8 research outputs found
Differences in staining for neutrophil elastase and its controlling inhibitor SLPI reveal heterogeneity among neutrophils in psoriasis
Differential expansion of circulating human MDSC subsets in patients with cancer, infection and inflammation
Background Myeloid-derived suppressor cells (MDSC) are a functional myeloid cell subset that includes myeloid cells with immune suppressive properties. The presence of MDSC has been reported in the peripheral blood of patients with several malignant and non-malignant diseases. So far, direct comparison of MDSC across different diseases and Centers is hindered by technical pitfalls and a lack of standardized methodology. To overcome this issue, we formed a network through the COST Action Mye-EUNITER (www.mye-euniter.eu) with the goal to standardize and facilitate the comparative analysis of human circulating MDSC in cancer, inflammation and infection. In this manuscript, we present the results of the multicenter study Mye-EUNITER MDSC Monitoring Initiative, that involved 13 laboratories and compared circulating MDSC subsets across multiple diseases, using a common protocol for the isolation, identification and characterization of these cells.
Methods We developed, tested, executed and optimized a standard operating procedure for the isolation and immunophenotyping of MDSC using blood from healthy donors. We applied this procedure to the blood of almost 400 patients and controls with different solid tumors and non-malignant diseases. The latter included viral infections such as HIV and hepatitis B virus, but also psoriasis and cardiovascular disorders.
Results We observed that the frequency of MDSC in healthy donors varied substantially between centers and was influenced by technical aspects such as the anticoagulant and separation method used. Expansion of polymorphonuclear (PMN)-MDSC exceeded the expansion of monocytic MDSC (M-MDSC) in five out of six solid tumors. PMN-MDSC expansion was more pronounced in cancer compared with infection and inflammation. Programmed death-ligand 1 was primarily expressed in M-MDSC and e-MDSC and was not upregulated as a consequence of disease. LOX-1 expression was confined to PMN-MDSC.
Conclusions This study provides improved technical protocols and workflows for the multi-center analysis of circulating human MDSC subsets. Application of these workflows revealed a predominant expansion of PMN-MDSC in solid tumors that exceeds expansion in chronic infection and inflammation
Effects of catechin and cyclophosphamide derivatives on leukemic cells
Galusan (-)-epigalokatechiny (EGCG) jest pochodn膮 katechiny najliczniej wyst臋puj膮c膮 w zielonej herbacie. Mafosfamid (MAF) jest pochodn膮 cyklofosfamidu i nale偶y do grupy zwi膮zk贸w alkiluj膮cych nowej generacji. Celem pracy by艂o okre艣lenie dzia艂ania EGCG i MAF na patologiczne kom贸rki hematopoetyczne. Badania wykonano na kom贸rkach ludzkiej promielocytarnej linii kom贸rkowej HL-60. Kom贸rki HL-60 eksponowano na dzia艂anie EGCG w dawkach 5 渭g, 10 渭g, 25 渭g, 50 渭g, 75 渭g, oraz 100 渭g w przeliczeniu na ml medium hodowlanego, natomiast MAF w dawkach 1 渭g, 2,5 渭g, 5 渭g, 10 渭g, 15 渭g, 20 渭g, 25 渭g i 50 渭g w przeliczeniu na ml medium hodowlanego. Analizowano tak偶e efekty kombinowanego dzia艂ania EGCG i MAF na kom贸rki leukemiczne. Funkcjonalne i morfologiczne zmiany zachodz膮ce w kom贸rkach HL-60 okre艣lono przy u偶yciu metody spektrofotometrii i cytometrii przep艂ywowej po 24 i 48 godzinach po ich potraktowaniu. Aktywno艣膰 metaboliczn膮 kom贸rek okre艣lono stosuj膮c test MTT (bromku 3[4,5-dimetylo-2-ilo]-2,5-difenylotetrazolu) i test FDA/PI (dioctan fluoresceiny / jodek propidyny), a wielko艣膰 kom贸rek i g臋sto艣膰 analizuj膮c warto艣ci FSC (ang. forward scatter channel) i SSC (ang. side scatter channel). Pochodne katechiny i cyklofosfamidu spowodowa艂y czasowe zmiany w aktywno艣ci metabolicznej kom贸rek okre艣lonej ilo艣ci膮 utworzonego formazanu przy zastosowaniu testu MTT oraz procentowymi ilo艣ciami kom贸rek w populacjach FDA+/PI-, FDA-/PI-, FDA-/PI+ przy u偶yciu testu FDA/PI, jak r贸wnie偶 czasowe zmiany w wielko艣ci i ziarnisto艣ci kom贸rek okre艣lane procentow膮 ilo艣ci膮 kom贸rek w populacjach FSC high/SSC low, FSC low/ SSC high, FSC high/SSC high. Przeciwleukemiczny potencja艂 EGCG i MAF by艂 zale偶ny od zastosowanego zwi膮zku, jego dawki oraz od punktu czasowego, w kt贸rym przeprowadzano analizy. W przypadku kombinowanego dzia艂ania EGCG i MAF na kom贸rki HL-60 zaobserwowano nasilenie aktywno艣ci przeciwnowotworowej. Wyniki przeprowadzonych bada艅 s膮 pierwszymi danymi ukazuj膮cymi efekty kombinowanego dzia艂ania mafosfamidu i galusanu (-)-epigalokatechiny na ludzkie kom贸rki leukemiczne.(-)-Epigallocatechin gallate (EGCG) is the major catechin found in green tea. Mafosfamide (MAF) is a cyclophosphamide derivative and belongs to the new generation alkylating agents. The aim of the study was to determine the alone and combined effects of EGCG and MAF on pathological hematopoitic cells.The investigations were conducted on human promyeolocytic HL-60 cells. The leukemic cells were exposed to the action of EGCG at doses 5 渭g, 10 渭g, 25 渭g, 50 渭g, 75 渭g, 100 渭g per 1 ml of medium and MAF at doses 1 渭g, 2,5 渭g, 5 渭g, 10 渭g, 15 渭g, 20 渭g, 25 渭g, 50 渭g per 1 ml of medium. The effects of EGCG and MAF on HL-60 cells were analyzed at 24 and 48 h after their exposure. The research was conducted using spectrophotometric and flow cytometric methods. The MTT assay, FDA/PI method and FSC?SSC measurement were used. After exposure of HL-60 cells to the EGCG and MAF the functional and morphological changes were observed. The various patterns of temporary alteration in the cell viability, cell membrane damage, the cell size and granularity were found. These are the first data that show the combined action of EGCG and MAF on human leukemic cells
The Legal Aspects of In Vitro Fertilisation (selected issues)
Przedmiotem niniejszej pracy s膮 prawne aspekty zap艂odnienia pozaustrojowego. Prac臋 rozpoczyna om贸wienie procedury in vitro oraz jej charakteru prawnego, czyli czy jest to lecznicza czynno艣膰 medyczna, czy nieterapeutyczna czynno艣膰 lekarska. Nast臋pnie zarysowane zosta艂y problemy prawne, kt贸re niesie ze sob膮 stosowanie tej metody. Stanowi艂o to przyczynek do poszukiwania rozwi膮za艅 prawnych w mi臋dzynarodowych regulacjach i orzecznictwie, ze szczeg贸lnym uwzgl臋dnieniem system贸w Rady Europy i Unii Europejskiej. Zaprezentowano tak偶e rozstrzygni臋cia przyj臋te w wybranych obcych porz膮dkach prawnych. W dalszej cz臋艣ci pracy dokonano analizy pocz膮tku ochrony 偶ycia i zdrowia ludzkiego na gruncie Konstytucji i prawa karnego w odniesieniu do embrionu in vitro. Opisane zosta艂y pr贸by uregulowania medycznie wspomaganej prokreacji, kt贸re by艂y podejmowane na przestrzeni ostatnich lat. Na ko艅cu om贸wione zosta艂y rozwi膮zania przyj臋te w ustawie o leczeniu niep艂odno艣ci.The subject of this thesis are the legal aspects of in vitro fertilisation. It disscusses the in vitro procedure and its legal nature, that is whether it is a therapeutical intervention or a non-therapeutical medical act. The legal problems of this method are then described. It was to look for some legal solutions in international law and judicature, especially in the Council of Europe and the European Union legal systems. The regulations in selected foreign countries are also presented. The rest of the dissertation focuses on the beginning of the protection of human life and health on the grounds of the Constitution and criminal law, with special emphasis on its implications for the fertilised embryo. The projects of legal regulation of the medically assisted procreation that have been practised in the past few years are also described. The thesis ends with an analysis of the solutions presented in the Infertility Treatment Act
Comparative safety of high-efficacy disease-modifying therapies in relapsingremitting multiple sclerosis : a systematic review and network meta-analysis
Secretory leukocyte protease inhibitor is present in circulating and tissue-recruited human eosinophils and regulates their migratory function
Eosinophils and secretory leukocyte protease inhibitor (SLPI) are both associated with Th2 immune responses and allergic diseases, but whether the fact that they are both implicated in these conditions is pathophysiologically related remains unknown. Here we demonstrate that human eosinophils derived from normal individuals are one of the major sources of SLPI among circulating leukocytes. SLPI was found to be stored in the crystalline core of eosinophil granules, and its dislocation/rearrangement in the crystalline core likely resulted in changes in immunostaining for SLPI in these cells. High levels of SLPI were also detected in blood eosinophils from patients with allergy-associated diseases marked by eosinophilia. These include individuals with eosinophilic granulomatosis with polyangiitis (EGPA) and atopic dermatitis (AD), who were also found to have elevated SLPI levels in their plasma. In addition to the circulating eosinophils, diseased skin of AD patients also contained SLPI-positive eosinophils. Exogenous, recombinant SLPI increased numbers of migratory eosinophils and supported their chemotactic response to CCL11, one of the key chemokines that regulate eosinophil migratory cues. Together, these findings suggest a role for SLPI in controlling Th2 pathophysiologic processes via its impact on and/or from eosinophils
Eosinophils regulate interferon alpha production in plasmacytoid dendritic cells stimulated with components of neutrophil extracellular traps
Differential expansion of circulating human MDSC subsets in patients with cancer, infection and inflammation
Background Myeloid-derived suppressor cells (MDSC) are a functional myeloid cell subset that includes myeloid cells with immune suppressive properties. The presence of MDSC has been reported in the peripheral blood of patients with several malignant and non-malignant diseases. So far, direct comparison of MDSC across different diseases and Centers is hindered by technical pitfalls and a lack of standardized methodology. To overcome this issue, we formed a network through the COST Action Mye-EUNITER (www.mye-euniter.eu) with the goal to standardize and facilitate the comparative analysis of human circulating MDSC in cancer, inflammation and infection. In this manuscript, we present the results of the multicenter study Mye-EUNITER MDSC Monitoring Initiative, that involved 13 laboratories and compared circulating MDSC subsets across multiple diseases, using a common protocol for the isolation, identification and characterization of these cells.
Methods We developed, tested, executed and optimized a standard operating procedure for the isolation and immunophenotyping of MDSC using blood from healthy donors. We applied this procedure to the blood of almost 400 patients and controls with different solid tumors and non-malignant diseases. The latter included viral infections such as HIV and hepatitis B virus, but also psoriasis and cardiovascular disorders.
Results We observed that the frequency of MDSC in healthy donors varied substantially between centers and was influenced by technical aspects such as the anticoagulant and separation method used. Expansion of polymorphonuclear (PMN)-MDSC exceeded the expansion of monocytic MDSC (M-MDSC) in five out of six solid tumors. PMN-MDSC expansion was more pronounced in cancer compared with infection and inflammation. Programmed death-ligand 1 was primarily expressed in M-MDSC and e-MDSC and was not upregulated as a consequence of disease. LOX-1 expression was confined to PMN-MDSC.
Conclusions This study provides improved technical protocols and workflows for the multi-center analysis of circulating human MDSC subsets. Application of these workflows revealed a predominant expansion of PMN-MDSC in solid tumors that exceeds expansion in chronic infection and inflammation