Remeikienė, Diana

Human platelet antigens (HPAs) are polymorphic structures on platelet membrane glycoproteins that may induce alloimmune responses after transfusion or pregnancy. Knowledge of HPA distribution in different populations is essential for improving transfusion practice and managing platelet transfusion refractoriness.

Background: The optimal number of CD34+ cells needed for repetitive autologous hematopoietic stem cell transplantation (auto-HSCT) in multiple myeloma (MM) patients remains unclear. As effective novel therapies for MM emerge, the use of salvage autologous hematopoietic stem cell transplantation (auto-HSCT) for relapse/refractory MM is questionable. Methods: We conducted a retrospective single center study using the clinical and laboratory databases. The study included 158 MM patients, 384 apheresis, 536 products and 211 auto-HSCT in the hospital of Lithuanian University of Health Sciences Kaunas Clinics from 2015 to 2022. Peripheral blood hematopoietic stem cells (PBHSC) were harvested after mobilization Cyclo+G-CSF (10 µg/kg) or an additional mobilizing agent such as plerixafor (0.24 mg/kg) if needed. PBHSC harvesting was done via apheresis. In our center the goal is to collect enough CD34+ cells for 3 auto-HSCT, which would equate to at least 10 x106 CD34+/kg. After each apheresis session, CD34+ counts were calculated. If the goal was achieved or if further mobilization was not possible, the harvesting was stopped. We calculated collected cells and divided patients in to 2 groups: A – if the goal is reached, B – goal is not reached. Descriptive statistics were performed on the variables of interest. We report frequencies and percentages as well as mean and ranges where applicable. Results: In total 158 MM patients were mobilized via 384 apheresis, that harvested 536 products, and transplanted at least 1 time. For 100 (63.3%) of the patients the CD34+ goal was achieved (A group), for 58 (36.7%) was not (B group). Mean collected CD34+ cell count for both groups were 12.45 x106/kg (range 2.0 x106 – 31.4 x106 /kg). To reach the CD34+ cell goal, 3 (3%) patients needed only 1 apheresis, most of them 67 (67%) needed 2 procedures, 28 (28%) – 3 apheresis procedures, and 2 (2%) patients needed 4 procedures. In comparison, in the B group more than half of the patients 35 (60.3%) needed 3 or more apheresis procedures. There is a correlation between CD34+ yield and number of apheresis procedures: patients yielding higher CD34+ counts required fewer apheresis procedures compared to those yielding lower CD34+ counts (p<0.01). Out of the patients that reached the targeted CD34+ goal (A group) transplanted in tandem 37 (37%) patients and 5 (5%) received as salvage auto-HSCT. Out of the B group 6 (10.2%) patients received tandem auto-HSCT, but no one received auto-HSCT as salvage. Conclusions: Our analysis evaluated the use of stored cryopreserved PBHSC for a salvage auto-HSCT in patients with MM. We showed that we could collect enough PBHSC for third auto-HSCT in most patients and usually they required 2 or 3 apheresis procedures. A third auto-HSCT was performed in just 5% of the included cohort. The increased survival rates offered by new cellular and immunotherapies, alongside the decreasing frequency of conducting salvage auto-HSCT warrant reconsideration of PBHSC collection goals.

Background and Objectives At the onset of the 21st century, tandem autologous hematopoietic stem cell transplantation (auto-HSCT) demonstrated efficacy in enhancing progression-free survival (PFS) and overall survival (OS) in patients with multiple myeloma (MM). Over recent decades, new effective drugs such as immunomodulators, proteasome inhibitors, and monoclonal antibodies have markedly improved treatment outcomes for MM, leading to increased PFS and OS. With the advent of new-generation drugs, the benefit of tandem auto-HSCT in the treatment of patients with newly diagnosed MM has become debatable. This study aimed to assess changes in stringent complete response (sCR) rates following the first and second auto-HSCT, as well as to evaluate the PFS and OS in relation to achieved sCR after tandem auto-HSCT. [...].

Background and Objectives The ideal quantity of CD34+ cells required for successive autologous hematopoietic stem cell transplantation (auto-HSCT) in multiple myeloma (MM) patients remains uncertain. With the advent of potent novel therapies for MM, the viability of employing salvage autologous hematopoietic stem cell transplantation (auto-HSCT) for relapsed or refractory MM is dubious. Material and Method Retrospective study was conducted at a single center, utilizing clinical and laboratory databases. The study encompassed 158 patients with multiple myeloma (MM), 384 apheresis procedures, 536 products, and 213 autologous hematopoietic stem cell transplants (auto-HSCT) at the Lithuanian University of Health Sciences Kaunas Clinics. Peripheral blood hematopoietic stem cells (PBHSC) were obtained through apheresis following mobilization with Cyclo+G-CSF (10 µg/kg), or an additional mobilizing agent like plerixafor (0.24 mg/kg) if necessary. Aim was to collect enough CD34+ cells for three auto-HSCT procedures, which amounted to at least 10 x106 CD34+/kg. CD34+ counts were assessed after each apheresis session, and harvesting ceased once the target was met or further mobilization was unfeasible. Descriptive statistics, including frequencies, percentages, means, and ranges where applicable, were used to analyze the variables of interest. Results 158 MM patients were mobilized via 384 apheresis, which yielded 536 products, and transplanted 213 times. To reach the CD34+ cell goal, 3 (1.9%) patients needed only 1 apheresis, most of them 67 (42.4%) needed 2 procedures, 28 (17.7%) – 3 apheresis procedures, and 2 (1.3%) patients needed 4 procedures, the rest of the patients 58 (36.7%) did not reach the goal. Mean collected CD34+ cell count in both groups were 12.45 x106/kg (range 2.0 x106 – 31.4 x106 /kg). Out of the patients that achieved the goal, only 4 (4%) patients received 3 transplantations in total (salvage auto-HSCT after tandem), 40 (40%) were transplanted two times out of them 29 (29%) were tandem transplantations, rest 11 (11%) was salvage auto-HSCT. In the cohort where patients did not achieve the goal only 7 (12.1%) patients were transplanted two times: 3 (5.2%) in tandem, 4 (6.9%) received as salvage after relapse. Conclusions and Recommendations Our study examined the utilization of stored cryopreserved PBHSC for salvage autoHSCT in MM patients. We demonstrated that we could obtain a sufficient quantity of PBHSC for a third auto-HSCT in the majority of patients, typically necessitating 2 or 3 apheresis procedures. However, only 4 patients (2.5%) in our study cohort underwent a third auto-HSCT. The improved survival rates associated with emerging cellular and immunotherapies, coupled with the declining frequency of salvage auto-HSCT, suggest the need to reassess PBHSC collection objectives.

Background: Peripheral blood stem cells (PBSCs) are widely used for autologous blood stem cell transplantation (auto-SCT) for patients with various conditions. CD34+ cell viability evaluation after cryopreservation and thawing is an important parameter used for quality assurance of the cryopreserved cells. CD34+ cell post-thaw viability results and their correlation with clinical parameters were evaluated. Methods: The study included 209 patients, 461 aphereses, 640 products and 239 auto-SCT in the Hospital of Lithuanian University of Health Sciences Kaunas Clinics from 2015 to 2022 December. The patient’s plasma and DMSO (final concentration 10%) were used for PBSC cryopreservation. PBSC products were cryopreserved using a controlled freezer and then transferred to vapor phase nitrogen. Cell viability was evaluated after thawing PBSC in a water bath at 37 °C. Immediately after thawing viability was assessed using a BD FACS Canto flow cytometer and BD Stem Cell Enumeration Kit. A linear regression model and Pearson correlation were used for statistical analysis. The study included 209 patients, 461 aphereses, 640 products and 239 auto-SCT in the Hospital of Lithuanian University of Health Sciences Kaunas Clinics from 2015 to 2022 December. The patient’s plasma and DMSO (final concentration 10%) were used for PBSC cryopreservation. PBSC products were cryopreserved using a controlled freezer and then transferred to vapor phase nitrogen. Cell viability was evaluated after thawing PBSC in a water bath at 37 °C. Immediately after thawing viability was assessed using a BD FACS Canto flow cytometer and BD Stem Cell Enumeration Kit. A linear regression model and Pearson correlation were used for statistical analysis. Results: In total 209 patients (106 females and 103 males) with a median age of 59 (range 18–73) years were harvested after G-CSF (10g/kg) or Cyclo+G-CSF or additional mobilizing agents such as plerixafor (0.24 mg/kg) for auto-SCT. The median post-thaw CD34+ viability was 75% (range 1–96). Median leukocytes (>1.0 × 109 /l) engraftment on two consecutive days was 11 (range 8–13) days, and 15 days for platelets (>50 × 109 /l), ranging 9–23 days after SCT. Statistically significant negative correlations were observed between CD34+ cell post-thaw viability and CD45+ count in leukapheresis product (p = 0.001, R2 = 0,076); CD34+ count (p = 0.002, R2 = 0.02); a volume of leukapheresis product (p = 0.001, R2 = 0.076). Conclusions: The results suggest that cryopreservation of cells using 10% DMSO final concentration is acceptable with successful cell engraftment in all patients. Lower CD45+ and CD34+ cell counts and lower leukapheresis volume had a better impact on post-thaw cell viability. Further studies are needed to confirm these findings and improve post-thaw cell viability.

The morbidity and mortality of BCR-ABL-negative myeloproliferative neoplasia (MPN) patients is highly dependent on thrombosis that may be affected by antiphospholipid antibodies (aPLA) and lupus anticoagulant. Our aim was to evaluate the association of the aPLA together with platelet receptor glycoprotein (GP) Ia/IIa c.807C>T CT/TT genotypes and thrombotic complications in patients with MPNs. The study included 108 patients with BCR-ABL-negative MPN with data of previous thrombosis. Two different screening and one confirmatory test for the lupus anticoagulant were performed. Thrombotic complications were present in 59 (54.6%) subjects. aPLA were more frequently found in MPN patients with thrombosis vs no thrombosis (25.4 and 6.1%; p = 0.007). MPN patients with arterial thrombosis were more frequently positive for aPLA vs no arterial thrombosis (38.8 and 11.9%; p = 0.001). aPLA were more frequently found in patients with cerebrovascular events vs other arterial thrombotic complications or no thrombosis, respectively (39.3, 6.1, and 12.9%; p < 0.001). MPN patients with thrombosis were more frequently positive with aPLA and had platelet receptor GP Ia/IIa c.807C>T CT/TT genotypes compared to MPN patients without thrombosis (18.6 and 2.0%; p = 0.006). aPLA alone or with coexistence with platelet receptor GP Ia/IIa c.807C>T CT/TT polymorphism could be associated with thrombotic complications in patients with MPN.

Background Human anti-SARS-CoV-2 convalescent plasma is used for the treatment of patients with coronavirus disease 2019 (COVID-19) who are at risk of severe illness. There is currently a lack of evidence about effectiveness of the human anti-SARS-CoV-2 convalescent plasma in the COVID-19 treatment. Method The retrospective analysis was consisted of data from 40 patients (19 men and 21 women; mean age 64.97±15.78) who were treated from COVID-19 in the hospital of the Lithuanian University of Health Sciences, Kaunas, Lithuania. The diagnosis of severe COVID-19 was confirmed by clinical symptoms and positive PCR test of nasopharyngeal swab. The participants treated with standard COVID-19 treatment (antiviral, immunomodulatory and antithrombotic therapy) combined with anti-SARS-CoV-2 convalescent plasma (250–500ml) with anti-spike (S) antibody titers of at least 1:600. Clinical outcomes were compared by 8-point disease severity scale (ranging from 0 [not infected] to 8 [death]), value of saturation (SpO2), the flow of oxygen therapy (O2), count of leucocytes (LEU) and lymphocytes (LY), c-reactive protein (CRP), prognostic laboratory markers (lactate dehydrogenase (LDH), ferritin, D-dimer) before and after 7-10 days of convalescent plasma transfusion. Results The flow of oxygen therapy decreased and the value of SpO2 improved significantly after 7-10 days (p<0.05). The count of LY, CRP and prognostic markers differed significantly (p<0.05). The score of disease severity scale did not differ before and after 7-10 days of convalescent plasma transfusion (p<0.05). Conclusion Treatment of severe coronavirus disease 2019 by adding convalescent plasma for treatment may remarkably improve clinical outcomes of these subjects.

Viena svarbiausių trombocitų funkcijų yra hemostazės užtikrinimas. Esant mažam trombocitų kiekiui kraujavimas gali prasidėti dėl minimalios traumos. [...].

Įvadas. Pasaulyje intensyviai plintanti SARS-CoV-2 infekcija, sukelianti ligą – COVID-19, ypač sunkias jos formas, neretai lemiančias letalias baigtis tampa našta visuomenei. Kartu vyksta efektyvių COVID-19 gydymo būdų paieška. Siekiama pasitelkti imuninio atsako savybes – panaudoti persirgusiųjų COVID-19 kraujyje susidariusius antikūnus, kurie imuninės plazmos transfuzijos būdu skiriami sergantiesiems COVID-19, kuriems yra sunkių ligos formų rizika bei reikalingas gydymas stacionare. Tyrimo tikslas. Įvertinti sergančiųjų sunkia COVID-19 gydymo imuninę pasveikusiųjų nuo COVID-19 plazma įtaką klinikinei ligos eigai. Tyrimo metodika. Retrospektyviai analizuoti sirgusių sunkia COVID-19 forma pacientų duomenys (19 vyrų ir 21 moteris, kurių vidutinis amžius – 64,97±15,78 metų), kurie buvo gydyti 2020 m. gruodžio mėn. – 2021 m. kovo mėn. . [...].

Background: Peripheral blood stem cells (PBSCs) are widely used for autologous blood stem cell transplantation (auto-SCT) around the world. Post-thaw viable CD34+ cell count is the important parameter for quality assurance of the cryopreserved cells. The CD34+ cell post-thaw viability was analysed and its relationship with clinical parameters was evaluated in this study. Methods: The study included 236 apheresis, 329 products and 106 hematopoietic SCT on 100 patients undergoing auto-SCT in the Hospital of Lithuanian University of Health Sciences Kaunas Clinics from 2015 to 2019 August. Patient’s plasma and DMSO (final concentration of 10%) was used for PBSC cryopreservation. All PBSC products were initially cryopreserved in a controlled freezer, then stored in vapor phase nitrogen. PBSC were thawed before use in water bath at 37°C. Viability was assessed immediately after thawing using BD FACS Canto flow cytometer and BD Stem Cell Enumeration Kit. A multivariate regression model and Pearson correlation were applied for statistical analysis. Results: In total 100 patients (51 females, 49 males) with a median age of 59 (range 18-73) years were harvested after G-CSF (10 g/kg) or Cyclo+G-CSF or additional mobilizing agents such as plerixafor (0,24mg/kg, n=4) for auto-SCT. Indications were multiple myeloma (n=82), non-Hodgkin lymphoma (n=15), germ-cell testicular tumor (n=1), sarcoma (n=1) and autoimmune encephalitis (n=1). Most patients received one auto-SCT. 24 patients received double SCT and 1 patient triple SCT. PBSC transplantations were performed for patients with the following diagnosis: 94 myeloma, 7 lymphoma, 3 germ-cell testicular tumors, 1 CNS lymphoma and 1 autoimmune encephalitis. The median post-thaw CD34+ viability was 79% (range 42-96). Median leukocytes (>1.0 x 109/l) engraftment on two consecutive days was 11 (range 8-13) days, and 15 days for platelets (>50 x 109/l), ranging 9-23 days after [...].