Nádory z blastických plazmocytoidních dendritických buněk představují raritní hematologickou malignitu s agresivním průběhem a nepříznivou prognózou. Pro onemocnění jsou charakteristické primární kožní léze, často se současným postižením kostní dřeně a extramedulárních orgánů. Diagnostika je založena na imunohistochemickém průkazu a potvrzení ko-exprese znaků CD4 a CD56 s variabilním vyjádřením dalších markerů specifických pro dendritické buňky pomocí průtokové cytometrie. Medián věku v době diagnózy je 60–70 let, onemocnění se 3krát častěji vyskytuje u mužů. Poznatky o léčbě jsou omezeny na retrospektivní analýzy malých souborů nemocných. Navzdory obecně dobré iniciální léčebné odpovědi na chemoterapii dochází v odstupu řádu měsíců k časnému relapsu a následné rychlé systémové diseminaci. Publikovaná data ukazují na vyšší účinnost indukční léčby pro akutní lymfoblastickou leukemii následovanou alogenní transplantací krvetvorných buněk ve srovnání s jinými režimy indukční, respektive konsolidační chemoterapie. Předkládaný článek přináší kazuistiku 22letého nemocného, u něhož byla stanovena diagnóza nádoru z plazmocytoidních dendritických buněk s izolovaným kožním postižením. Pacient podstoupil indukční a jeden cyklus konsolidační chemoterapie podle protokolu GMALL pro akutní lymfoblastickou leukemii, následovanou alogenní transplantací krvetvorných buněk od nepříbuzenského dárce v 1. kompletní remisi onemocnění. Podání chemoterapie i provedení transplantace proběhlo bez závažných komplikací a nemocný zůstává v remisi onemocnění s dobrou kvalitou života v době vzniku této práce, 22 měsíců od alogenní transplantace. Kazuistika dokumentuje úspěšný léčebný přístup k prognosticky nepříznivému onemocnění. Článek je doplněný o literární přehled diagnostiky, klinického obrazu a léčebných strategií u pacientů s nádory z blastických plazmocytoidních dendritických buněk. Cílem sdělení je také zvýšit povědomí o této vzácné nozologické jednotce, a přispět tak k rychlejší diagnostice a včasnému zahájení intenzivní terapie nutné ke kontrole průběhu nemoci.
Blastic plasmacytoid dendritic cell neoplasm represents a rare hematologic malignancy of aggressive biology and unfavourable prognosis. This disease is characterised by primary skin lesions often followed by bone marrow and extramedullary involvement. Diagnosis is based on immunohistochemistry and flow cytometric demonstration of CD4 and CD56 co-expression with variable positivity of other markers specific to dendritic cells. The median age at diagnosis ranges from 60 to 70 years, with males affected 3 times more frequently. Knowledge of its treatment is limited to retrospective analyses of small patient cohorts. Despite generally favourable treatment response to chemotherapy, the disease tends to relapse in a few months with rapid systemic dissemination. The reported data suggest that induction therapy for acute lymphoblastic leukaemia followed by allogeneic hematopoietic cell transplantation is more effective than other induction or consolidation chemotherapy strategies. The article reports the case of a 22-year old male with newly diagnosed blastic plasmacytoid dendritic cell neoplasm with skin-limited symptoms. The patient underwent induction and one cycle of consolidation chemotherapy accordingto the GMALL protocol for acute lymphoblastic leukaemia followed by allogeneic hematopoietic cell transplantation in the first complete remission. Chemotherapy and transplantation were not associated with serious complications and the patient remains in remission with a good quality of life, 22 months after allogeneic transplantation. This case details the successful treatment strategy in a disease with unfavourable prognosis. The article also presents a review of the literature regarding diagnosis, clinical presentation and treatment strategies in patients with blastic plasmacytoid dendritic cell neoplasm. The aim of the paper is also to provide information about this rare nosological unit and promote prompt diagnosis and treatment initiation necessary for disease control.
- Klíčová slova
- nádor z blastických plazmocytoidních dendritických buněk,
- MeSH
- diagnostické techniky a postupy MeSH
- hematologické nádory * diagnóza terapie MeSH
- imunofenotypizace MeSH
- indukční chemoterapie metody MeSH
- lidé MeSH
- mladý dospělý MeSH
- transplantace kostní dřeně MeSH
- výsledek terapie MeSH
- vzácné nemoci diagnóza terapie MeSH
- Check Tag
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
1 svazek : ilustrace ; 30 cm
Here, we propose to prepare mRNA vaccine against a number of candidate immunogenic CMV proteins. The vaccine will include antigen presenting cells electroporated mRNA coding for viral protective antigens and immunostimulatory molecules known to enhance activation of T-cells directed to Th1. To evaluate the protective potential of each CMV vaccine component the magnitude of CMV T-cell response of seropositive donors or patients after HSCT to each individual protein will be monitored and analyzed in the context of CMV DNA levels of HSCT patient. Tcell response will be characterized phenotypicaly and functionally by multiparametric flow cytometry. The result of the study will enable to elaborate immunotherapy protocol under good manufacturing practise conditions. mRNA vaccine will be also used for monitoring in CMV-specific T cell response of patients after HSCT.
Navrhujeme konstrukci mRNA vakcíny proti CMV, která bude sloužit ke stimulaci a namnožení T lymfocytů specifických pro hlavní protektivní virové antigeny a bude využitelná pro monitorování stavu imunity proti CMV u pacientů po transplantaci hematopoetických krevních buněk případně k expanzi T-buněk specifických pro CMV ex vivo pro terapeutické účely. Vakcína bude obsahovat antigen prezentující buňky do nichž se elektroporací vnese rekombinantní mRNA, kódující několik virových antigenů a imunostimulační molekuly usměrňující buněčnou imunitní odpověď k typu Th1. Budeme analyzovat odpověď dárců kmenových buněk, pacientů po HSCT včetně průběhu jejich hladiny DNA CMV, což povede k výběru virových antigenů, které přispívají k protektivitě alespoň u 70% osob bez ohledu na jejich HLA haplotyp. Nalezneme podmínky stimulace T lymfocytů touto vakcínou in vitro, což bude základem pro další vývoj léčivého přípravku v podmínkách správné laboratorní praxe.
Among malignant diseases, chronic myeloid leukaemia (CML) is one of the best suited candidates for immunotherapy. For this purpose it is necessary to broaden the present knowledge on the immunology of this disease. As a part of such a project, the levels of kynurenine (KYN) and neopterin (NPT) were studied in 28 CML patients and in the same number of healthy subjects. At diagnosis, both KYN and NPT levels were found to be elevated in a significant portion of the patients and dependent on their leukocyte count. As in the case of KYN, increased NPT levels dropped after achieving remission. When correlating KYN and NPT levels with a selection of other markers tested, significant association was revealed only in the case of CRP and IL-6. However, there were several patients with increased KYN levels in whom NPT was not detected, and vice versa. The relapse of the disease observed in two patients was accompanied by an increased level of NPT in both cases, but by an increased level of KYN in only one of them. No significant correlation was found between KYN and NPT levels in sera taken at diagnosis. However, when the whole set of sera was taken into consideration, the association became statistically significant. Although the data obtained revealed a number of similarities between KYN and NPT production in CML patients, it also suggested a difference in the kinetics of these two biomarkers' production.
- MeSH
- biologické markery krev MeSH
- C-reaktivní protein metabolismus MeSH
- chronická myeloidní leukemie krev MeSH
- dospělí MeSH
- interleukin-6 krev MeSH
- kynurenin krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- lineární modely MeSH
- mladý dospělý MeSH
- neopterin krev MeSH
- počet leukocytů MeSH
- senioři MeSH
- studie případů a kontrol MeSH
- tryptofan krev MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
1 svazek : ilustrace, tabulky ; 30 cm
Determination of the effects of chronic myeloid leukemia (CML) on immune profile of patient in dependence of its treatment modalities and the results of its treatment. Examination of leukemic cells, sampled prior to any therapy, for the presence of 15-20 selected antigens, determination of the cellular and humoral immunity to these antigens, and correlation of the findings with the clinical picture. An attempt to determine the optimal time for administering the anti-tumor vaccine-to-be using a mathematical model recently constructed.
Určení vlivu chronické myeloidní leukémie (CML) na imunitní profil pacienta v závislosti na způsobu a výsledků léčby. Stanovení antigenního složení leukemických buněk odebraných před zahájením terapie a určení buněčné a humorální imunity proti těmto antigenům a korelace nálezů s klinickým obrazem. Vytvoření matematického modelu pro určení optimální doby pro podání budoucí protinádorové vakcíny. Studie by měla objasnit komplex problémů, které se týkají dosud neznámých aspektů interakce mezi nádorovými buňkami a imunitním systémem pacientů s CML. Tyto poznatky by měly dále pootevřít dveře pro vývoj vakcíny k terapii této nemoci a snad i dalších myeloidních malignit. Znalost imunitního stavu jednotlivých pacientů umožní personalizovat jiné terapeutické postupy a tak přispěje k optimalizaci léčby.
Aurora kinase A (AURKA) is a centrosomal protein that is overexpressed in a number of human malignancies and can contribute to tumor progression. As we used this protein as a target of DNA immunization, we increased its immunogenicity by the addition of the PADRE helper epitope and decreased its potential oncogenicity by mutagenesis of the kinase domain. For in vitro analysis of induced immune responses in mice, we identified the Aurka(220-228) nonapeptide representing an H-2Kb epitope. As DNA vaccination against the Aurka self-antigen by a gene gun did not show any antitumor effect, we combined DNA immunization with anti-CD25 treatment that depletes mainly regulatory T cells. Whereas 1 anti-CD25 dose injected before DNA vaccination did not enhance the activation of Aurka-specific splenocytes, 3 doses administered on days of immunizations augmented about 10-fold immunity against Aurka. However, an opposite effect was found for antitumor immunity-only 1 anti-CD25 dose combined with DNA vaccination reduced tumor growth. Moreover, the administration of 3 doses of anti-CD25 antibody alone accelerated tumor growth. Analysis of tumor-infiltrating cells showed that 3 anti-CD25 doses not only efficiently depleted regulatory T cells but also activated helper T cells and CD3(-)CD25(+) cells. Next, we found that blockade of the PD-1 receptor initiated 1 week after the first immunization was necessary for significant inhibition of tumor growth with therapeutic DNA vaccination against Aurka combined with depletion of CD25 cells. Our results suggest that combined cancer immunotherapy should be carefully evaluated to achieve the optimal antitumor effect.
- MeSH
- aktivace lymfocytů MeSH
- antigeny CD279 antagonisté a inhibitory MeSH
- aurora kinasa A genetika imunologie metabolismus MeSH
- buňky - růstové procesy účinky léků MeSH
- CD8-pozitivní T-lymfocyty imunologie MeSH
- DNA vakcíny MeSH
- epitopy T-lymfocytární genetika imunologie metabolismus MeSH
- H-2 antigeny metabolismus MeSH
- HEK293 buňky MeSH
- imunizace MeSH
- imunoterapie * MeSH
- kombinovaná terapie MeSH
- lidé MeSH
- monoklonální protilátky terapeutické užití MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- protinádorové vakcíny imunologie MeSH
- receptor interleukinu-2 - alfa-podjednotka imunologie MeSH
- regulační T-lymfocyty imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Indoleamine 2,3-dioxygenase 1 (IDO1), IDO2 and tryptophan 2,3-dioxygenase (TDO) represent some of the key immune regulators. Their increased activity has been demonstrated in a number of human malignancies but not yet in chronic myeloid leukemia (CML). In the present study, the activity of these enzymes was tested in 29 CML patients and 28 healthy subjects by monitoring the kynurenine (KYN)/tryptophan ratio. Serum samples taken prior to the therapy displayed a highly significant difference in KYN levels between the patient and control groups. However, increased KYN levels were detected in only 13 (44.8%) of these CML patients. The KYN levels in pretreatment sera of the patients correlated with the tumor burden. There was also a strong correlation between KYN levels and uric acid levels (UA). This suggests but does not prove the possible involvement of UA in activating IDO family of enzymes. Whenever tested, the increased KYN levels normalized in the course of the therapy. Patients with normal KYN levels in their pretreatment sera and subsequently treated with interferon-α, showed a transitory increase in their KYN levels. The present data indicate that CML should be added to the malignancies with an increased activity of the IDO family of enzymes and suggest that IDO inhibitors may be used in the treatment of CML patients.
- MeSH
- biologické markery krev MeSH
- chronická myeloidní leukemie * MeSH
- dospělí MeSH
- indolamin-2,3,-dioxygenasa MeSH
- kynurenin * krev MeSH
- kyselina močová * MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- tryptofan * krev MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
The asparaginyl endopeptidase legumain that is overexpressed in M2-polarized tumor-associated macrophages has been identified as a suitable target for elimination of these cells supporting tumor progression. To enhance the efficacy of DNA immunization against legumain, we performed several modifications in this protein that could improve induction of immune responses. First, we mutated the RGD motif into GGD or RGG sequences. This alteration resulted in diminished maturation of legumain and impaired cellular localization. Then, as tolerance to self-antigens can be broken by the activation of CD4 T-cell help, we tried to enhance the immunogenicity of legumain by the insertion of a foreign helper epitope, namely the p30 epitope from the tetanus toxin. Finally, the 2 modifications were combined. After gene gun DNA immunization of C57BL/6 mice with these constructs, we identified the Lgmn111-119 CD8 T-cell epitope that binds to H-2D molecules. Furthermore, we showed that mutagenesis in the RGD motif significantly enhanced the immune response against legumain. The addition of the p30 helper epitope induced the specific production of IFN-γ by T cells, but did not significantly increase legumain-specific immunity activated after mutagenesis in the RGD motif which might be caused by simultaneous activation of a Th2 response demonstrated by the production of IL-4. However, the beneficial effect of the helper epitope on legumain-specific response was proved after the depletion of regulatory T cells by antibody against CD25 that preferentially stimulated Th1 immunity. The antitumor effect of the modified legumain gene was shown in the immunization against tumors induced by MK16 cells.
- MeSH
- aminokyselinové motivy genetika MeSH
- biolistika MeSH
- buňky NIH 3T3 MeSH
- CD8-pozitivní T-lymfocyty imunologie MeSH
- cysteinové endopeptidasy genetika metabolismus MeSH
- DNA vakcíny * MeSH
- epitopy T-lymfocytární genetika metabolismus MeSH
- experimentální nádory imunologie terapie MeSH
- HEK293 buňky MeSH
- imunoterapie metody MeSH
- interferon gama metabolismus MeSH
- lidé MeSH
- makrofágy imunologie MeSH
- mutace genetika MeSH
- mutageneze cílená MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové biomarkery genetika metabolismus MeSH
- peptidové fragmenty genetika metabolismus MeSH
- protinádorové vakcíny * MeSH
- T-lymfocyty pomocné-indukující imunologie MeSH
- tetanový toxin genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Since its discovery, DNA vaccination has become an effective strategy for the development of vaccines against cancer including cervical carcinoma (CC). The formation of CC is associated with human papillomavirus (HPV) infection. Viral E6 and E7 oncoproteins are suitable targets for therapeutic vaccination. To adapt the HPV16 E6 oncogene for DNA immunisation, we performed several modifications. First we fused the E6 gene with the 5' or 3'-terminus of the Escherichia coli beta-glucuronidase (GUS) gene and showed enhanced immunogenicity of the 3' fusion (GUS.E6). Then, as the E6 oncogene contains two alternative introns that result in the production of truncated forms of the E6 protein, we abolished the 5' splice site in the E6 gene. This modification completely eliminated the expression of the truncated E6 transcripts and thus increased the production of the full-length E6 protein. At the same time, it moderately reduced the immunogenicity of the modified non-fused (E6cc) or fused (GUS.E6cc) genes, probably as a consequence of the substitution in the immunodominant E6 epitope following the abolishment of the splice site. Furthermore, we reduced the oncogenicity of the E6 protein by two point mutations (E6GT) that, together, prevented E6-mediated p53 degradation. Finally, we constructed the GUS.E6GT gene characterized by enhanced safety and immunogenicity when compared with the wild-type E6 gene.
- MeSH
- bodová mutace MeSH
- buněčné linie MeSH
- cytokiny biosyntéza MeSH
- DNA vakcíny genetika imunologie MeSH
- exprese genu MeSH
- glukuronidasa genetika imunologie MeSH
- introny MeSH
- leukocyty mononukleární imunologie MeSH
- lidé MeSH
- lidský papilomavirus 16 genetika imunologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádory prevence a kontrola MeSH
- onkogenní proteiny virové genetika imunologie MeSH
- proteiny z Escherichia coli genetika imunologie MeSH
- protilátky virové krev MeSH
- rekombinantní fúzní proteiny genetika imunologie MeSH
- represorové proteiny genetika imunologie MeSH
- vakcíny proti papilomavirům genetika imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A series of DNA vaccines based on the bcr-abl fusion gene were developed and tested in mice. Two mouse (BALB/c) bcr-abl-transformed cell lines, B210 and 12B1, which both expressed p210bcr-abl and were oncogenic for syngeneic animals but differed in some other respects, were used as a model system. In the first series of experiments, plasmids carrying either the complete bcr-abl fusion gene or a fragment thereof coding for a 25-amino acid-long junction zone (bcr-abl25aa) linked with genes coding for a variety of immunostimulatory factors were used as the DNA vaccines. A plasmid carrying the complete bcr-abl gene was capable of inducing protection against challenge with either B210 or 12B1 cells. However, the DNA vaccines based on the gene fragment coding for p25aabcr-abl did not induce significant protection. To localize the immunizing epitopes on the p210bcr-abl protein, the whole fusion gene was split into nine overlapping fragments and these, individually or in various combinations, were used for immunization. Although none of the vaccines based on any single fragment provided potent protection, some combinations of these fragment-based vaccines were capable of eliciting protection comparable to that seen after immunization with the whole-gene vaccine. Surprisingly, a mixture of six fragment-vaccines was more immunogenic than the complete set of fragment DNA vaccines. To analyze this phenomenon, the three fragments missing from the hexavaccine were either individually or in various combinations mixed with the hexavaccine. The results obtained suggested that the product of the fragment coding for 197 amino acids forming the N-terminal of the BCR protein was involved in the decreased immunogenicity. However, further experiments are needed to clarify the point. Additional experiments revealed that all the important epitopes were located in the ABL portion of the p210bcr-abl protein. The livers, spleens and bone marrows of the successfully immunized animals were tested for the presence of bcr-abl-positive cells by RT-PCR. The results were negative, this suggesting that these animals were free of any residual disease.
- MeSH
- bcr-abl fúzové proteiny genetika imunologie MeSH
- časové faktory MeSH
- chronická myeloidní leukemie genetika imunologie patologie prevence a kontrola MeSH
- DNA vakcíny genetika imunologie MeSH
- HL-60 buňky MeSH
- imunizace MeSH
- lidé MeSH
- mapování epitopu MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- peptidové fragmenty imunologie MeSH
- protinádorové vakcíny genetika imunologie MeSH
- transfekce MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Two mouse HPV16-transformed cell lines, viz. MK16 cells, which induce metastasizing tumors, and TC-1 cells, which induce non-metastasizing tumors were transduced with the gene for mouse endostatin. Two clones constitutively expressing endostatin were isolated from each of them. They were denoted ME3 and ME9, and TE2 and TE5, respectively. When inoculated into mice, ME3 cells were non-oncogenic. Nearly all mice inoculated with ME9 cells developed tumors, but considerably later than did the parental MK16 cells and metastasis formation was strongly reduced in these animals. On the other hand, TE2 and TE5 cells displayed oncogenic potential similar to that of the parental cells. To provide more information on these different effects of endostatin production, cell lysates of all six lines studied were tested for the content of 25 factors known to be involved in angiogenesis. The parental MK16 cells differed from the parental TC-1 cells and also from all endostatin producing sublines by a markedly higher production of interleukin 1alpha (IL-1alpha) and, to a lesser extent, by a higher production of several other factors tested. Additional experiments indicated that the suppression of the production of IL-1alpha by the parental MK16 caused by endostatin was due to an autocrine mechanism.
- MeSH
- autokrinní signalizace MeSH
- časové faktory MeSH
- endostatiny genetika metabolismus MeSH
- endoteliální buňky metabolismus MeSH
- geny ras MeSH
- interleukin-1alfa metabolismus MeSH
- interleukin-2 genetika metabolismus MeSH
- kultivační média speciální metabolismus MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorová transformace buněk MeSH
- nádory plic genetika imunologie metabolismus prevence a kontrola sekundární virologie MeSH
- onkogenní proteiny virové genetika MeSH
- Papillomavirus E7 - proteiny MeSH
- proliferace buněk MeSH
- represorové proteiny genetika MeSH
- transdukce genetická MeSH
- transformované buněčné linie MeSH
- virová transformace buněk MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH