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Frontiers in Immunology 2023Reliable and sensitive characterization assays are important determinants of the successful clinical translation of immunotherapies. For the assessment of cytolytic...
Reliable and sensitive characterization assays are important determinants of the successful clinical translation of immunotherapies. For the assessment of cytolytic potential, the chromium 51 (Cr) release assay has long been considered the gold standard for testing effector cells. However, attaining the approvals to access and use radioactive isotopes is becoming increasingly complex, while technical aspects [i.e. sensitivity, short (4-6 hours) assay duration] may lead to suboptimal performance. This has been the case with our ex vivo expanded, polyclonal (CD4+ and CD8+) multivirus-specific T cell (multiVST) lines, which recognize 5 difficult-to-treat viruses [Adenovirus (AdV), BK virus (BKV), cytomegalovirus (CMV), Epstein Barr virus (EBV), and human herpes virus 6 (HHV6)] and when administered to allogeneic hematopoietic stem cell (HCT) or solid organ transplant (SOT) recipients have been associated with clinical benefit. However, despite mediating potent antiviral effects , capturing cytotoxic potential has proven difficult in a traditional Cr release assay. Now, in addition to cytotoxicity surrogates, including CD107a and Granzyme B, we report on an alternative, vital dye -based, flow cytometric platform in which superior sensitivity and prolonged effector:target co-culture duration enabled the reliable detection of both CD4- and CD8-mediated cytolytic activity against viral targets without non-specific effects.
Topics: Humans; Epstein-Barr Virus Infections; Herpesvirus 4, Human; Adenoviridae; BK Virus; Cell- and Tissue-Based Therapy
PubMed: 38187380
DOI: 10.3389/fimmu.2023.1299512 -
Journal of Clinical Microbiology Apr 2024BK virus (BKV) infection or reactivation in immunocompromised individuals can lead to adverse health consequences including BKV-associated nephropathy (BKVAN) in kidney...
UNLABELLED
BK virus (BKV) infection or reactivation in immunocompromised individuals can lead to adverse health consequences including BKV-associated nephropathy (BKVAN) in kidney transplant patients and BKV-associated hemorrhagic cystitis (BKV-HC) in allogeneic hematopoietic stem cell transplant recipients. Monitoring BKV viral load plays an important role in post-transplant patient care. This study evaluates the performance of the Alinity m BKV Investigational Use Only (IUO) assay. The linearity of the Alinity m BKV IUO assay had a correlation coefficient of 1.000 and precision of SD ≤ 0.25 Log IU/mL for all panel members tested (2.0-7.3 Log IU/mL). Detection rate at 50 IU/mL was 100%. Clinical plasma specimens tested comparing Alinity m BKV IUO to ELITech MGB Alert BKV lab-developed test (LDT) on the Abbott 2000 platform using specimen extraction protocols for DNA or total nucleic acid (TNA) resulted in coefficient of correlation of 0.900 and 0.963, respectively, and mean bias of 0.03 and -0.54 Log IU/mL, respectively. Alinity m BKV IUO compared with Altona RealStar BKV and Roche cobas BKV assays demonstrated coefficient of correlation of 0.941 and 0.980, respectively, and mean bias of -0.47 and -0.31 Log IU/mL, respectively. Urine specimens tested on Alintiy m BKV IUO and ELITech BKV LDT using TNA specimen extraction had a coefficient of correlation of 0.917 and mean bias of 0.29 Log IU/mL. The Alinity m BKV IUO assay was performed with high precision across the dynamic range and correlated well with other available BKV assays.
IMPORTANCE
BK virus (BKV) in transplant patients can lead to adverse health consequences. Viral load monitoring is important in post-transplant patient care. This study evaluates the Alinity m BKV assay with currently available assays.
Topics: Humans; BK Virus; Kidney Transplantation; Polyomavirus Infections; Viral Load; Nucleic Acids; Tumor Virus Infections
PubMed: 38526061
DOI: 10.1128/jcm.01354-23 -
Journal of Travel Medicine Mar 2024The global spread of the chikungunya virus (CHIKV) increases the exposure risk for individuals travelling to or living in endemic areas. This Phase 3 study was designed... (Randomized Controlled Trial)
Randomized Controlled Trial
A randomized, double-blinded Phase 3 study to demonstrate lot-to-lot consistency and to confirm immunogenicity and safety of the live-attenuated chikungunya virus vaccine candidate VLA1553 in healthy adults†.
BACKGROUND
The global spread of the chikungunya virus (CHIKV) increases the exposure risk for individuals travelling to or living in endemic areas. This Phase 3 study was designed to demonstrate manufacturing consistency between three lots of the single shot live-attenuated CHIKV vaccine VLA1553, and to confirm the promising immunogenicity and safety data obtained in previous trials.
METHODS
This randomized, double-blinded, lot-to-lot consistency, Phase 3 study, assessed immunogenicity and safety of VLA1553 in 408 healthy adults (18-45 years) in 12 sites across the USA. The primary endpoint was a comparison of the geometric mean titre (GMT) ratios of CHIKV-specific neutralizing antibodies between three VLA1553 lots at 28 days post-vaccination. Secondary endpoints included immunogenicity and safety over 6 months post-vaccination.
RESULTS
GMTs were comparable between the lots meeting the acceptance criteria for equivalence. The average GMT (measured by 50% CHIKV micro plaque neutralization test; μPRNT50) peaked with 2643 at 28 days post-vaccination and decreased to 709 at 6 months post-vaccination. An excellent seroresponse rate (defined as μPRNT50 titre ≥ 150 considered protective) was achieved in 97.8% of participants at 28 days post-vaccination and still persisted in 96% at 6 months after vaccination. Upon VLA1553 immunization, 72.5% of participants experienced adverse events (AEs), without significant differences between lots (related solicited systemic AE: 53.9% of participants; related solicited local AE: 19.4%). Overall, AEs were mostly mild or moderate and resolved without sequela, usually within 3 days. With 3.9% of participants experiencing severe AEs, 2.7% were classified as related, whereas none of the six reported serious adverse events was related to the administration of VLA1553.
CONCLUSIONS
All three lots of VLA1553 recapitulated the safety and immunogenicity profiles of a preceding Phase 3 study, fulfilling pre-defined consistency requirements. These results highlight the manufacturability of VLA1553, a promising vaccine for the prevention of CHIKV disease for those living in or travelling to endemic areas.
Topics: Adult; Humans; Antibodies, Neutralizing; Antibodies, Viral; Chikungunya Fever; Chikungunya virus; Double-Blind Method; Neutralization Tests; Vaccines, Attenuated; Adolescent; Young Adult; Middle Aged
PubMed: 38091981
DOI: 10.1093/jtm/taad156 -
Antiviral Research Feb 2024JC polyomavirus (JCPyV) is a nonenveloped, double-stranded DNA virus that infects the majority of the population. Immunocompetent individuals harbor infection in their...
JC polyomavirus (JCPyV) is a nonenveloped, double-stranded DNA virus that infects the majority of the population. Immunocompetent individuals harbor infection in their kidneys, while severe immunosuppression can result in JCPyV spread to the brain, causing the neurodegenerative disease progressive multifocal leukoencephalopathy (PML). Due to a lack of approved therapies to treat JCPyV and PML, the disease results in rapid deterioration, and is often fatal. In order to identify potential antiviral treatments for JCPyV, a high-throughput, large-scale drug screen was performed using the National Institutes of Health Clinical Collection (NCC). Drugs from the NCC were tested for inhibitory effects on JCPyV infection, and drugs from various classes that reduced JCPyV infection were identified, including receptor agonists and antagonists, calcium signaling modulators, and enzyme inhibitors. Given the role of calcium signaling in viral infection including Merkel cell polyomavirus and simian virus 40 polyomavirus (SV40), calcium signaling inhibitors were further explored for the capacity to impact JCPyV infection. Calcium and calmodulin inhibitors trifluoperazine (TFP), W-7, tetrandrine, and nifedipine reduced JCPyV infection, and TFP specifically reduced viral internalization. Additionally, TFP and W-7 reduced infection by BK polyomavirus, SV40, and SARS-CoV-2. These results highlight specific inhibitors, some FDA-approved, for the possible treatment and prevention of JCPyV and several other viruses, and further illuminate the calcium and calmodulin pathway as a potential target for antiviral drug development.
Topics: Humans; Calcium; Calmodulin; Neurodegenerative Diseases; Leukoencephalopathy, Progressive Multifocal; Polyomavirus Infections; JC Virus; Simian virus 40; Antiviral Agents; Sulfonamides
PubMed: 38246207
DOI: 10.1016/j.antiviral.2024.105817 -
Pathogens (Basel, Switzerland) Apr 2024In the absence of an effective antiviral treatment for BK polyomavirus (BKPyV), a better understanding of the epidemiology and time course of BKPyV replication after...
BACKGROUND/OBJECTIVES
In the absence of an effective antiviral treatment for BK polyomavirus (BKPyV), a better understanding of the epidemiology and time course of BKPyV replication after kidney transplantation is needed to limit the virus's impact on the graft outcome.
METHODS
In a 7-year study, we screened more than 430 kidney transplant recipients and analyzed the time course and virological characteristics of BKPyV replication.
RESULTS
Urinary viral replication was observed in 116 (27%) of the 430 patients, and 90 of the 116 (78%) had viral DNAemia. Thirty-eight patients (8.8%) were presumed to have nephropathy (DNAemia > 4 log10 copies/mL). Of the patients with BKPyV replication, 48%, 60%, 71%, and 80% were first found to be positive one, two, three, and four months post-transplantation. The initial viral load in the urine was below 7 log10 copies/mL in 100% of the patients with viral replication first detected before the first month, and this proportion was 57% when viral replication was first detected after the first month. When the BKPyV replication was first detected in a urine sample at month 3 or later, 81.5% of patients had concomitant BKPyV DNAemia. The predominant viral subtype was Ib2 (60%), and there was no apparent relationship between the subtype and the time course of BKPyV replication.
CONCLUSIONS
Urinary BKPyV replication occurs early after renal transplantation and in most patients will increase to a level requiring therapeutic intervention. Close monitoring for BKPyV in the early post-transplantation period would enable the pre-emptive adjustment of the immunosuppression regimen.
PubMed: 38668270
DOI: 10.3390/pathogens13040315 -
Signal Transduction and Targeted Therapy May 2024Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a 'highly transmissible respiratory pathogen, leading to severe multi-organ damage. However, knowledge...
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a 'highly transmissible respiratory pathogen, leading to severe multi-organ damage. However, knowledge regarding SARS-CoV-2-induced cellular alterations is limited. In this study, we report that SARS-CoV-2 aberrantly elevates mitochondrial bioenergetics and activates the EGFR-mediated cell survival signal cascade during the early stage of viral infection. SARS-CoV-2 causes an increase in mitochondrial transmembrane potential via the SARS-CoV-2 RNA-nucleocapsid cluster, thereby abnormally promoting mitochondrial elongation and the OXPHOS process, followed by enhancing ATP production. Furthermore, SARS-CoV-2 activates the EGFR signal cascade and subsequently induces mitochondrial EGFR trafficking, contributing to abnormal OXPHOS process and viral propagation. Approved EGFR inhibitors remarkably reduce SARS-CoV-2 propagation, among which vandetanib exhibits the highest antiviral efficacy. Treatment of SARS-CoV-2-infected cells with vandetanib decreases SARS-CoV-2-induced EGFR trafficking to the mitochondria and restores SARS-CoV-2-induced aberrant elevation in OXPHOS process and ATP generation, thereby resulting in the reduction of SARS-CoV-2 propagation. Furthermore, oral administration of vandetanib to SARS-CoV-2-infected hACE2 transgenic mice reduces SARS-CoV-2 propagation in lung tissue and mitigates SARS-CoV-2-induced lung inflammation. Vandetanib also exhibits potent antiviral activity against various SARS-CoV-2 variants of concern, including alpha, beta, delta and omicron, in in vitro cell culture experiments. Taken together, our findings provide novel insight into SARS-CoV-2-induced alterations in mitochondrial dynamics and EGFR trafficking during the early stage of viral infection and their roles in robust SARS-CoV-2 propagation, suggesting that EGFR is an attractive host target for combating COVID-19.
Topics: SARS-CoV-2; Mitochondria; Humans; Animals; Mice; COVID-19; ErbB Receptors; Virus Replication; Energy Metabolism; Vero Cells; Chlorocebus aethiops; Antiviral Agents; COVID-19 Drug Treatment; Membrane Potential, Mitochondrial; Oxidative Phosphorylation; Signal Transduction
PubMed: 38734691
DOI: 10.1038/s41392-024-01836-x -
Virology Journal Jul 2023Human polyomavirus BK (BKPyV) causes associated nephropathy and contributes to urinary tract cancer development in renal transplant recipients. Large tumor antigen (LT)...
BACKGROUND
Human polyomavirus BK (BKPyV) causes associated nephropathy and contributes to urinary tract cancer development in renal transplant recipients. Large tumor antigen (LT) is an early protein essential in the polyomavirus life cycle. Protein acetylation plays a critical role in regulating protein stability, so this study investigated the acetylation of the BKPyV LT protein.
METHODS
The BKPyV LT nucleotide was synthesized, and the protein was expressed by transfection into permissive cells. The BKPyV LT protein was immunoprecipitated and subjected to LC-MS/MS analysis to determine the acetylation residues. The relative lysine was then mutated to arginine in the LT nucleotide and BKPyV genome to analyze the role of LT lysine acetylation in the BKPyV life cycle.
RESULTS
BKPyV LT acetylation sites were identified at Lys3 and Lys230 by mass spectrometry. HDAC3 and HDAC8 and their deacetylation activity are required for BKPyV LT expression. In addition, mutations of Lys3 and Lys230 to arginine increased LT expression, and the interaction of HDAC3 and LT was confirmed by coimmunoprecipitation.
CONCLUSIONS
HDAC3 is a newly identified protein that interacts with BKPyV LT, and LT acetylation plays a vital role in the BKPyV life cycle.
Topics: Humans; Polyomavirus; BK Virus; Kidney Transplantation; Lysine; Chromatography, Liquid; Polyomavirus Infections; Tandem Mass Spectrometry; Antigens, Neoplasm; Protein Stability; Tumor Virus Infections; Histone Deacetylases; Repressor Proteins
PubMed: 37464367
DOI: 10.1186/s12985-023-02128-6 -
Chirurgie (Heidelberg, Germany) Aug 2023The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic resulted in many infections with the virus and sickness due to coronavirus disease 2019... (Review)
Review
[COVID-19 as an insurance case of the statutory accident insurance: occupational disease or occupational accident : Relevant knowledge for the (general and abdominal) surgeon].
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic resulted in many infections with the virus and sickness due to coronavirus disease 2019 (COVID-19). Therefore, there was a dramatic increase in the number of reported and recognized occupational diseases (Berufskrankheit, BK) and occupational accidents (Arbeitsunfall, AU) at the German Social Accident Insurance Institutions (BG) and accident insurance funds (Unfallkassen).The publication aims to show the differences between BK and AU and to review the current data on occupational diseases. It deals with the definitions of BK and AU, the differences in the conditions for recognition as BK or AU. Furthermore, the claims for benefits are presented. Finally, statistical key figures of the BK according to No. 3101 and the AU are presented.Results (key points)- According to § 7 SGB VII, AU and BK are insured events of the statutory accident insurance.- In surgery, like specifically in the rest of the healthcare system, the relevance of the SARS-CoV‑2 infection with post-COVID in personnel for occupational medical prevention and as a case to be recognized by the statutory accident insurance (BK or AU) becomes clear.- Relevant for the recognition are the duration and the intensity of the contact (local proximity) and the SARS-CoV‑2 occupational health and safety rule of 20 August 2020 essentially recognizes a contact duration of at least 15 min at a spatial distance of less than 1.5-2 m (further aspects: more intensive shorter contacts, number of verifiably infected persons in the closer activity environment or the usual personal contacts, spatial situation, work route, special constellations).- No case numbers can be elicited for the detailed presentation of the surgery setting.- There are still immense problems and challenges in the assessment of COVID-19 consequences or post-COVID as occupational disease, as numerous uncertainty factors, such as insufficiently secured knowledge about the further long-term course over the years or the widely varied symptom spectrum complicates the medical assessment of the consequences of this disease.Conclusion: the SARS-CoV‑2 pandemic is a special challenge for surgery with intensive patient contact and for the entire healthcare system. This caused long-lasting changes and the adequate health care as well as insurance law processing of the (case-specific) consequences might still require considerable efforts and resources.
Topics: Humans; COVID-19; Accidents, Occupational; Insurance, Accident; SARS-CoV-2; Occupational Diseases; Surgeons
PubMed: 37266705
DOI: 10.1007/s00104-023-01892-z -
International Journal of Surgical... Dec 2023We report a 32-year-old male 14 years post-living-related kidney transplant presenting with new-onset hematuria and BK viremia. He was found to have BK virus-associated...
We report a 32-year-old male 14 years post-living-related kidney transplant presenting with new-onset hematuria and BK viremia. He was found to have BK virus-associated urothelial carcinoma originating in the renal allograft with locally advanced disease and metastases to multiple sites. He also developed acute T-cell-mediated rejection in the setting of immunosuppression reduction for BK viremia prior to undergoing transplant nephrectomy. Eight months following transplant nephrectomy and immunosuppression cessation, distant metastases persisted with partial response to chemotherapy and immunotherapy. Here, we discuss this unique presentation and compare it with other BK virus-associated allograft carcinomas reported in the literature, in addition to discussing evidence for the role of BK virus in oncogenesis.
Topics: Male; Humans; Adult; Kidney Transplantation; Carcinoma, Transitional Cell; BK Virus; Viremia; Urinary Bladder Neoplasms; Allografts; Nephrectomy; Tumor Virus Infections
PubMed: 37013271
DOI: 10.1177/10668969231160258 -
Health Science Reports Apr 2024Gastric cancer is a significant global issue with a high death rate. This malignancy could be associated with several viral agents such as EBV, CMV, HHV-6, JCV, and BKV.
Investigation of Epstein-Barr virus, Cytomegalovirus, Human herpesvirus 6, and Polyoma viruses (JC virus, BK virus) among Gastric cancer patients: A cross sectional study.
BACKGROUND AND AIMS
Gastric cancer is a significant global issue with a high death rate. This malignancy could be associated with several viral agents such as EBV, CMV, HHV-6, JCV, and BKV.
OBJECTIVE
Evaluation of EBV, CMV, HHV-6 ,and JCV, BKV frequency among gastric cancer patients.
METHODS
In this cross-sectional study, a total number of 60 gastric cancer specimens (32 male, 28 female) were retrieved from the pathology lab. Formalin-fixed paraffin-embedded tissue was used for molecular testing. DNA was extracted from samples, according to protocol, and used for PCR reaction. Polymerase chain reactions were used to assess CMV, EBV, HHV-6, JCV, and BKV frequency.
RESULTS AND CONCLUSION
The mean age of the participants was 61 years and 53.3% (32) of the participants were Male. A total number of 5 samples (8.34%) were infected with viral agents. Four male gastric samples were infected with EBV (6.67%) and only one female sample contained the BKV genome (1.67%). Totally 8.34% of the samples were infected with EBV and BKV. The CMV, HHV-6, and JCV genome was not detected in the samples. In conclusion, the presence of two viral agents including EBV and BKV among male and female samples respectively, and the genome of other viruses were not detected.
PubMed: 38650724
DOI: 10.1002/hsr2.2043