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Background: Currently there are no published data regarding SARS-CoV-2 vaccine efficacy in paediatric-onset multiple sclerosis (MS) on disease-modifying treatments (DMTs). In adults, DMTs such as SIP-inhibitors (e.g., Fingolimod) and anti-CD20 therapies (e.g., Ocrelizumab) have been found to diminish SARS-CoV-2 vaccine efficacy. Objective(s): To ascertain whether young people with MS on DMTs generate a protective B-and/or T-cell response following SARS-CoV-2 vaccination or wild type (WT) infection. Method(s): Five millilitre additional blood was taken from MS patients during routine surveillance phlebotomy. Vaccination status and exposure to WT COVID was recorded. Serum samples measured SARS-CoV-2 antibodies using MSD quantitative assay. Multiplex T-cell stimulation assay was used to measure T-cell activity and T-cell response to SARS-CoV-2 Spike peptide was analysed. Result(s): Thirty-one MS patients (M:F 5:26;11-18 years) were included. DMTs included Ocrelizumab (n = 26);Fingolimod (n = 2);other DMT (n = 3). Vaccination status was confirmed in 12 children. 26/31 patients demonstrated protective B-cell responses, of which 10 were vaccinated +/- previous infection;six previously infected with unconfirmed vaccine status;and 10 had no infection/vaccine data available. Of the five patients with no B-cell response two were vaccinated. Six patients had impaired T-cell proliferation but 5/6 generated a B-cell response. T-cell proliferation upon exposure to Spike peptide was seen in 10 children'3/10 were vaccinated and 9/10 were being treated with Ocrelizumab. Patients on Fingolimod had impaired T-cell activity and no response to SARS-CoV-2 peptides, despite one being vaccinated and having a B-cell response. Conclusion(s): Fingolimod appeared to impair T-cell responses, however, only two patients were on this treatment. Those on Ocrelizumab have some protection following vaccination or WT exposure with 5/26 patients showing an impaired B-cell response but partial preservation of T-cell responses. While vaccination prior to starting DMT is ideal, some protection will be generated whilst on treatment.
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Cancer gene therapy based on various gene delivery vectors has some potential but also has obvious disadvantages. In this study, a new M13 phage-based oncolytic virus is constructed that carried the RGD peptides to target tumor cells and the 3C gene of Seneca Valley virus (SVV) preceded by a eukaryotic initial transcriptional region (ITR) to transcribe an oncolytic protein to kill tumor cells. Recombinant virus particles of 1200 nm in length are obtained in large quantities by transfecting the recombinant M13 phage plasmid into the host BL2738 and are investigated in vitro in tumor cells and in vivo in tumor-bearing mice to evaluate their antitumor effect. The experiments using Hela cells confirm that the engineered M13 phage can target and enter Hela cells, and express the SVV 3C protein, resulting in apoptosis of target cells by upregulating the expression of caspase 3. Furthermore, the results of experiments in vivo also show that the recombinant phage significantly inhibits the enhanced tumor volume in nude mice compared to the control groups. The M13 phage may be engineered to fuse with a variety of oncolytic proteins to inhibit the growth of tumor cells in the future, providing a promising phage-based targeted oncolytic reagent.
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BACKGROUND: Patients on hemodialysis (HD) are at high risk of a severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infection, which causes coronavirus disease 2019 (COVID-19), and humoral and cellular response data in these patients are limited. SARS-CoV-2 has four major structural proteins: spike (S), membrane, envelope and nucleocapsid (N) proteins. The S protein consists of the S1 and S2 subunits, which mediate cell surface binding via the receptor-binding domain, and induce viral-host cell membrane fusion, respectively. Approximately, 90% of the SARS-CoV-2-specific neutralizing antibodies (nAbs) target the highly immunogenic receptor-binding domain, but there is little evidence of nAbs targeting other viral structural proteins, such as the N protein. Speer et al. reported reduced Ab responses to the first and second doses of the mRNA vaccine, BNT162b2 (BioNTech), in patients on long-term HD. The majority (82%) of patients developed nAbs after the second dose, but at lower levels than healthy controls (HCs) [1]. Since data on the efficacy of COVID-19 vaccination in patients on HD are limited, we investigated the SARS-CoV-2 nAbs in Japanese patients on HD. METHOD: Forty-two patients (24 males and 18 females) on HD for an average of 4.5 years (0-21), with a mean age of 68 years (28-92), who received two doses of the mRNA vaccine, BNT162b2, between May 1 and 30 November 2021 were included. In addition, 10 HCs (6 males and 4 females with mean age of 55 (26-75) and 41 (28-63) years, respectively) with normal renal function who received two doses of the mRNA vaccine BNT162b2 in the same period were tested for SARS-CoV-2 nAbs at 4, 8, 12, 16 and 20 weeks (W) after vaccination. We measured SARS-CoV-2 Abs in human plasma qualitatively with a fully automated Cobas e801 analyzer, an Elecsys ® Anti-SARS-CoV-2 electrochemiluminescence immunoassay, and an Elecsys ® Anti-SARS-CoV-2 S RUO electrochemiluminescence immunoassay (Roche Diagnostics International Ltd, Rotkreuz, Switzerland) according to the manufacturer's instructions. RESULTS: Approximately, 97.6% (41/42) of patients on HD tested positive for SARSCoV-2 nAbs. The mean titer of SARS-CoV-2 nAbs after BNT162b2 vaccination in all individuals was 490.1 (0.4-5116) U/mL. The mean titers of SARS-CoV-2 nAbs after BNT162b2 vaccination in patients on HD aged between 56-92 years and 28-53 years were 266.7 (0.4-1131) U/mL and 1320.4 (44.8-5116) U/mL, respectively. The levels of nAbs were lower in the older group than in the younger group. Patients on HD are associated with premature aging of the immune systems. Progressive immunosenescence may be associated with reduced T cell activity and humoral response, potentially leading to reduced vaccine protection. During the follow-up period, Abs against N protein were not detected in all individuals. Over 90% of patients on HD wore face masks, washed their hands and maintained a 2 m social distance in the dialysis facilities. Almost all patients avoided the three Cs (closed spaces with poor ventilation, crowded places with nearby people and close-contact settings, such as close-range conversations). In contrast, the mean SARS-CoV-2 nAb titers in the older (55-73 years) HCs were 1006, 643, 520, 464 and 390 U/mL at 4, 8, 12, 16 and 20 W post-vaccination. Contrastingly, the mean SARS-CoV-2 nAb titers in the younger (26-45 years) HCs were 2685, 2032, 1731, 1609 and 1527 U/mL at 4, 8, 12, 16 and 20 W post-vaccination. The nAb levels decreased gradually and were lower in the older group than in the younger group. CONCLUSION: Japanese patients on HD had SARS-CoV-2 neutralizing capacities after BNT162b2 vaccination. The majority (97.6%) developed nAb after the second dose. The levels of neutralizing antibodies were lower in the older group than in the younger group.
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Background: Inactivated SARS-CoV-2 vaccine (CoronaVac) is commonly used in national immunization programmes. However, an immune response of CoronaVac significantly declined after 3 months, our study, thus, aimed to explore the immune response against COVID-19 following the booster dose by assessing both B-cell and T-cell activities compared to the convalescent samples. Methods: In this prospective cohort study, 98 healthcare workers with a 2-dose CoronaVac vaccination with a subsequent booster dose of ChAdOx1 nCoV-19 (n=56) or BNT162b2 (n=42) were included during March and October 2021. Immune responses were evaluated by surrogated viral neutralization test (sVNT, cPass™), anti-SARS-CoV-2 RBD total antibodies (Elecsys®) and the ELISPOT with spike (S1) peptide pools. The samples were analyzed at baseline, 4 and 12 weeks after the second CoronaVac and 4 weeks after a booster dose. In addition, convalescent sera and peripheral blood mononuclear cells (PBMCs) of the COVID-19 patients were collected at 4 weeks after diagnosis. Results: Median (interquartile range, IQR) age was 40 (31-52) years old with female predominant (80%). The median (IQR) interval after the second CoronaVac was 88 (74-92) days for ChAdOx1 nCoV-19 and 113 (112-115) days for BNT162b2. There was a significant decrease in neutralizing antibodies at the 12th week after primary CoronaVac vaccination (Figure 1). At 4 weeks after the ChAdOx1 nCoV-19 booster, median (IQR) level of sVNT and anti-RBD total antibody levels were 98.1% (97.9-98.2%) and 7768 (5349-11142), respectively, which were significantly different from the BNT162b2 booster, 98.5% (98.5-98.6%) and 25129 (17531-39434) BAU/mL, respectively (p<0.001 both). The antibody levels of the booster vaccine group were significantly higher than in the COVID-19 patients, which median (IQR) of sVNT was 80.8% (61.7-94.2%) in the mild COVID-19 and 93.8% (85.4-95.6%) in COVID-19 pneumonia, while anti-RBD total antibody levels were 94 (22-207) and 222 (130-378) BAU/mL, respectively. Using the ELISPOT with S1 peptide pools, median (IQR) of T cell response was 106 (24-256) and 196 (60-244) Spot Forming Unit (SFU)/millions of PBMCs for ChAdOx1 nCoV-19 and BNT162b2, respectively (p 0.49) which were comparable to the COVID-19 cases. Conclusion: A 2-dose CoronaVac followed by ChAdOx1 nCoV-19 or BNT162b2 effectively boosted a significantly higher antibody response than the natural COVID-19 infection. In addition, BNT162b2 booster induced significantly higher antibody levels than ChAdOx1 nCoV-19.
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Coronavirus disease 2019 (COVID-19) has spread rapidly and led to over 5 million deaths to date globally. Due to the successively emerging mutant strains, therapeutics and prevention against the causative virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are urgently needed. Prevention of SARS-CoV-2 infection in public and hospital areas is essential to reduce the frequency of infections. Silver nanoparticles (AgNPs) with virucidal effects have been reported. Therefore, we investigated the virucidal activity and safety of ten types of AgNPs with different surface modifications and particle sizes, in cells exposed to SARS-CoV-2 in vitro. The AgNPs could effectively inhibit the activity of SARS-CoV-2, and different surface modifications and particle sizes conferred different virucidal effects, of which 50-nm BPEI showed the strongest antiviral effect. We concluded that the efficacy of each type of AgNP type was positively correlated with the corresponding potential difference (R2 = 0.82). These in vitro experimental data provide scientific support for the development of therapeutics against COVID-19, as well as a research basis for the development of broad-spectrum virucides. Given the increasing acquired resistance of pathogens against conventional chemical and antibody-based drugs, AgNPs may well be a possible solution for cutting off the route of transmission, either as an external material or a potential medicine.
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Background. Global surveillance has identified emerging SARS-CoV-2 variants of concern (VOC) associated with increased transmissibility, disease severity, and resistance to neutralization by current vaccines under emergency use authorization (EUA). Here we assessed cross-immune responses of INO-4800 vaccinated subjects against SARS-CoV-2 VOCs. Methods. We used a SARS-CoV-2 IgG ELISA and a pseudo neutralization assay to assess humoral responses, and an IFNγ ELISpot to measure cellular responses against SARS-CoV-2 VOC in subjects immunized with the DNA vaccine, INO-4800. Results. IgG binding titers were not impacted between wild-type (WT) and B.1.1.7 or B.1.351 variants. An average 1.9-fold reduction was observed for the P.1 variant in subjects tested at week 8 after receiving two doses of INO-4800 (Figure 1a). We performed a SARS-CoV-2 pseudovirus neutralization assay using sera collected from 13 subjects two weeks after administration of a third dose of either 0.5 mg, 1 mg, or 2 mg of INO-4800. Neutralization was detected against WT and the emerging variants in all samples tested. The mean ID50 titers for the WT, B.1.1.7, B.1.351 and P.1. were 643 (range: 70-729), 295 (range: 46-886), 105 (range: 25-309), and 664 (range: 25-2087), respectively. Compared to WT, there was a 2.1 and 6.9-fold reduction for B.1.1.7 and B.1.351, respectively, while there was no difference between WT and the P.1 variant (Figure 1b). Next, we compared cellular immune responses to WT and SARS-CoV-2 Spike variants elicited by INO-4800 vaccination. We observed similar cellular responses to WT (median = 82.2 IQR = 58.9-205.3), B.1.1.7 (79.4, IQR = 38.9- 179.7), B.1.351 (80, IQR = 40.0-208.6) and P.1 (78.3, IQR = 53.1-177.8) Spike peptides (Figure 2). Conclusion. INO-4800 vaccination induced neutralizing antibodies against all variants tested, with reduced levels detected against B.1.351. IFNγ T cell responses were fully maintained against all variants tested.
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CD6 is a co-stimulatory receptor predominantly expressed on T cells that acts as a crucial regulator of T-cell activation and is implicated in the pathogenesis of multiple autoimmune diseases. Activated leukocyte cell adhesion molecule (ALCAM), a CD6 ligand, is expressed on antigen-presenting cells, as well as epithelial and endothelial cells of acute GVHD target organs, including the skin and the GI tract.1,2 Previous studies in patients receiving alloHCT showed that ex vivo depletion of donor CD6-positive T cells lowered the incidence of acute GVHD, providing a rationale for therapeutically targeting CD6 in acute GVHD.3 Itolizumab, a humanized immunoglobulin G1 monoclonal antibody undergoing evaluation as treatment for acute GVHD, binds CD6 and blocks interaction with ALCAM to inhibit T-cell activity and trafficking.4 Dr John Koreth presented interim study results from EQUATE, an ongoing phase 1b/2 study of itolizumab in combination with corticosteroids for newly diagnosed severe acute GVHD (grade 3-4) after first alloHCT.5 The phase 1b portion is an open-label, 3+3 dose-escalation study evaluating doses of 0.4, 0.8, 1.6, and 2.4 mg/kg administered intravenously every 2 weeks through day 57. As of November 13, 2020, 10 patients completed treatment: 4 with 0.4 mg/kg, 3 with 0.8 mg/kg, and 3 with 1.6 mg/kg. All patients received corticosteroids at an initial dose of 1 to 2 mg/kg/day. Their mean age was 48 years (standard deviation, 15.7 years), 90% were male, and 90% were white. The graft source was peripheral blood for 80% and bone marrow for 20%, and 80% had an HLA-matched donor. The mean time to onset of GVHD was 43 days, and 100% of patients had GI involvement. At day 57, the ORR was 50% with 0.4 mg/kg, 100% with 0.8 mg/kg, and 100% with 1.6 mg/kg.5 The median percent corticosteroid dose reduction at day 85 was 93%, 87%, and 91% for the 0.4, 0.8, and 1.6 mg/kg groups, respectively (Figure 6). Itolizumab decreased the CD6 levels on T cells in a dose-dependent manner within 24 hours of the first dose, a reduction that was maintained throughout the treatment period. Across the dosing cohorts, all patients developed at least 1 AE. Most AEs were mild to moderate in severity.5 The most common AEs were hypomagnesemia (n=3) and peripheral edema (n=3). One mild infusion reaction was noted. Six patients had serious AEs, including recurrent GVHD (n=1), sepsis (n=2;1 event was considered a dose-limiting toxicity), fever (n=1), COVID-19 (n=1), and disseminated nocardia (n=1).
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TYPE: TOPIC: Pharmacotherapeutics PURPOSE: The i-NEB-MiniTM is a jet nebulizer designed for controlled delivery of small (< 2 mL) volumes of potent drugs to the deep lung. The purpose of this study was to determine aerosol parameters and post nebulization drug activity of a novel formulation of recombinant Interferon Gamma (rIFNγ), a pluripotent molecule with Th1 helper cell activity. METHODS: rINFγ (200 μg/mL) was aerosolized using i-NEB-MiniTM driven by a portable compressor at 3.5 L/min. Two (2) mL was aerosolized into a Next Generation Impactor (NGI) using a 15 L/min vacuum. Aerosol parameters (MMAD, GSD, Total Respirable Dose) were determined using an HPLC assay (Vectura Ltd., UK). Bioactivity pre-/post-nebulization was determined using an HLADR assay (PBL Assay Sciences, Piscataway, NJ, USA). RESULTS: The MMAD, GSD and Total Respirable dose from a 200 μg/mL formulation (n=6) was 2.5 (± 0.2), 1.6 (± 0.4) and 123.7 (± 19.8) μg respectively. The bioactivity pre- and post-nebulization was consistent with the EP standard suggesting that nebulization did not affect biological activity of the protein. CONCLUSIONS: This study indicates that a clinically relevant and reproducible dose of rINFγ can be delivered with i-NEB-MiniTM. Bioactivity assay indicates that aerosol rINFγ retains protein integrity after nebulization. CLINICAL IMPLICATIONS: rINFγ is an immunomodulatory pleiotropic cytokine with potential for treatment of respiratory diseases such as Pulmonary Fibrosis, MDRTB, COPD and SARS-CoV-2. The results indicate that a clinically relevant dose of rINFγ can be aerosolized using the i-NEB-MiniTM jet nebulizer. DISCLOSURE: Stony Brook and New York University hold patents on the use of inhaled interferon licensed to InspiRx, Inc. Dr. Smaldone consults to InspiRx and is a member of the advisory board. Mr. Shukla and Dr. Toddywala are employees of InspiRx. KEYWORD: immunomodulation
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Introduction/Objective: Sarcoidosis is a syndrome of unknown cause that may manifest with clinical, radiographic and pathological findings similar to those seen with histoplasmosis. We present a case of disseminated histoplasmosis in an immunocompetent patient previously diagnosed with sarcoidosis. Methods/Case Report: A 69-year-old obese male with a history of hypertension, diabetes mellitus and long-standing sarcoidosis was admitted to the hospital for several months of intermittent fevers and pancytopenia. His sarcoidosis was diagnosed 21 years prior, initially involving the lungs and eventually showing cardiac involvement, requiring a pacemaker. He had been treated with methotrexate and prednisone. His recent medical history was also significant for COVID-19 infection, diagnosed 3 months before admission. His fevers were initially attributed to sarcoidosis and his pancytopenia to methotrexate. However, his symptoms continued despite discontinuation of his medications, and further workup was initiated. Computed tomography showed hepatomegaly, splenomegaly, and lymphadenopathy, concerning for a lymphoproliferative disorder. The patient underwent a bone marrow biopsy that showed noncaseating granulomas and microorganisms consistent with histoplasmosis on fungal stain. Bone marrow cultures were not possible as the marrow was inaspirable. The patient subsequently underwent a lymph node biopsy with both morphology and culture identifying histoplasmosis. Urine and serum histoplasma antigen also returned positive. The patient's overall clinical picture was consistent with disseminated histoplasmosis and he was administered intravenous Amphotericin B for 3 weeks followed by oral itraconazole for 1 year. One month follow-up after discharge showed significant improvement in the patient's condition. Results (if a Case Study enter NA): N/A Conclusion: Sarcoidosis reduces T-cell activity, and treatment with steroids causes further immunosuppression and vulnerability for development of a disseminated infection. COVID-19 also presumably increases the predisposition to acquire bacterial or fungal co-infections. Clinicians and pathologists should be aware of the overlap in clinical, radiologic and pathological presentations of sarcoidosis and histoplasmosis to make the correct diagnosis and administer the appropriate treatment.
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Background: The coronavirus-19 disease (COVID-19) pandemic reminds us of the importance of immune function, even in immunologically normal individuals. Multiple lifestyle factors are known to influence the immune function. Objective: The aim was to investigate the association between NK cell activity (NKA) and multiple factors including vitamin D, physical exercise, age, and gender. Methods: This was a cross-sectional association study using health check-up and NKA data of 2,095 subjects collected from 2016 to 2018 in a health check-up center in the Republic of Korea. NKA was measured using the interferon-γ (IFN-γ) stimulation method. The association of NKA with 25-(OH)-vitamin D (25(OH)D) and other factors was investigated by multiple logistic regression analysis. Results: The average age of subjects was 48.8 ± 11.6 years (52.9% of subjects were female). Among 2,095 subjects, 1,427 had normal NKA (NKA ≥ 500 pg IFN-γ/mL), while 506 had low NKA (100 ≤ NKA < 500 pg/mL), and 162 subjects had very low NKA (NKA < 100 pg/mL). Compared to men with low 25(OH)D serum level (< 20 ng/mL), vitamin D replete men (30-39.9 ng/mL) had significantly lower risk of very low NKA (OR: 0.358; 95% CI: 0.138, 0.929; P = 0.035). In women, both low exercise (OR: 0.529; 95% CI: 0.299, 0.939; P = 0.030) and medium to high exercise (OR: 0.522; 95% CI: 0.277, 0.981; P = 0.043) decreased the risk compared to lack of physical exercise. Interestingly, in men and women older than 60 years, physical exercise significantly decreased the risk. Older-age was associated with increased risk of very low NKA in men, but not in women. Conclusion: Physical exercise and vitamin D were associated with NKA in a gender- and age-dependent manner. Age was a major risk factor of very low NKA in men but not in women.