ABSTRACT
INTRODUCTION: Following SARS-CoV-2-infection up to 21% of patients will develop post-COVID-syndrome. Autoantibodies (AAbs) targeting neuronal-ß-adrenergic and muscarinic receptors may provide crucial contributions to the pathophysiology of this condition. Immunoadsorption (IA) has been identified as an effective means of removing AAbs and has resulted in clinical improvements of other autoantibody-associated diseases. METHODS: We determined AAb-levels (anti-ß1/ß2 and anti-M3/M4 receptor) in 178 patients diagnosed with post-COVID-syndrome and described the clinical courses of two patients with elevated AAb-levels that underwent IA-treatment. RESULTS: AAbs were detected in 57% (101/178) of patients diagnosed with post-COVID-syndrome. Substantial reductions in AAb-levels and clinical remission were achieved in one of two patients who was treated with IA. However, this patient relapsed within 6 weeks with a concomitant increase in AAb-levels. CONCLUSION: Collectively, AAbs may play a pathophysiologic role in post-COVID and their removal provide transient benefits in some patients. However, these findings should be further investigated in randomized-controlled-trials.
Subject(s)
COVID-19 , Humans , Autoantibodies , COVID-19/therapy , SARS-CoV-2 , SyndromeABSTRACT
Background: Treatments are being developed against severe Covid-19 symptoms, among them the use of convalescent plasma. Two drawbacks are, the large volumes of plasma needed for treatment can lead to circulatory overload, and the plasma contains unnecessary components that can lead to unwanted side-effects. We investigated the use of immunoadsorption followed by tangential flow filtration as a method to obtain highly concentrated Covid-19 antibody concentrates free of additional plasma components. Method(s): Five convalescent plasma donors (3 men and 2 women) have participated in this study. Two donors donated twice with 1 year gap between the donations. Immunoadsorption was performed with an affinity column adsorber pair (Miltenyi Biotec, Bergisch-Gladbach, Germany). The resulting eluate contained antibodies dissolved in a glycine buffer with a total volume of 1100- 1500mL. Tangential Flow Filtration System was used to concentrate all eluates 15- 21.3-fold using an Omega 30 kD membrane (Pall Corp, Dreieich, Germany) and to exchange the glycine buffer with 0.9% NaCl. Eluates were then filtered through a sterile filter, before storage at 4degreeC and -80degreeC. Result(s): All donors have tolerated the immunoadsorption very well with no side effects. The final product contained between 5mg and 194mg of CoV-2 antibodies per donation at a median end-volume of 61+/-20ml leading to probably eight times more COVID-19 antibodies than in one plasma unit while preserving or even increasing their neutralization capacity up to ten-fold. Glycine levels were reduced to non-hazardous 12.6+/-20 mumol (see table 1). The product was sterile and remained stable for 0.5 to 1 year in storage. In two cases the concentration of SARS-COV2 antibodies even increased during storage. Conclusion(s): Immunoadsorption followed by tangential flow filtration produces CoV2 antibody concentrates of high concentrations without simultaneous removal of unnecessary plasma components. The procedure can be done within one day, including the donation, without compromising the donor's immune system. Whether these donated antibodies can be used as passive immunization in acutely infected patients remains to be elucidated. (Table Presented).
ABSTRACT
Background Messenger ribonucleic acid (mRNA) is a powerful tool for transferring genetic information. Its advantages include potent but transient gene expression without risk of genomic insertion, tailorable immunogenicity to match therapeutic application, and the potential for efficient, scalable manufacturing.1 The recent success of mRNA-based SARSCoV- 2 vaccines has inspired interest in mRNA as a cancer therapy to deliver immunostimulatory molecules and tumor antigens. However, clinical translation is limited by mRNA instability at physiological conditions and inefficient in vivo delivery.2 A reliable, non-toxic, and stabilizing in vivo delivery system for immunotherapeutic mRNA would help to advance mRNA as a viable cancer therapy. Here, we utilized calcium phosphate mineral-coated microparticles (MCMs) as a delivery system for mRNA-lipid complexes (lipoplexes) to transfect melanoma cells. Methods MCMs were prepared as previously described3 by suspending beta-tricalcium phosphate particles in modified simulated body fluid under rotation for 7 days at 37degreeC, refreshing the media daily. MCMs were then washed in deionized water and freeze dried. Custom-synthesized reporter or therapeutic mRNA constructs were complexed with a lipidic transfecting agent through mixing, then resulting lipoplexes were incubated briefly with MCMs to facilitate electrostatic binding to the porous CaP coating (figure 1a). Loaded MCMs or soluble lipoplexes were added to B16F10 murine melanoma cell culture, and transfection was measured through various assays, including fluorescence microscopy, bioluminescence, and enzymelinked immunosorbent assays. Results Scanning electron microscopy was used to verify platelike, porous coating morphology following MCM fabrication (figure 1b). MCMs enhanced transfection of B16F10 melanoma cells compared to soluble mRNA lipoplex delivery. This was demonstrated with reporter constructs encoding enhanced green fluorescent protein (eGFP, figure 1c) and Gaussia luciferase (G-Luc), as well as with a therapeutic construct encoding interleukin 15 (IL-15), a T cell growth factor. Timelapse imaging also revealed more rapid transfection with MCMs. A close proximity of cells to MCMs was observed as necessary for transfection. Conclusions We demonstrated that MCMs efficiently and locally deliver mRNA lipoplexes to melanoma cells and cause elevated levels of protein expression compared to soluble lipoplex delivery. This enhanced delivery profile makes MCMs a potential drug delivery platform for future in vivo tumor studies and clinical translation. (Figure Presented).
ABSTRACT
We report the case of a Jehovah's Witness adolescent patient with immune-mediated thrombotic thrombocytopenic purpura after SARS-Cov2 infection successfully treated without therapeutic plasma exchange (TPE) using caplacizumab, corticosteroids, rituximab, and extracorporeal immunoadsorption (EIA). Further patients for whom TPE is not an option might benefit from this approach.
ABSTRACT
Introduction: Because of the limited donor pool, transplants are being done across the blood group and even HLA incompatibility barriers. But this comes at the cost of increased immunosuppression and the side effects. Effect of this intensified immunosuppression on the incidence of post transplant infections and the type of infection has not been studied extensively. Methods: We retrospectively analysed the incidence of infection in ABO incompatible transplants (ABOi) and compared it with propensity matched cohort of ABO compatible transplants(ABOc) over the same timeframe i.e. 2011 to April 2019. using hospital eHIS record system. Patients were matched with 1:2 ratio (ABOi: ABOc) for age (<60yr, >60yrs),sex, number of previous transplants, pretransplant infections, history of prior immunosuppression, diabetic status, NODAT, and induction agent used. Desensitization protocol for ABO incompatible transplant includes rituximab with double filtration plasmapheresis, plasmapharesis or immunoadsorption to target anti blood group titre of 8. Patient with high immunological risk (e.g.second transplant, HLA incompatible) receive ATG induction while others receive basiliximab induction. Valganciclovir prophylaxis was given only in patients with ATG induction. Results: [Formula presented] [Formula presented] During the study period 89 patients underwent ABOi transplants which were compared with 178 ABOc transplants. (Table1)Mean follow up duration was 50.45months (SD 26.8) in ABOi group and 49.47months (SD28.7) in ABOc group. 17% patients lost to follow up with their last follow up being more than 2 years before. Incidence of overall infections was similar in both the groups (59% (43/89) Vs 44.3% (79/178);p=0.6). (Table2) Incidence of urinary tract infections(UTI)was significantly more in ABOi group vs ABOc group.(23.5% (21/89) vs 11.79% (21/178);p=0.019). Cytomegalovirus infections (CMV) were significantly more in ABOi group 12.3% (11/89) as compared to ABOc group 5% (9/187) (p=0.04). All the patients with CMV infection were CMV IgG positive pretransplant except 2, one from ABOc group who was CMV IgG negative and another from ABOi group who’s pretransplant CMV serology was unavailable. There was no significant difference in incidence of fungal infection, pneumocystis infection, diarrheal infections (other than CMV),pneumonia (other than CMV, PCP, fungal), Herpes, BKV infection. Incidence of post-transplant tuberculosis (3.3% (3/89) Vs 2.8% (5/178);p=1.0) and SARS COV2 infections (12.3% (11/89) vs 9% (16/178);p=0.39 was similar in both the groups. Patient survival was similar in both the groups i.e.95.5% but death censored graft loss was significantly more in ABOi group 0.9% (8/89) as compared ABOc group 0.3% (5/178) p=0.03. Reason of graft loss in all the patients was immunological and not infection. Infection was cause for death in three ABOi patients and four ABOc patients. Conclusions: Overall incidence of infections in ABOi transplants was similar to Abo compatible transplant. Incidence of UTIs and CMV infections were significantly higher in ABOi group. No conflict of interest
ABSTRACT
For more than two years the whole world is suffering from the COVID-19 pandemic. Before introduction of vaccination strategies the administration of fresh frozen plasma from convalescent donors seemed a promising therapeutic approach, especially if administered during the early phase of disease. The outcomes in multicenter trials on huge cohorts, however, did not meet the expectations. This is one reason why German guidelines do not recommend the use of convalescent plasma. One explanation could be varying and often low concentrations of COVID-19 antibodies in a majority of plasma units at the beginning of plasma applications, which could account for the lack of a convincing clinical efficacy. Therefore, we follow a strategy to selectively collect and concentrate human immunoglobulins using immunoadsorption as the method of antibody donation.
ABSTRACT
Introduction: Limited data exist on the immunogenicity of SARS-CoV-2 vaccination in liver transplant (LT) recipients. Homologous vaccination protocol demonstrated suboptimal responses in this population. Several studies showed that heterologous prime-boost approach yielded a better seroconversion rate in healthy individuals. We, therefore, aimed to explore the immunogenicity and safety of heterologous prime-boost immunization with ChAdOx1 nCoV-19 AstraZeneca (AZ) and BNT162b2 Pfizer-BioNTech (BNT) among LT patients. Method: LT recipients receiving SARS-CoV-2 vaccine at King Chulalongkorn Memorial Hospital between April and November 2021 were consecutively enrolled. Patients received either heterologous prime-boost protocol (AZ/BNT) or homologous regimen (AZ/AZ) depending on national policies. Blood samples were collected before vaccine administration and 4 weeks after the second dose. SARS-CoV-2 spike receptor-binding-domain (RBD) IgG was measured using electrochemiluminescence immunoassay (Roche). A titer $100 U/mL was considered as a protective antibody. The neutralizing antibody was detected by enzymelinked immunosorbent assay-based surrogate virus neutralization test (sVNT) (GenScript) with positive cut-off of ≥30% inhibition. Adverse events (AEs) within 7 days following vaccination were recorded by questionnaires. Results: Eighty-nine LT recipients were enrolled. Of these, 64 (71.9%) and 25 (28.1%) patients received AZ/BNT and AZ/AZ, respectively. Majority was male (68.5%) with mean age of 58.3±14.5 years. Median time since transplant was 5.7 years (IQR 2.8-11.8). No patient had a previous diagnosis of SARSCoV- 2 infection. Comparable seroconversion rate of anti-RBD antibody and sVNT after vaccination were observed in both AZ/BNT and AZ/AZ groups (70.3% vs 64.0%, p=0.62 and 77.8% vs 59.1%, p=0.10, respectively). However, median IgG titer was significantly higher in AZ/BNT group compared to AZ/AZ group (842.9 U/mL vs 152.2 U/mL, p=0.011). Patients receiving AZ/BNT also had significantly higher sVNT levels than those receiving AZ/BNT (91.2% vs 35.8%, p=0.003) (Figure 1). In multivariate analysis, use of prednisolone (OR 6.3, 95%CI 1.1-35.4, p=0.036) and LT duration <5 years (OR 3.2, 95%CI 1.1-9.1, p= 0.029) were associated with lack of protective serological response. AZ/BNT regimen trended to have higher reported AEs than AZ/AZ (Figure 2). No graft rejection or serious AEs were found. Conclusion: Heterologous prime-boost regimen with adenoviral vectored and mRNA SARS-CoV-2 vaccine yielded greater humoral response than homologous protocol and was well-tolerated in LT recipients. T-cell response, which might play additional role in protection, and immune durability of this mix-and-match strategy are eagerly awaited. (Figure Presented)
ABSTRACT
BACKGROUND AND AIMS: Recent studies evaluated safety and efficacy of vaccines against coronavirus disease (COVID-19), but none of them are currently approved in Russia. Here, we report results on immunogenicity of the recombinant adenovirus (rAd) 26 and rAd5 vector-based COVID-19 vaccine Gam-COVID-Vac (Sputnik V, developed by Gamaleya National Research Centre, Russia) in patients, receiving maintenance haemodialysis (HD). We aimed to compare the strength of humoral and cellular immunity after 2 doses of Gam-COVID-Vac in patients receiving HD and individuals with normal kidney function. METHOD: The prospective cohort study (NCT: 04 805 632) included 23 patients treated with maintenance HD and 28 volunteers with normal kidney function. All participates were adult, had been fully vaccinated with Gam-COVID-Vac vaccine and had no prior history of suspected or confirmed COVID-19. Subjects were excluded if they had a history of confirmed SARS-CoV-2 infection, had underlying autoimmune disease, malignancies or concomitant immunosuppressive therapy. In all participants the levels of specific IgG were quantified at 4 weeks after second vaccine dose administration using a semi-quantitative SARS-CoV-2 S1 IgG enzymelinked immunosorbent assay. In all subjects, specific CD4+ and CD8+ T-lymphocyte responses (count of spots to spike structural peptide of SARS-CoV-2 virus in IGRA test) were evaluated at the same timepoint. All the participants were asked to report adverse events (AEs) following first and second vaccine administration, included general malaise, fever, myalgia, headache, allergic reactions and injection site reactions. RESULTS: Overall, the incidence of vaccine-associated AEs was less in HD patients than in healthy controls: 32% versus 75% after both first and second vaccine administrations, RR = 0.46 (95% confidence interval 0.24-0.79), P = 009 (Table 1). The most commonly reported AE in both groups was pain in the injection site. No severe or serious AEs occurred in both patients and healthy controls. The seroconversion rate in 4 weeks after second vaccine shot reached 100% (28 of 28) in healthy subjects and 87% (20 of 23) in patients receiving HD. IgG levels did not differ between groups: 4.7 [Q1-Q3: 3.37;6.25] in HD patients versus 5.5 [Q1-Q3: 3.3;7.2] in controls, P = 0.219 (Figure 1A). The T-test result was positive in 70% (16 of 23) of HD patients and in 79% (22 of 28) of controls. The magnitude of T-cell response was comparable between groups: 30 spots [Q1-Q3: 11;48] in HD patients versus 30 spots [Q1-Q3: 14;51] in controls, P = 0.745 (Figure 1B). CONCLUSION: Patients receiving HD develop efficient humoral and cellular immune responses after complete vaccination against COVID-19 with Gam-COVID-Vac vaccine, which is comparable with those in healthy adults.
ABSTRACT
For more than a year the whole world is suffering from the COVID-19 pandemic with no treatment option in sight. Administration of plasma from convalescent donors containing anti-SARS-CoV-2 antibodies, though promising according to case reports, failed to show a clear benefit in a greater number of trials. One reason could be varying and low antibody contents in a majority of plasma units hampering standardization and clinical efficacy. Besides, other plasma components unnecessarily transfused like coagulation factors might promote hypercoagulation seen in severe COVID-19 etiopathology. We therefore hypothesized that instead of collecting whole plasma units, convalescent donors could donate solely immunoglobulins by undergoing immunoadsorption, a mode of therapy regularly applied in autoimmune diseases. Here, we report the results of the first two antibody donations performed at the University Hospital Düsseldorf. In both cases, immunoadsorptions were very well tolerated with no side effects. Collected and neutralized eluates were concentrated using tangential flow filtration increasing the concentration of immunoglobulins 10fold as compared to peripheral blood and leading to probably eight times more neutralizing antibodies than in one plasma unit. Therefore, immunoadsorption can be used as a method of antibody donation. Whether these donated antibodies can be used as passive immunization in acutely infected patients remains to be elucidated.