Cíl: Zaměřili jsme se na stanovení prevalence infekce SARS-CoV-2 se symptomatickým nebo asymptomatickým průběhem a na identifikaci prediktorů symptomatické nebo asymptomatické infekce SARS-CoV-2 u pacientů během sedmi měsíců následujících po transplantaci alogenních hematopoetických kmenových buněk (alo-HSCT) v období cirkulace varianty omikron. Metody: Prevalence proběhlé infekce SARS-CoV-2 byla detekována u pacientů během sedmi měsíců po allo-HSCT v omikronovém období pomocí buněčné a humorální imunitní odpovědi proti nukleoproteinu SARS-CoV-2 (NCP). Výsledky: Pozitivní markery prodělané infekce byly identifikovány u 45,2 % pacientů (n = 42). Infekce byla asymptomatická u 68,4 % pacientů s anti-NCP pozitivitou. Hledání rizikových faktorů pro symptomatickou infekci SARS-CoV-2 u příjemců alo-HSCT odhalilo, že nízká úroveň rekonstituce B buněk byla jediným signifikantně souvisejícím rizikovým faktorem. Závěr: Vysoký podíl příjemců alo-HSCT, kteří byli asymptomaticky infikováni do sedmi měsíců po transplantaci v letech 2022–2023, přestože byli imunokompromitovaní a neočkovaní, ukazuje na oslabení cirkulujícího viru a může signalizovat pro pacienty po transplantaci menší riziko onemocnění SARS-CoV-2 v omikronovém období. Ukázalo se, že očkování těchto pacientů proti SARS- -CoV-2 je spojeno s nízkým, ale významným rizikem exacerbace vyléčené chronické reakce štěpu proti hostiteli (GVHD – Graft Versus Host Disease) a s rizikem de novo GVHD. Nízká úroveň rekonstituce B-buněk byla jediným významným rizikovým faktorem pro symptomatickou infekci SARS-CoV-2 u příjemců alo-HSCT.
Aim: We aimed to determine the prevalence of SARS-CoV-2 infection, including both symptomatic and asymptomatic courses, and to identify predictors of asymptomatic or symptomatic SARS-CoV-2 infection in patients within seven months after allo-HSCT (allogenic hematopoietic stem cell transplantation) in the Omicron period. Methods: Prevalence of the past SARS-CoV-2 infection was determined in patients within seven months after allo-HSCT in the Omicron period using the cellular and humoral immune response against the SARS-CoV-2 nucleoprotein (NCP). Results: Positive markers of past infection were identified in 45.2% of patients (n = 42). The infection was asymptomatic in 68.4% of anti-NCP positive patients. The search for risk factors for symptomatic SARS-CoV-2 infection in allo-HSCT recipients revealed that a low level of B cell reconstitution was the only significantly associated risk factor. Conclusion: A high proportion of allo-HSCT recipients who were asymptomatically infected within up to seven months after transplantation from 2022 to 2023 despite being immunosuppressed and unvaccinated indicates an attenuation of the circulating virus and may signal less risk for transplanted patients from SARS-CoV-2 infection in the Omicron period. Vaccination of these patients against SARS-CoV-2 was shown to be associated with a low but significant risk of exacerbation of cured chronic GVHD (graft versus host disease) and the risk of de novo GVHD. The low level of B-cell reconstitution was the only significant risk factor for symptomatic SARS-CoV-2 infection in HSCT recipients.
- Klíčová slova
- Omikron,
- MeSH
- asymptomatické infekce * epidemiologie MeSH
- B-lymfocyty imunologie mikrobiologie transplantace MeSH
- COVID-19 * komplikace mikrobiologie MeSH
- homologní transplantace MeSH
- kohortové studie MeSH
- lidé MeSH
- nemoc štěpu proti hostiteli etiologie imunologie MeSH
- rizikové faktory MeSH
- SARS-CoV-2 patogenita MeSH
- transplantace hematopoetických kmenových buněk * MeSH
- vakcíny proti COVID-19 škodlivé účinky MeSH
- Check Tag
- lidé MeSH
BACKGROUND: Allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients are at high risk of complications associated with COVID-19 infection due to dysfunction of their immune system. Vaccination can protect from the adverse consequences of COVID-19. However, studies on the efficacy of COVID-19 vaccines in HSCT recipients with insufficient post-HSCT immune reconstitution are still scarce. In our study, we determined how immunosuppressive medication and the reconstitution of the cellular immune system influenced T cell responses specific for the surface glycoprotein of SARS-CoV-2 virus (S antigen) after two doses of mRNA vaccine against COVID-19 in patients with myeloid malignancies treated with HSCT. METHODS: Vaccination outcomes were followed in 18 (allo-HSCT) recipients and 8 healthy volunteers. The IgG antibodies against SARS-CoV-2 spike (S) and nucleocapsid (NCP) protein were determined in ELISA and S-specific T cells were detected using a sensitive ELISPOT-IFNγ based on in vitro expansion and restimulation of T cells in pre- and post-vaccination blood samples. Multiparametric flow cytometry analysis of peripheral blood leukocyte differentiation markers was employed for determination of reconstitution of the main subpopulations of T cells and NK cells at month 6 after HSCT. RESULTS: S- specific IgG antibody response detected in 72% of the patients was lower than in healthy vaccinees (100%). Vaccine-induced T-cell responses to S1 or S2 antigen were significantly reduced in HSCT recipients, which were treated with corticosteroids in dose 5 mg of prednisone- equivalents or higher during the vaccination period or in preceeding 100 days in comparison with recipients un-affected with corticosteroids. A significant positive correlation was found between the level of anti-SARS-Cov-2 spike protein IgG antibodies and the number of functional S antigen-specific T cells. Further analysis also showed that the specific response to vaccination was significantly influenced by the interval between administration of vaccine and transplantation. Vaccination outcomes were not related to age, sex, type of mRNA vaccine used, basic diagnosis, HLA match between HSC donor and recipient, and blood counts of lymphocytes, neutrophils, and monocytes at the time of vaccination. Multiparametric flow cytometry analysis of peripheral blood leukocyte differentiation markers showed that good humoral and cellular S-specific immune responses induced by vaccination were associated with well-reconstituted CD4+ T cells, mainly CD4+ effector memory subpopulation at six months after HSCT. CONCLUSIONS: The results showed that both humoral and cellular adaptive immune responses of HSCT recipients to the SARS-CoV-2 vaccine were significantly suppressed by corticosteroid therapy. Specific response to the vaccine was significantly affected by the length of the interval between HSCT and vaccination. Vaccination as early as 5 months after HSCT can lead to a good response. Immune response to the vaccine is not related to age, gender, HLA match between HSC donor and recipient, or type of myeloid malignancy. Vaccine efficacy was dependent on well-reconstituted CD4+ T cells, at six months after HSCT.
- MeSH
- COVID-19 * prevence a kontrola MeSH
- imunita MeSH
- imunoglobulin G MeSH
- imunosupresivní léčba MeSH
- lidé MeSH
- mRNA vakcíny MeSH
- nádory * MeSH
- SARS-CoV-2 MeSH
- transplantace hematopoetických kmenových buněk * škodlivé účinky MeSH
- vakcíny proti COVID-19 MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Nucleophosmin (NPM1, B23) is a multifunctional phosphoprotein expressed in all tissues. The protein is mainly localized in nucleoli. In hematological malignancies, NPM1 belongs to commonly altered genes. Its mutation, always heterozygous, leads to the re-localization of the NPM1 protein from the nucleolus to the cytoplasm (NPM1c+). NPM1c+ is found in 30% of acute myeloid leukemia (AML). Our study showed that an AML patient, whose leukemia cells carried the NPM1c+ mutation and who was the recipient of allogeneic HSCT from a haploidentical donor, raised a robust allorestricted CD8+ T cell response directed against the NPM1wt protein. Favourably, the response against NPM1wt was not accompanied by side effects such as GvHD. Moreover, the induction of a high NPM1wt-specific response coincided with the decrease in NPM1c+ transcripts detected, implying a beneficial graft versus leukemia effect. On the basis of these results, we suppose that TCRs from allorestricted NPM1wt-specific T cells are worth studying in other recipients of grafts from haploidentical donors as a possible tool for TCR gene therapy.
- MeSH
- akutní myeloidní leukemie * genetika metabolismus terapie MeSH
- CD8-pozitivní T-lymfocyty metabolismus patologie MeSH
- hematopoetické kmenové buňky metabolismus patologie MeSH
- jaderné proteiny genetika metabolismus MeSH
- lidé MeSH
- nukleofosmin * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BK polyomavirus (BKPyV) persists lifelong in renal and urothelial cells with asymptomatic urinary shedding in healthy individuals. In some immunocompromised persons after transplantation of hematopoietic stem cells (HSCT), the BKPyV high-rate replication is associated with haemorrhagic cystitis (HC). We tested whether the status of BKPyV immunity prior to HSCT could provide evidence for the BKPyV tendency to reactivate. We have shown that measurement of pretransplant anti-BKPyV 1 and 4 IgG levels can be used to evaluate the HC risk. Patients with anti-BKPyV IgG in the range of the 1st-2nd quartile of positive values and with positive clinical risk markers have a significantly increased HC risk, in comparison to the reference group of patients with "non-reactive" anti-BKPyV IgG levels and with low clinical risk (LCR) (p = 0.0009). The predictive value of pretransplant BKPyV-specific IgG was confirmed by determination of genotypes of the shed virus. A positive predictive value was also found for pretransplant T-cell immunity to the BKPyV antigen VP1 because the magnitude of IFN-γ T-cell response inversely correlated with posttransplant DNAuria and with HC. Our novel data suggest that specific T-cells control BKPyV latency before HSCT, and in this way may influence BKPyV reactivation after HSCT. Our study has shown that prediction using a combination of clinical and immunological pretransplant risk factors can help early identification of HSCT recipients at high risk of BKPyV disease.
- Publikační typ
- časopisecké články MeSH
Adoptive transfer of multivirus-specific T cell lines (MVST) is an advanced tool for immunotherapy of virus infections after hematopoietic stem cell transplantation (HSCT). Their preparation includes activation of donor virus-specific T cells by the mixture of oligopeptides derived from immunodominant antigens of several most harmful viruses, i.e. human cytomegalovirus (HCMV), polyomavirus BK (BKV), Epstein-Barr virus (EBV) and adenovirus (ADV). The aim of our study was to find out whether antigenic competition may have an impact on the expansion of virus-specific T cells. MVST from several heathy blood donors were generated using a pulse of overlapping oligopeptides (PepMixes™, derived from the IE1 and pp65 CMV antigens, VP1 and LTAG BKV antigens, BZLF1 and EBNA1 proteins of EBV and hexon protein from ADV) and short time culture in the presence of IL-7 and IL-4. The amount of virus-specific T cells in MVST was measured by ELISPOT and flow cytometry after re-stimulation with individual antigens. To evaluate antigenic competition, MVST were expanded either with a complete set of antigens or with the mixture lacking some of them. MVST expanded with the antigen mixture including CMV antigens contained a lower proportion of the T cells of other antigen specificities. A similar inhibitory effect was not apparent for EBV-derived peptides. The competitive effect of CMV antigens was most pronounced in MVST from CMV-seropositive donors and was mediated by both IE1 and pp65-derived peptides. Antigenic competition did not influence the phenotype of either CMV- or BKV-specific T cells. Both T cell populations had an effector memory phenotype (CD45RO+, CD27-, CCR7-). However, CMV-specific T cells preferentially consist of CD8+ while in BKV-specific T cells, the CD4+ phenotype predominated. Modification of the MVST manufacture protocol to prevent antigenic competition may increase the efficacy of MVST in therapy of BKV infections in HSCT recipients.
- MeSH
- Adenoviridae imunologie patogenita MeSH
- adenovirové infekce lidí imunologie terapie virologie MeSH
- aktivace lymfocytů MeSH
- antigeny virové imunologie MeSH
- cytomegalovirové infekce imunologie terapie virologie MeSH
- Cytomegalovirus imunologie patogenita MeSH
- fenotyp MeSH
- imunodominantní epitopy MeSH
- imunoterapie adoptivní * MeSH
- infekce onkogenními viry imunologie terapie virologie MeSH
- infekce virem Epsteina-Barrové imunologie terapie virologie MeSH
- interakce hostitele a patogenu MeSH
- kultivované buňky MeSH
- lidé MeSH
- polyomavirové infekce imunologie terapie virologie MeSH
- T-lymfocyty imunologie transplantace virologie MeSH
- transplantace hematopoetických kmenových buněk škodlivé účinky MeSH
- virové nemoci imunologie terapie virologie MeSH
- virus BK imunologie patogenita MeSH
- virus Epsteinův-Barrové imunologie patogenita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
1 svazek : ilustrace, tabulky ; 30 cm
Recently, novel small DNA viruses (Human Polyomavirus HPyV6, HPyV7, HPyV9, Human Bocavirus HBoV1, Anelloviruses TTMV and TTMDV) have been discovered. Although these viruses belong to different family, they are widespread in population and they all presumably cause common lifelong infections. There is an increasing consideration that novel small DNA viruses play the key role as emerging opportunistic pathogens after reactivation in immunocompromised patients. The aim of the project is to gain knowledge of prevalence and biology of the newly discovered DNA viruses in healthy population and in immunocompromised individuals. The proposed study includes detection of viral DNA, determination of viral load, searching the persistence of the viruses and the presence of virus specific antibodies. In immunocompromised patient we will evaluate the possible clinical manifestation of viral infections.
Během posledních let byla objevena řada nových malých DNA virů (lidské polyomaviry HPyV6, HPyV7, HPyV9, lidský bocavirus HBoV1, anelloviry TTMV a TTMDV). Přestože tyto viry nepatří do jedné skupiny, všechny se pravděpodobně vyskytují běžně v lidské populaci, způsobují celoživotní perzistentní infekci organismu. K reaktivaci s možnými patologickými následky pak může docházet především u imunokompromitovaných jedinců. V tomto projektu se zaměříme na detekci a sledování perzistence nově objevených DNA virů u zdravých a imunokompromitovaných jedinců. Cílem studie je zjistit virovou nálož, variabilitu genomů a přítomnost virově specifických protilátek. U imunokompromitivaných jedinců budeme analyzovat klinické projevy, které mohou souviset s virovou infekcí.
- MeSH
- Anelloviridae MeSH
- DNA virů MeSH
- ELISA MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidský bocavirus MeSH
- polyomavirové infekce MeSH
- protilátky virové MeSH
- transplantace hematopoetických kmenových buněk MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- hematologie a transfuzní lékařství
- virologie
- biologie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu IGA MZ ČR
BACKGROUND: The reactivation of human cytomegalovirus (HCMV) in immunosuppressed patients is associated with significant morbidity. Testing HCMV-specific T-cell responses can help determine which patients are at high risk of HCMV disease. We optimized selection of HCMV antigens for detection of T-cell response of patients after allogeneic hematopoietic stem cell transplantation (HSCT) with the aim of identifying patients with insufficient control of HCMV reactivation. METHODS: T-cell immune response to HCMV was monitored in 30 patients during the first year after HSCT. The HSCT recipients were classified according to their anti-HCMV T-cell response and the presence of HCMV DNA in the blood. RESULTS: We observed an inverse relationship between the magnitude of HCMV-specific T-cell responses against CMV lysate, phosphoprotein (pp) 65, immediate early-1 (IE-1), UL36, and UL55, but not to US3 and US29 detected by interferon-gamma (IFNγ)- ELISPOT and the level of HCMV DNA in the blood of patients during the 30 days following sampling. The study has revealed that patients who received a graft from a seronegative donor have a lower T-cell response against HCMV and increased probability of HCMV reactivation in comparison to the patients who had received their graft from a seropositive donor. CONCLUSION: The individual peptide pools and native HCMV antigens were useful for monitoring the time course of the anti-HCMV response by IFNγ-ELISPOT, which proved to have a prognostic value. Besides widely employed peptide pools of pp65 and IE-1, the use of antigens UL36 and UL55, but not US3 or US29, increased sensitivity of the test.
- MeSH
- antigeny virové genetika imunologie MeSH
- antivirové látky terapeutické užití MeSH
- CD8-pozitivní T-lymfocyty imunologie virologie MeSH
- cytomegalovirové infekce farmakoterapie imunologie virologie MeSH
- Cytomegalovirus genetika imunologie MeSH
- ELISPOT MeSH
- fosfoproteiny imunologie MeSH
- imunokompromitovaný pacient MeSH
- interferon gama krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- transplantace hematopoetických kmenových buněk škodlivé účinky MeSH
- virové proteiny genetika imunologie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články 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.
Viral CC chemokine inhibitor (vCCI) of the clone P13 vaccinia virus (VACV) strain PRAHA lacks eight amino acids in the signal peptide sequence. To study the influence of vCCI on virus biology, a virus with the vCCI gene coding for a prolonged signal sequence was prepared. We found that secreted vCCI attenuated the virus in vivo, and that it correlated with decreased levels of RANTES, eotaxin, TARC, and MDC in the blood in comparison with the parental virus. We determined the influence of vCCI on the CTL response against VACV E3((140-148)) (VGPSNSPTF) and HPV16 E7((49-57)) (RAHYNIVTF) H-2D(b)-restricted epitopes. The examination of the specific CTL response elicited by immunization with the recombinant VACV-expressing tumor-associated HPV16 E7 antigen by IFN-γ ELISPOT showed that the immunogenicity of the recombinant VACV-producing secretory vCCI was similar to that of the parent virus or deletion mutant in the C23L/B29R locus. Immunization with the secretory vCCI-producing recombinant virus has a lower therapeutic anti-tumor effect against TC-1 tumors. Viral CCI downregulated the E7-specific response induced by gene gun immunization with the DNA vaccines pBSC-SigE7 LAMP and pBSC-vCCI. We also observed that the immune response against vCCI elicited by the DNA vaccine did not affect the multiplication of VACV in vivo.
- MeSH
- buněčné linie MeSH
- chemokin CCL17 krev MeSH
- chemokin CCL5 krev MeSH
- chemokiny CC antagonisté a inhibitory krev MeSH
- cytotoxické T-lymfocyty imunologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové supresorové proteiny krev MeSH
- Papillomavirus E7 - proteiny genetika imunologie MeSH
- proteiny ADAM krev MeSH
- protinádorové vakcíny genetika imunologie MeSH
- sekvenční delece MeSH
- syntetické vakcíny genetika imunologie MeSH
- vakcinace MeSH
- virové proteiny genetika metabolismus MeSH
- virové vakcíny imunologie MeSH
- virus vakcinie genetika imunologie patogenita MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The expression of the transcription factor encoded by the Wilms tumor gene 1 (WT1) is associated with a variety of human cancers. WT1 protein has been reported to serve as a target antigen for tumor-specific immune responses. We observed that the immunization of mice with peptide vaccines derived from WT1 in a mixture with the CpG adjuvant (ODN 1826) by tattoo administration was superior to subcutaneous delivery of the peptides in combination with CpG formulated with the mineral oil adjuvant or a DNA vaccine or a recombinant vaccinia virus vaccine expressing the truncated WT1 protein. Tattooing with the WT1122-140 and WT1126-134 peptide elicited the response of WT1-specific interferon-γ-producing T cells. Peptide vaccine administered with a tattoo device had an antitumor effect on the growth of the prostate tumor cell line TRAMP-C2, provided that the transforming growth factor-β produced by tumor cells was neutralized by anti-TGFβ monoclonal antibody. The treatment of the tumor-bearing mice with 5-azadeoxycytidine or poly IC did not work in synergy with the peptide vaccine.
- MeSH
- adjuvancia imunologická MeSH
- azacytidin analogy a deriváty farmakologie terapeutické užití MeSH
- injekce intradermální MeSH
- monoklonální protilátky imunologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory prostaty terapie MeSH
- poly I-C farmakologie terapeutické užití MeSH
- proteiny WT1 aplikace a dávkování imunologie MeSH
- protinádorové látky farmakologie terapeutické užití MeSH
- protinádorové vakcíny aplikace a dávkování MeSH
- subjednotkové vakcíny aplikace a dávkování MeSH
- tetování MeSH
- transformující růstový faktor beta antagonisté a inhibitory imunologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
