RESUMEN
OBJECTIVES: T cell immunity is key for the control of viral infections including SARS-CoV-2, in particular with regard to immune memory and protection against arising genetic variants. METHODS: We recently evaluated a peptide-based SARS-CoV-2 T cell activator termed CoVac-1 in a first-in-human trial in healthy adults. Here, we report on long-term safety and efficacy data of CoVac-1 until month 12. RESULTS: CoVac-1 is well tolerated without long-term immune-related side effects and induces long-lasting anti-viral T cell responses in 100% of study participants, with potent expandability of clusters of differentiation (CD4+) and CD8+ T cells targeting multiple different CoVac-1 T cell epitopes. T cell responses were associated with stronger injection site reaction. Beyond induction of T cell immunity, 89% of subjects developed CoVac-1-specific immunoglobulin G antibodies which associated with the intensity of the T cell response, indicating that CoVac-1-specific CD4+ T cells support the induction of B-cell responses. Vaccination with approved COVID-19 vaccines boosted CoVac-1-specific T cell responses. Overall, a low SARS-CoV-2 infection rate (8.3%) was observed. CONCLUSION: Together, a single application of CoVac-1 elicits long-lived and broad SARS-CoV-2-specific T cell immunity, which further supports the current evaluation of our T cell activator in patients with congenital or acquired B-cell defects.
Asunto(s)
COVID-19 , Adulto , Humanos , COVID-19/prevención & control , Vacunas contra la COVID-19 , Linfocitos T CD8-positivos , SARS-CoV-2 , Péptidos , Anticuerpos AntiviralesRESUMEN
T-cell immunity is central for control of COVID-19, particularly in patients incapable of mounting antibody responses. CoVac-1 is a peptide-based T-cell activator composed of SARS-CoV-2 epitopes with documented favorable safety profile and efficacy in terms of SARS-CoV-2-specific T-cell response. We here report a Phase I/II open-label trial (NCT04954469) in 54 patients with congenital or acquired B-cell deficiency receiving one subcutaneous CoVac-1 dose. Immunogenicity in terms of CoVac-1-induced T-cell responses and safety are the primary and secondary endpoints, respectively. No serious or grade 4 CoVac-1-related adverse events have been observed. Expected local granuloma formation has been observed in 94% of study subjects, whereas systemic reactogenicity has been mild or absent. SARS-CoV-2-specific T-cell responses have been induced in 86% of patients and are directed to multiple CoVac-1 peptides, not affected by any current Omicron variants and mediated by multifunctional T-helper 1 CD4+ T cells. CoVac-1-induced T-cell responses have exceeded those directed to the spike protein after mRNA-based vaccination of B-cell deficient patients and immunocompetent COVID-19 convalescents with and without seroconversion. Overall, our data show that CoVac-1 induces broad and potent T-cell responses in patients with B-cell/antibody deficiency with a favorable safety profile, which warrants advancement to pivotal Phase III safety and efficacy evaluation. ClinicalTrials.gov identifier NCT04954469.
Asunto(s)
Agammaglobulinemia , COVID-19 , Humanos , SARS-CoV-2 , Linfocitos T , Péptidos/uso terapéuticoRESUMEN
The DNAJB1-PRKACA fusion transcript is the oncogenic driver in fibrolamellar hepatocellular carcinoma, a lethal disease lacking specific therapies. This study reports on the identification, characterization, and immunotherapeutic application of HLA-presented neoantigens specific for the DNAJB1-PRKACA fusion transcript in fibrolamellar hepatocellular carcinoma. DNAJB1-PRKACA-derived HLA class I and HLA class II ligands induce multifunctional cytotoxic CD8+ and T-helper 1 CD4+ T cells, and their cellular processing and presentation in DNAJB1-PRKACA expressing tumor cells is demonstrated by mass spectrometry-based immunopeptidome analysis. Single-cell RNA sequencing further identifies multiple T cell receptors from DNAJB1-PRKACA-specific T cells. Vaccination of a fibrolamellar hepatocellular carcinoma patient, suffering from recurrent short interval disease relapses, with DNAJB1-PRKACA-derived peptides under continued Poly (ADP-ribose) polymerase inhibitor therapy induces multifunctional CD4+ T cells, with an activated T-helper 1 phenotype and high T cell receptor clonality. Vaccine-induced DNAJB1-PRKACA-specific T cell responses persist over time and, in contrast to various previous treatments, are accompanied by durable relapse free survival of the patient for more than 21 months post vaccination. Our preclinical and clinical findings identify the DNAJB1-PRKACA protein as source for immunogenic neoepitopes and corresponding T cell receptors and provide efficacy in a single-patient study of T cell-based immunotherapy specifically targeting this oncogenic fusion.
Asunto(s)
Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/terapia , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/terapia , Neoplasias Hepáticas/patología , Regulación Neoplásica de la Expresión Génica , Recurrencia Local de Neoplasia/genética , Proteínas de Fusión Oncogénica/genética , Inmunoterapia , Péptidos/genética , Proteínas del Choque Térmico HSP40/genética , Subunidades Catalíticas de Proteína Quinasa Dependientes de AMP CíclicoRESUMEN
T cell immunity is central for the control of viral infections. CoVac-1 is a peptide-based vaccine candidate, composed of SARS-CoV-2 T cell epitopes derived from various viral proteins1,2, combined with the Toll-like receptor 1/2 agonist XS15 emulsified in Montanide ISA51 VG, aiming to induce profound SARS-CoV-2 T cell immunity to combat COVID-19. Here we conducted a phase I open-label trial, recruiting 36 participants aged 18-80 years, who received a single subcutaneous CoVac-1 vaccination. The primary end point was safety analysed until day 56. Immunogenicity in terms of CoVac-1-induced T cell response was analysed as the main secondary end point until day 28 and in the follow-up until month 3. No serious adverse events and no grade 4 adverse events were observed. Expected local granuloma formation was observed in all study participants, whereas systemic reactogenicity was absent or mild. SARS-CoV-2-specific T cell responses targeting multiple vaccine peptides were induced in all study participants, mediated by multifunctional T helper 1 CD4+ and CD8+ T cells. CoVac-1-induced IFNγ T cell responses persisted in the follow-up analyses and surpassed those detected after SARS-CoV-2 infection as well as after vaccination with approved vaccines. Furthermore, vaccine-induced T cell responses were unaffected by current SARS-CoV-2 variants of concern. Together, CoVac-1 showed a favourable safety profile and induced broad, potent and variant of concern-independent T cell responses, supporting the presently ongoing evaluation in a phase II trial for patients with B cell or antibody deficiency.
Asunto(s)
Vacunas contra la COVID-19/inmunología , COVID-19/inmunología , SARS-CoV-2/inmunología , Linfocitos T/inmunología , Vacunas de Subunidad/inmunología , Administración Cutánea , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Linfocitos T CD8-positivos/inmunología , COVID-19/prevención & control , COVID-19/virología , Vacunas contra la COVID-19/administración & dosificación , Vacunas contra la COVID-19/efectos adversos , Ensayos Clínicos Fase II como Asunto , Femenino , Granuloma/inmunología , Humanos , Inmunogenicidad Vacunal , Interferón gamma/inmunología , Masculino , Persona de Mediana Edad , Linfocitos T Colaboradores-Inductores/inmunología , Vacunas de Subunidad/administración & dosificación , Vacunas de Subunidad/efectos adversos , Adulto JovenRESUMEN
BACKGROUND: A retrospective study using PET/CT imaging with 124I-labeled metaiodobenzylguanidine (124I-MIBG) was performed to estimate the (radiation) absorbed dose to the salivary glands in neuroendocrine cancer patients undergoing 131I-MIBG therapy and to compare these results with those in radioiodine (131I-iodide) therapy. METHODS: Twenty-seven patients received individual 124I-MIBG-PET/CT dosimetries, among whom 18 had not previously undergone any MIBG therapies (patient group before treatment) and 9 had already received MIBG therapies prior to the tracer dosimetries (patient group after treatment). For each patient, three or four 124I-MIBG PET/CT scans were performed at approximately 4 and 24 hours, as well as at approximately 48 or/and ≥96 hours after tracer injection. The absorbed doses per administered 131I-MIBG activity to the submandibular and parotid glands were calculated based on the MIRD concept, with its assumption of a uniform glandular activity distribution. RESULTS: The mean±standard deviation of the (self-)absorbed dose per activity averaged over both patient groups and salivary gland types was 0.53±0.24 Gy/GBq (median, 0.49 Gy/GBq; range, 0.17-1.38 Gy/GBq). The absorbed doses per activity of the patient group before treatment did not significantly deviate from those of the patient group after treatment (P=0.67). In the patient group after treatment, the mean±standard deviation of the cumulative 131I-MIBG activity was 20±12 GBq (median, 16 GBq; range, 10-50 GBq). Among the patient groups, no significant absorbed dose difference was found between the submandibular and parotid glands (P>0.24). In comparison to radioiodine therapy, the estimated absorbed dose per activity in MIBG was significantly higher (P<0.001), on average twice as high, contradicting the relationship between the absorbed dose and clinical observation of glandular side effects. CONCLUSIONS: The discrepant salivary gland responses in MIBG and radioiodine therapies suggest a different radiotherapeutical distribution on microscopic scale within the glandular tissue and prove the clinical relevance of a microdosimetric analysis.
Asunto(s)
3-Yodobencilguanidina/efectos adversos , Radioisótopos de Yodo/efectos adversos , Glándulas Salivales/efectos de la radiación , 3-Yodobencilguanidina/uso terapéutico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , Carcinoma Neuroendocrino/diagnóstico por imagen , Carcinoma Neuroendocrino/radioterapia , Niño , Femenino , Humanos , Radioisótopos de Yodo/uso terapéutico , Masculino , Persona de Mediana Edad , Tomografía Computarizada por Tomografía de Emisión de Positrones , Dosis de Radiación , Estudios Retrospectivos , Glándulas Salivales/diagnóstico por imagen , Adulto JovenRESUMEN
BACKGROUND: The goal of this prospective study was to estimate the absorbed (radiation) doses to salivary glands in radioiodine therapy of thyroid cancer under chewing-gum stimulation using (124)I positron emission tomography (PET)/computed tomography (CT) imaging. METHODS: Duplex ultrasonography was conducted in three test persons for visual comparison of the glandular blood flow with three different stimulation types (no stimulation, chewing tasteless gum base, sucking on lemon slices). Ten patients with newly diagnosed differentiated thyroid cancer received (124)I PET/CT dosimetry after thyroidectomy and prior to radioiodine therapy. Patients underwent a series of three (124)I PET/CT scans (4, 24, and ≥96 h after administration of 23 MBq (124)I). They were instructed to chew gum base (tasteless) approximately 20 min after ingesting the (124)I-containing capsule in the course of the first day. Absorbed doses per administered (131)I activity to the salivary glands were calculated and compared with the previously published results of the lemon-juice stimulation and non-stimulation groups. RESULTS: The sonograms in the three test persons showed that glandular blood perfusion by lemon-juice stimulation was clearly increased compared with non-stimulation or chewing of gum base. The sonogram comparison between the chewing-gum stimulation and non-stimulation demonstrated a minor increase of blood flow for the gum base-stimulated salivary glands. The mean ± standard deviation of the absorbed dose per activity under chewing-gum stimulation for the submandibular and parotid glands (within parentheses) was 0.22 ± 0.09 Gy/GBq (0.22 ± 0.08 Gy/GBq). Compared with the absorbed doses of the non-stimulation group, 0.24 ± 0.08 Gy/GBq (0.21 ± 0.05 Gy/GBq), those of the chewing-gum stimulation group showed no significant change (P > 0.60), but the absorbed doses of the lemon-juice stimulation group, 0.35 ± 0.14 Gy/GBq (0.33 ± 0.09 Gy/GBq), were significantly higher (P < 0.04) than those of the chewing-gum stimulation group. CONCLUSIONS: The results suggest that salivary flow induced by chewing gum base does not cause a significant reduction of the salivary gland absorbed dose compared with that in the non-stimulation group. The increased blood flow appears to be a decisive factor causing the increased (131)I absorbed doses in the lemon-juice stimulation group.
RESUMEN
Two genes coding for proteins with a high degree of sequence similarity to glycerol-3-phosphate dehydrogenases have been isolated from the yeast Pichiajadinii. Fragments of the genes were PCR-amplified with degenerated primers from genomic DNA of P. jadinii. Clones containing the full-length genes PjGPDI and PjGPD2 were isolated by screening genomic libraries. DNA sequencing revealed open reading frames (ORFs) of 1182 bp and 1185 bp for PjGpdlp and PjGpd2p, respectively. In a complementation study PjGPD1 rescued the growth defect of a Saccharomyces cerevisiae Agpdl mutant strain under osmotic stress, while complementation by PjGPD2 is temperature sensitive. The sequences of the PjGPD1 and PjGPD2 ORFs have been submitted to the EMBL Nucleotide Sequence Database under Accession No. AJ632339 and AJ632340, the sequences of the corresponding genomic DNA fragments under Accession No. AJ632341 and AJ635370, respectively.
Asunto(s)
Genes Fúngicos/genética , Glicerolfosfato Deshidrogenasa/genética , Pichia/genética , Secuencia de Aminoácidos , Secuencia de Bases , Cartilla de ADN , Prueba de Complementación Genética , Datos de Secuencia Molecular , Pichia/enzimología , Análisis de Secuencia de ADNRESUMEN
Rubinstein-Taybi syndrome (RSTS) is a distinct dominant disorder characterized by short stature, typical face, broad angulated thumbs and halluces, and mental retardation. The RSTS can be caused by chromosomal microdeletions and molecular mutations in the CREBBP gene; however, relatively few mutations have been reported to date. Here, we aimed to determine the rate of point mutations and other small molecular lesions in true RSTS and possible mild variants, by using genomic DNA sequencing. A consecutive series of patients including 17 patients from our previous study was investigated. We identified 19 causative mutations of CREBBP in a total of 45 patients representing three different diagnostic groups: (a) 17 mutations in 30 patients with unequivocal RSTS (detection rate 56.6%), (b) two mutations in eight patients with features suggestive of RSTS ("moderate or incomplete RSTS", detection rate 25%), and (c) no mutation in seven patients with undiagnosed syndromes and isolated features of RSTS. In general, the mutations were distributed without hot spots and most were unique; however, three recurrent mutations (R370X, R1664H, and N1978S) were identified. Furthermore, we detected 15 different intragenic polymorphisms, including two non-synonymous coding polymorphisms, L551I and Q2208H. We report not only the highest detection rate (56.6%) of CREBBP mutations in patients with RSTS to date, but also the second missense mutation (N1978S) in a patient with moderate or incomplete RSTS. Previous studies have identified cytogenetic deletions in the CREBBP gene in eight to 12% of patients and very recently, Roelfsema et al. reported EP300 gene mutations in three of 92 (3.3%) patients with either true RSTS or different syndromes resembling RSTS. Our 56.6% detection rate of molecular mutations in CREBBP in patients with unequivocal RSTS supports the new concept that RSTS is a genetically heterogeneous disorder and furthermore, indicates that RSTS may be caused by gene/s other than CREBBP in up to 30% of cases.