients with relapses of CD20-negative DLBCL. In addition, daratumumab might potentially serve as a suitable antibody moiety for derivation of antibodydrug conjugates for the targeted delivery of toxic payloads to the lymphoma cells.

Liver transplantation using grafts from donors after circulatory death - the first Czech Republic experience.

INTRODUCTION: Liver transplantation is established as a lifesaving procedure for patients with acute and chronic liver failure, as well as certain selected malignancies. Due to a continuing organ shortage and ever-growing patient waiting lists, donation after cardiac death (DCD) is becoming more frequently utilized in order to close the gap between “supply and demand”. METHODS: A retrospective analysis of DCD and subsequent liver transplantations was performed. RESULTS: From May 2016 to September 2019, a total of 9 DCD liver transplantations were performed in our institution. All cases except one were primary liver transplantations. The recipients comprised 5 (56%) males and 4 (44%) females. The mean DCD donor age was 41±12 (22-57) years, with ventilation duration of 7±1 days and warm ischemia time 19±3 minutes. The average recipient age was 51±22 (4-73) years, with an average cold ischemia 3h:59m±27m and manipulation time of 23±5 minutes. Periprocedural mortality was 1 (11%). Hepatitis C recurrence was documented in 1 (11%) patient. The mean follow-up time was 19±13 (7-37) months. Until now, we have not observed any signs of ischemic cholangiopathy. CONCLUSION: DCD liver transplantation allows us to enlarge the pool of potential liver grafts, thus decreasing the time spent on the liver recipient waiting list. This paper documents the first series of DCD liver transplantations in the Czech Republic.

[Deceased organ donors, legal regulations governing diagnosis of brain death, overview of donors and liver transplants in the Czech Republic]. / Zemrelí dárci orgánu, právní normy dia-gnostiky smrti mozku. Prehled dárcu a transplantací jater v Ceské republice.

The key restriction of transplantation medicine globally, as well as in the Czech Republic, concerns the lack of organs. The number of deceased donors, and thus the availability of organ transplants, has been stagnating in our country. The paper describes current legal regulations governing the dia-gnosis of brain death and primary legal and medical criteria for the contraindication of the deceased for organ explantation, gives an overview of the number of liver transplants, age structure, and diagnosis resulting in brain death of the deceased liver donors in the Czech Republic.

[Kidney transplantation from donors dying of methanol intoxication]. / Transplantace ledvin od dárcu zemrelých na intoxikaci metanolem.

INTRODUCTION: Intoxicated patients represent only a minimum of the total number of cadaveric donors; however, their significance within the transplant program in the Czech Republic has recently been emphasized in connection with the so-called methanol affair. MATERIAL AND METHODS: A retrospective analysis of methanol-poisoned patients who were declared brain-dead and subsequently underwent organ removal for transplantation purposes in the University Hospital Ostrava was performed. In cooperation with other transplant centres (Institute for Clinical and Experimental Medicine, Transplant Centre of the University Hospital Hradec Kráilové, Centre for Cardiovascular and Transplantation Surgery in Brno) the graft function, postoperative morbidity and mortality was evaluated. RESULTS: During the study period (since September 2012), organs from three brain-dead donors as a result of methanol intoxication were retrieved in the Transplant Centre of the University Hospital Ostrava. Subsequently, six kidneys were transplanted (3 males, 3 females). Postoperative complications occurred in two patients (33.3%), one patient died (16.7%). In one case (16.7%), the graft failed. None of the graft recipients developed clinical or laboratory signs of methanol poisoning. CONCLUSION: Death due to methanol intoxication is not a contraindication to organ donation, graft function and patient survival being comparable to organs from non-intoxicated donors.

Extracorporeal cardiopulmonary resuscitation-based approach to refractory out-of-hospital cardiac arrest: A focus on organ donation, a secondary analysis of a Prague OHCA randomized study.

BACKGROUND: Refractory out-of-hospital cardiac arrest (OHCA) has a poor outcome. In patients, who cannot be rescued despite using advanced techniques like extracorporeal cardiopulmonary resuscitation (ECPR), organ donation may be considered. This study aims to evaluate, in refractory OHCA, how ECPR versus a standard-based approach allows organ donorship. METHODS: The Prague OHCA trial randomized adults with a witnessed refractory OHCA of presumed cardiac origin to either an ECPR-based or standard approach. Patients who died of brain death or those who died of primary circulatory reasons and were not candidates for cardiac transplantation or durable ventricle assist device were evaluated as potential organ donors by a transplant center. In this post-hoc analysis, the effect on organ donation rates and one-year organ survival in recipients was examined. RESULTS: Out of 256 enrolled patients, 75 (29%) died prehospitally or within 1 hour after admission and 107 (42%) during the hospital stay. From a total of 24 considered donors, 21 and 3 (p = 0.01) were recruited from the ECPR vs standard approach arm, respectively. Fifteen brain-dead and none cardiac-dead subjects were ultimately accepted, 13 from the ECPR and two from the standard strategy group. A total of 36 organs were harvested. The organs were successfully transplanted into 34 recipients. All transplanted organs were fully functional, and none of the recipients died due to graft failure within the one-year period post-transplant. CONCLUSION: The ECPR-based approach in the refractory OHCA trial is associated with increased organ donorship and an excellent outcome of transplanted organs. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT01511666. Registered January 19, 2012.

[An overview of the results of renal transplantation in the Czech Republic]. / Piehled výsledku transplantací ledvin v Ceské republice.

Renal (kidney) transplantation is now a routine and the most successful form of renal replacement therapy. There is a long tradition of renal transplantation in the Czech Republic, The first was performed as early as 1961 in Hradec Kralove, and the programme as such was launched in 1966 with the first successful transplantation at the Institute of Experimental Surgery (later Institute for Clinical and Experimental Medicine, Prague). At present, transplantations are being performed at 7 transplantation centres (IKEM Prague, Centre for Cardiovascular and Transplantation Surgery Brno, Faculty Hospitals Hradec Kralove, Plzen, Olomouc and Ostrava and Faculty Hospital Motol for children). From the programme launch until the end of 2010, 8,761 renal transplantations were performed, 364 in 2010 alone. One-year patient and cadaver renal allograft survival, transplanted in the CR between 2000 and 2009, is around 95% and 92%, respectively, and 5-year survival is 87% and 81%, respectively. As of 31st December 2009, a total of 3,771 patients lived with functional renal allograft in the Czech Republic and the proportion of patients with irreversible renal failure treated with transplantation has recently been around 40%.

[Dual kidney transplantation]. / Duální transplantace ledvin.

Despite the increasing demand for organs for transplantation, the number of cadaveric donors remains stable and waiting time for transplantation is gradually getting longer. In addition to the options of using kidneys from living donors and those of non heart-beating donors (NHBD), an alternative approach is transplantation of both kidneys from adult marginal donors who would otherwise be considered unsuitable for single-kidney donation. Dual kidney transplantation involves the use of both kidneys from a marginal donor for a single recipient without the recipient having to cope with the drawbacks of a limited number of functioning nephrons. Normally. these kidneys would be excluded from the transplantation program and remain unused. The submitted presentation provides information on donor and recipient selection criteria and describes the course of the first dual kidney transplantation in an adult recipient in the Czech Republic.

Lack of Impact of Hyperchloremia in Brain-Dead Organ Donors on the Onset of Kidney Allograft Function in the Recipients.

BACKGROUND: Hyperchloremia produces renal vasoconstriction and fall in glomerular filtration rate. In 90% of brain-dead organ donors, diabetes insipidus develops, characterized by inappropriate diuresis, hyperosmolality, and hyperchloremia. The aim of this study was to determine the relationship between the serum concentration of chlorides of the donor and the onset of the function of the kidney allograft in the recipient. METHODS: We retrospectively studied 213 donors and kidney allograft recipients. Serum creatinine concentrations and glomerular filtration rates on the 1st, 7th, and 30th days after transplantation of the recipients from hyperchloremic donors were compared with the recipients from normochloremic donors, as well as the incidences of acute tubular necrosis and delayed graft function. RESULTS: On the 1st day, serum creatinine concentrations of the recipients from hyperchloremic and normochloremic donors, respectively, were 448.2 ± 212.1 µmol/L and 502.2 ± 197.8 µmol/L (P = .1), on the 7th day, 168.6 ± 102.6 µmol/L and 196.9 ± 120.6 µmol/L (P = .13), and on the 30th day, 129.4 ± 43.3 µmol/L and 131.8 ± 43.6 µmol/L (P = .73). The differences were statistically significant. The groups also did not differ significantly in glomerular filtration rates and incidences of acute tubular necrosis and delayed graft function. CONCLUSIONS: In this study, no significant correlation between serum chloride concentrations of the organ donors and the onset of the function of kidney allografts in the recipients was found.

Sensitivity to perioperative ischemia/reperfusion injury in male and female donor myocardium.

Many functions of the cardiovascular apparatus are affected by gender. The aim of our study was find out whether markers of cell death present in the donor myocardium differ in male and female hearts. The study involved 81 patients undergoing heart transplantation from September 2010 to January 2013. Patients were divided into two groups: male allograft (n=49), and female allograft (n=32). Two types of myocardial cell death were analyzed. High-sensitive cardiac troponin T as a necrosis marker and protein bcl-2, caspase 3 and TUNEL as apoptosis markers were measured. We observed a significantly higher level of high-sensitive cardiac troponin T after correcting for predicted ventricular mass in female donors before transplantation as well as in the female allograft group after transplantation throughout the monitored period (P=0.011). There were no differences in apoptosis markers (bcl-2, caspase 3, TUNEL) between male and female hearts before transplantation. Both genders showed a significant increase of TUNEL-positive myocytes one week after transplantation without differences between the groups. Moreover, there were no differences in caspase 3 and bcl-2 expression between the two groups. Our results demonstrated the presence of necrotic and apoptotic cell death in human heart allografts. High-sensitive cardiac troponin T adjusted for predicted ventricular mass as a marker of myocardial necrosis was higher in female donors, and this gender difference was even more pronounced after transplantation.

Immune characterization of the Lewis rats inoculated with K2 sarcoma cell line and newly derived R5-28 malignant cells.

Sarcoma is a relatively rare malignant disease with high mortality, bad prognosis and response to conventional therapy. Two possible models of this disease were tested: the K2 rat sarcoma cell line, which was described previously, and the new rat R5-28 cell line derived from a spontaneously growing rat neoplasm with sarcoma morphology. While all rats inoculated with K2 cells developed tumours at 22th-25th day after inoculation (D = 22-25), only 60%-75% of R5-28-inoculated rats were affected by tumours. The frequency and progress of the disease depended on the number of inoculated cells. No metastases were detected in both cases. All affected animals showed large splenomegaly. A possible response of some immune system components to tumours was tested. No tumour-infiltrating lymphocytes were revealed in the tumour tissue. Anti-tumour antibodies were not found in tumour-bearing animal sera. Appropriate changes in peripheral blood lymphocyte subsets were explored. While the relative numbers of both NK cells and Tc were impaired, no changes were noted in numbers of CD4+CD8- T helper cells. Leukocytosis with highly increased numbers of CD11b+ myeloid cells displaying variable expression of CD4 was detected in terminal stages of the disease.

[The risk of malignancy after organ transplantation]. / Riziko vzniku malignity po orgáinové transplantaci.

Improvements in immunosuppressive therapy during the past decade brought about improvements of the long term tolerance of organ allografts. However, the long-term immunosuppressive therapy has an important limitation, because it can increase the risk of cardivascular diseases, infections and tumors. As compared with age-matched healthy population, organ-transplant recipients have an increased incidence of tumors.

Proportion of glomerulosclerosis in procurement wedge renal biopsy cannot alone discriminate for acceptance of marginal donors.

BACKGROUND: The shortage of available kidneys for renal transplantation could be addressed, to some extent, by expanding the criteria for acceptance of marginal donors. The study of these criteria is limited by the selection of grafts actually retrieved and transplanted, therefore reduced to a study of risk factors. We have evaluated the potential of procurement renal biopies as an instrument for acceptance or refusal of donor kidneys for transplantation. METHODS: This was a prospective study of a consecutive series of 200 donors. Biopsies were performed by wedge technique at the donor operation and were evaluated for proportion of glomerulosclerosis, vascular and tubular changes, and interstitial fibrosis. The study included 387 renal grafts with a representative biopsy, transplanted, and followed-up for survival and functional evaluation; 24 hr creatinine clearance at 1 and 3 weeks, and 3, 6, 12, 18, and 24 months. RESULTS: Factors associated with initial graft function included cold ischemia time, number of DR mismatches, tubular changes, although donor age showed the strongest correlation with short- and long-term level of graft function. DR mismatches and retransplantation appeared to be the only significant risk factors for graft loss. The proportion of glomerulosclerosis (mean 8%, range 0-48%) correlated with graft function in the simple regression analysis. However, when age was taken into account glomerulosclerosis did not correlate significantly with graft function. Furthermore, glomerulosclerosis as high as 25% or more had an acceptable 3-year graft survival rate of 74.7%. CONCLUSION: Procurement biopsy provides only limited information for the decision whether or not to accept a kidney donor.

Y chromosome and vimentin used to trace the fate of allogeneic keratinocytes delivered to the wound by the recombined human/pig skin.

RHPS, composed of confluent allogeneic keratinocytes cultured on cell-free pig dermis, stimulates wound healing when applied with the keratinocyte layer facing the wound. So far it has not been clarified whether the confluent keratinocytes implanted 'upside-down' can 'take' or only stimulate healing by producing growth factors. Confluent male keratinocytes were grafted onto donor sites of three female patients. Biopsies were taken on days 4, 6 and 9 after grafting. The fate of donor cells was followed in paraffin sections by FISH for the Y chromosome and by persisting expression of vimentin taken as a marker of cultured keratinocytes. Histological evaluation was complemented by detection of keratin 10 and involucrin. All three donor sites healed within one week. On day 4 the early neoepidermis was multilayered but disordered after transplantation. A large proportion of cells were apparently of donor origin as indicated by the presence of Y chromosomes, irregular morphology, expression of vimentin in the bottom and upper layers of the neoepidermis, and by irregular expression of involucrin and keratin 10 only in the central layer of the neoepidermis. From day 6 onwards, the new epidermis acquired an ordered stratification. Involucrin and keratin 10 renewed normal distribution in suprabasal layers. Concomitantly, vimentin expression was decreasing. The Y chromosome was still found on day 6 but not on day 9. We concluded that confluent allogeneic keratinocytes temporarily 'take' to the wound and contribute to rapid wound closure, being replaced by the patient's epidermal cells after about one week.

Necrobiotic process causing burn wound conversion may be prevented by allogeneic keratinocytes delivered by the recombined human/pig skin.

The spontaneous necrobiotic process frequently causes conversion of DDB (deep 2nd degree wounds) into full-thickness skin loss (3rd degree wounds). We found that this process may be positively influenced by the activity of living human allogeneic keratinocytes cultured on acellular pig dermis. This RHPS, if applied 'upside-down' with the epidermal layer facing the wound, provides an opportunity for keratinocytes to influence the healing. The aim of the present study was to find conditions, in terms of timing and wound-bed preparation, for optimum healing activity of RHPS. The wound beds were prepared either with tangential excision, surface dermabrasion or deep dermabrasion. Out of 17 wounds grafted with RHPS after tangential excision, 15 (88%) healed in 4-10 days; early excised wounds (up to day 5) healed within less than 10 days after the injury. Out of 8 wounds grafted after surface dermabrasion, only 2 (25%) healed. Out of 6 wounds grafted with RHPS after deep dermabrasion, 4 (67%) healed. The optimum healing effect of RHPS and prevention of conversion was achieved in early tangentially excised wounds.

Two dynamic morphotypes of sarcoma cells, asymmetric stellate and triangle with leading lamella, are related to malignancy.

A notion of the dynamic morphotype was developed as a conjunction between cell shape and migration. This enabled the investigation of the relationship between malignancy and patterns of dynamic morphology in neoplastic cells in vitro. Time-lapse cinemicroscopy was used to analyse the cell behaviour of three rat neoplastic cell lines (K2, T15, and A8), differing in metastatic potential, that were instrumental in revealing a coincidence between high migratory activity and appearance of the 3D structure of actin cables in high-malignant A8 cells (Pokorná et al., 1994). A set of criteria was established for visual classification of cell morphology. Matching the pattern of cell morphology with locomotory activity led to identification of four dynamic morphotypes. Cell speed was determined by tracking and the dynamic morphotypes assigned by the operator. All the three cell populations were studied for incidence of the dynamic morphotypes in culture media differing in pH: 6.6 simulating acid extracellular condition in tumours, physiological 7.4, and alkaline 8.2. The results showed that acid pH stimulated motile activity in the intermediate-malignant T15 and most malignant A8 cells. The T15 and A8 cells also manifested a prolonged continuation of fast locomotion in the early G1 phase and displayed a prevalence of two fast moving dynamic morphotypes: asymmetric stellate and triangle with leading lamella.

[Monocytoid B-lymphocyte lymphoma: a new type within the spectrum of non-Hodgkin's B-cell lymphoma]. / Monocytoidný B-lymfocytový lymfóm: nová jednotka v spektre non-Hodgkinových lymfómov B-pôvodu.

BACKGROUND: Clonal proliferation of monocytoid B-lymphocytes (MBLy)--monocytoid B-cell lymphoma (MBCL) represents a "new" type of lymphoma within the spectrum of B-cell malignancies. OBJECTIVES: The aim of the study was to evaluate the possibilities of a routine histological and immunohistochemical diagnosis of MBCL. METHODS: Three cases of MBCL diagnosed in peripheral lymph nodes (n = 2) and in mammary gland with infiltration of regional lymph node (n = 1) were analyzed both histologically and immunohistochemically using a panel approach (Ig chains, CD30 antigen, markers of B-cells, T-cells and of monocytes/histiocytes). RESULTS: Morphological appearance of neoplastic cells of MBCL is identical to that of MBLy in reactive conditions--kidney-shaped nuclei, bright clear PAS-negative cytoplasm, and small inconspicuous nucleoli. CONCLUSIONS: Morphological appearance together with immunophenotypic results (positivity of CD20 and Ki-B5, negativity of CD3, CD43, CD45RO, and of lysozyme, negativity of CD30) are considered to represent sufficient diagnostic criteria of MBCL, including its differential diagnosis of other B-cell low grade malignancies. An increase of large cell type MBLy might represent a feature of a secondary blastic transformation of MBCL. (Tab. 2, Fig. 5, Ref. 27.)

[Techniques, tactics and organizational aspects of distant procurement of donor hearts for transplantation. Experience at the Department of Cardiovascular and Transplantation Surgery of IKEM with 100 distant procurement of donor hearts]. / Technické, taktické a organizacní aspekty vzdáleného odberu dárcovského srdce k transplantaci. (Zkusenosti KKTCH IKEM se 100 vzdálenými odbery srdce)

Transplantation of the heart is currently an accepted therapeutic method. One of the factors which have an adverse effect on its availability is shortage of suitable donors. Distant procurement of hearts helps to extend the number of organs for transplantations. The authors present their experience with distant procurement of donor hearts performed during more than three years. They evaluate the influence of cold ischaemia time of the cardiac graft on early results of cardiac transplantations. In the discussion they try to draw attention to some problems associated with this technique for other organ retrieval teams participating in multiple organ procurement.

[Techniques and tactics in multiorgan harvesting]. / Technika a taktika multiorgánového odberu.

The eighties and nineties are characterized by potent development of transplantology. Despite this the number of patients with organ failures waiting for a suitable organ is steadily increasing. Due to the permanent shortage of donors transplantation surgery tries to implement the maximum possible multiple organ collection. In the conclusion the authors emphasize the importance of satisfactory cooperation of the anaesthesiologist attending donors and the organ collection teams.

Heart transplantations at the Heart Center IKEM, Prague, Czech Republic.

The heart transplant program at the Institute for Clinical and Experimental Medicine in Prague was established on January 31, 1984. Through November 2012, 881 orthotopic cardiac transplantations have been performed, with an annual rate of about 40 procedures. Current legislation concerning solid organ transplantations in the Czech Republic is described. Like other centers, we have noticed an increasing age of donors, and, reflecting the shortage of grafts, we have expanded our selection criteria for heart transplantation. The advent of a mechanical circulatory support program at our center in April 2003 has given us another valuable tool in the management of chronic heart failure patients. Currently, around half of our patients are transplanted from mechanical support. Nonischemic etiology of heart failure is a leading cause of transplantation at our center, followed by ischemic cardiomyopathy, valvular heart lesions, and adult congenital heart diseases. Our current immunosupression protocol, including induction therapy, is outlined in detail and survival rates, as well as most common complications and our treatment strategies, are also discussed.