25-Hydroxyvitamin D concentrations do not affect the humoral or cellular immune response following SARS-CoV-2 mRNA vaccinations.

BACKGROUND: To improve effectiveness of vaccination against SARS-CoV-2, it is important to identify factors that influence the immune response induced by vaccination. Evidence for the role of vitamin D in immune response against SARS-CoV-2 is contradictory. It is therefore of interest whether 25-hydroxyvitamin D (25[OH]D) concentrations affect the humoral and/or cellular response following SARS-CoV-2 vaccination. METHODS: In this prospective cohort study, blood samples were collected from 98 SARS-CoV-2 naive health care workers (HCW) receiving the first two doses of either BNT162b2 or mRNA-1273 in 2021. Wild-type spike (S) protein binding and neutralizing antibodies were determined approximately three weeks after the first dose and four to five weeks after the second dose. Antigen specific T-cells and functionality (proliferative response and interferon gamma [IFN-γ] release) were determined in 18 participants four weeks after the second dose of BNT162b2. We studied the association between 25(OH)D concentrations, which were determined prior to vaccination, and humoral and cellular immune responses following vaccination. RESULTS: We found no association between 25(OH)D concentrations (median 55.9 nmol/L [IQR 40.5-69.8]) and binding or neutralizing antibody titers after complete vaccination (fold change of antibody titers per 10 nmol/L 25(OH)D increase: 0.98 [95% CI 0.93-1.04] and 1.03 [95% CI: 0.96-1.11], respectively), adjusted for age, sex and type of mRNA vaccine. Subsequently, continuous 25(OH)D concentrations were divided into commonly used clinical categories (<25 nmol/L [n = 6, 6%], 25-49 nmol/L [n = 33, 34%], 50-75 nmol/L [n = 37, 38%] and ≥75 nmol/L [n = 22, 22%]), but no association with the humoral immune response following vaccination was found. Also, 25(OH)D concentrations were not associated with the SARS-CoV-2 specific T cell response. CONCLUSION: No association was found between 25(OH)D concentrations and the humoral or cellular immune response following mRNA vaccination against SARS-CoV-2. Based on our findings there is no rationale to advise vitamin D optimization preceding SARS-CoV-2 vaccination in HCW with moderate vitamin D status.

Next-generation bNAbs for HIV-1 cure strategies.

Despite the ability to suppress viral replication using anti-retroviral therapy (ART), HIV-1 remains a global public health problem. Curative strategies for HIV-1 have to target and eradicate latently infected cells across the body, i.e. the viral reservoir. Broadly neutralizing antibodies (bNAbs) targeting the HIV-1 envelope glycoprotein (Env) have the capacity to neutralize virions and bind to infected cells to initiate elimination of these cells. To improve the efficacy of bNAbs in terms of viral suppression and viral reservoir eradication, next generation antibodies (Abs) are being developed that address the current limitations of Ab treatment efficacy; (1) low antigen (Env) density on (reactivated) HIV-1 infected cells, (2) high viral genetic diversity, (3) exhaustion of immune cells and (4) short half-life of Abs. In this review we summarize and discuss preclinical and clinical studies in which anti-HIV-1 Abs demonstrated potent viral control, and describe the development of engineered Abs that could address the limitations described above. Next generation Abs with optimized effector function, avidity, effector cell recruitment and immune cell activation have the potential to contribute to an HIV-1 cure or durable control.

Fine-mapping the immunodominant antibody epitopes on consensus sequence-based HIV-1 envelope trimer vaccine candidates.

The HIV-1 envelope glycoprotein (Env) trimer is the key target for vaccines aimed at inducing neutralizing antibodies (NAbs) against HIV-1. The clinical candidate immunogen ConM SOSIP.v7 is a stabilized native-like HIV-1 Env trimer based on an artificial consensus sequence of all HIV-1 isolates in group M. In preclinical studies ConM SOSIP.v7 trimers induced strong autologous NAb responses in non-human primates (NHPs). To fine-map these responses, we isolated monoclonal antibodies (mAbs) from six cynomolgus macaques that were immunized three times with ConM SOSIP.v7 protein and boosted twice with the closely related ConSOSL.UFO.664 immunogen. A total of 40 ConM and/or ConS-specific mAbs were isolated, of which 18 were retrieved after the three ConM SOSIP.v7 immunizations and 22 after the two immunizations with ConSOSL.UFO.664. 22 mAbs (55%) neutralized the ConM and/or ConS virus. Cross-neutralization of ConS virus by approximately one-third of the mAbs was seen prior to ConSOSL.UFO.664 immunization, albeit with modest potency. Neutralizing antibodies predominantly targeted the V1 and V2 regions of the immunogens, with an apparent extension towards the V3 region. Thus, the V1V2V3 region is immunodominant in the potent NAb response elicited by two consensus sequence native-like HIV-1 Env immunogens. Immunization with these soluble consensus Env proteins also elicited non-neutralizing mAbs targeting the trimer base. These results inform the use and improvement of consensus-based trimer immunogens in combinatorial vaccine strategies.

Antibodies from Rabbits Immunized with HIV-1 Clade B SOSIP Trimers Can Neutralize Multiple Clade B Viruses by Destabilizing the Envelope Glycoprotein.

The high viral diversity of HIV-1 is a formidable hurdle for the development of an HIV-1 vaccine. Elicitation of broadly neutralizing antibodies (bNAbs) would offer a solution, but so far immunization strategies have failed to efficiently elicit bNAbs. To overcome these obstacles, it is important to understand the immune responses elicited by current HIV-1 envelope glycoprotein (Env) immunogens. To gain more insight, we characterized monoclonal antibodies (MAbs) isolated from rabbits immunized with Env SOSIP trimers based on the clade B isolate AMC008. Four rabbits that were immunized three times with AMC008 trimer developed robust autologous and sporadic low-titer heterologous neutralizing responses. Seventeen AMC008 trimer-reactive MAbs were isolated using antigen-specific single B-cell sorting. Four of these MAbs neutralized the autologous AMC008 virus and several other clade B viruses. When visualized by electron microscopy, the complex of the neutralizing MAbs with the AMC008 trimer showed binding to the gp41 subunit with unusual approach angles, and we observed that their neutralization ability depended on their capacity to induce Env trimer dissociation. Thus, AMC008 SOSIP trimer immunization induced clade B-neutralizing MAbs with unusual approach angles with neutralizing effects that involve trimer destabilization. Optimizing these responses might provide an avenue to the induction of trimer-dissociating bNAbs. IMPORTANCE Roughly 32 million people have died as a consequence of HIV-1 infection since the start of the epidemic, and 1.7 million people still get infected with HIV-1 annually. Therefore, a vaccine to prevent HIV-1 infection is urgently needed. Current HIV-1 immunogens are not able to elicit the broad immune responses needed to provide protection against the large variation of HIV-1 strains circulating globally. A better understanding of the humoral immune responses elicited by immunization with state-of-the-art HIV-1 immunogens should facilitate the design of improved HIV-1 vaccine candidates. We identified antibodies with the ability to neutralize multiple HIV-1 viruses by destabilization of the envelope glycoprotein. Their weak but consistent cross-neutralization ability indicates the potential of this epitope to elicit broad responses. The trimer-destabilizing effect of the neutralizing MAbs, combined with detailed characterization of the neutralization epitope, can be used to shape the next generation of HIV-1 immunogens to elicit improved humoral responses after vaccination.

COVA1-18 neutralizing antibody protects against SARS-CoV-2 in three preclinical models.

One year into the Coronavirus Disease 2019 (COVID-19) pandemic caused by Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2), effective treatments are still needed 1-3 . Monoclonal antibodies, given alone or as part of a therapeutic cocktail, have shown promising results in patients, raising the hope that they could play an important role in preventing clinical deterioration in severely ill or in exposed, high risk individuals 4-6 . Here, we evaluated the prophylactic and therapeutic effect of COVA1-18 in vivo , a neutralizing antibody isolated from a convalescent patient 7 and highly potent against the B.1.1.7. isolate 8,9 . In both prophylactic and therapeutic settings, SARS-CoV-2 remained undetectable in the lungs of COVA1-18 treated hACE2 mice. Therapeutic treatment also caused a dramatic reduction in viral loads in the lungs of Syrian hamsters. When administered at 10 mg kg - 1 one day prior to a high dose SARS-CoV-2 challenge in cynomolgus macaques, COVA1-18 had a very strong antiviral activity in the upper respiratory compartments with an estimated reduction in viral infectivity of more than 95%, and prevented lymphopenia and extensive lung lesions. Modelling and experimental findings demonstrate that COVA1-18 has a strong antiviral activity in three different preclinical models and could be a valuable candidate for further clinical evaluation.

Determinants of calcification growth in atherosclerotic carotid arteries; a serial multi-detector CT angiography study.

BACKGROUND: Little is known about the natural course of atherosclerotic plaque in the carotid artery bifurcation. This study investigated the growth pattern of calcifications in atherosclerotic carotid arteries and its determinants using serial multi-detector CT angiography (MDCTA). METHODS: From a cohort of consecutive patients with TIA or ischemic stroke and a baseline MCDTA scan of the carotid arteries, subjects were invited for a follow-up scan after 4-6 years. Calcification volumes were scored semi-automatically on baseline and follow-up scans. Progression of calcification and its determinants were analyzed in two ways: 1. as incidence of newly detectable calcification in patients free of calcification at baseline, using logistic regression analysis; 2. as annual change in calcification volume in all patients, using linear regression analysis. RESULTS: Two-hundred-twenty-two patients (aged 61.0 ± 9.6 years, follow-up time 4.7 ± 0.8 years) were included. Calcification volumes increased significantly (median 2.9 mm³ at baseline versus 9.4 mm³ at follow-up, p < 0.001). Newly detectable calcification during follow-up was found in 27 out of 67 patients without baseline calcification (40.3%) and was independently associated with age (OR 4.6 per 10 years increase in age, p < 0.001) and hypertension (OR 8.2, p = 0.008). Annual calcification growth was independently associated with age, calcification load, glucose, hypertension, and smoking. Baseline calcification load was the most important risk factor for calcification growth in multivariable analysis. CONCLUSION: Several modifiable cardiovascular risk factors are associated with carotid calcification growth, however, time and baseline calcification load remain the most important determinants of calcification development.

Carotid atherosclerotic plaque progression and change in plaque composition over time: a 5-year follow-up study using serial CT angiography.

BACKGROUND AND PURPOSE: Serial in vivo imaging of atherosclerosis is important for understanding plaque progression and is potentially useful in predicting cardiovascular events and monitoring treatment efficacy. This prospective study aims to quantify temporal changes in carotid atherosclerotic plaque volume and plaque composition using MDCTA. MATERIALS AND METHODS: In 109 patients with TIA or ischemic stroke, serial MDCTA of the carotid arteries was performed after 5.3 ± 0.7 years. The carotid bifurcation was semiautomatically registered for paired baseline follow-up datasets. Outer vessel wall and lumen boundaries were defined using semiautomated segmentation tools. Plaque component volumes were measured using HU thresholds. Annual changes in plaque volume and plaque component proportions were calculated. RESULTS: One-hundred-ninety-three carotid arteries were analyzed. Plaque volume decreased in 31% and increased in 69% of vessels (range -5.6-10.1%/year). Overall, plaque volume increased 1.2% per year (95% CI, 0.8-1.6, P ≤ .001). Plaque composition changed significantly from BL (fibrous 66.4%, lipid 28.8%, calcifications 4.8%): fibrous tissue decreased by 1.5%, lipid decreased by 1.8%, and calcification increased by 3.3% (P < .001). Intraobserver reproducibility of all volume and proportion measurements was good (ICC 0.78-1.00) and interobserver reproducibility was moderate (ICC 0.76-0.99). CONCLUSIONS: Changes in carotid plaque burden and plaque composition can be quantified by using serial MDCTA. Plaque burden development is a heterogeneous and slow process.

Evolution of atherosclerotic carotid plaque morphology: do ulcerated plaques heal? A serial multidetector CT angiography study.

BACKGROUND: Atherosclerotic carotid plaque rupture may lead to thromboembolization, causing transient ischemic attack or ischemic stroke. Carotid plaque ulceration on angiography is associated with plaque rupture. Although healing of ruptured plaques has been described in coronary arteries, little is known about the natural development of plaque ulcerations in carotid arteries. We therefore explored the evolution of carotid plaque surface morphology with serial multidetector CT angiography (MDCTA). METHODS: From a registry of patients with transient ischemic attack or minor ischemic stroke, we selected 83 patients who had undergone serial MDCTA of the carotid arteries. Arteries subjected to revascularization procedures between the two scans were excluded (n = 11). Plaque surface morphology was classified as smooth, irregular or ulcerated on both baseline and follow-up MDCTA. Progression (i.e. development of irregularities or ulceration) and regression (i.e. disappearance of irregularities or ulceration) in morphology were evaluated. RESULTS: The mean time interval between the MDCTA scans was 21 ± 13 months. At baseline, 28 (18%) arteries were normal, 124 (80%) contained atherosclerotic plaque and 3 (2%) were occluded. Plaque surface morphology was smooth in 86 arteries (55%), irregular in 23 (15%) and ulcerated in 15 (10%). At follow-up, surface morphology was unchanged in 88% of arteries, had progressed in 8% and regressed in 4%. Most importantly, plaque morphology remained unchanged in most ulcerated plaques (10/15; 67%). One ulcerated plaque had progressed, whereas 4 had regressed. New ulcerations had developed in 2 nonulcerated plaques. CONCLUSION: MDCTA allows evaluation of temporal changes in atherosclerotic carotid plaque morphology. Plaque surface morphology remained unchanged in most arteries. Carotid ulcerations persist for a long time, and may remain a potential source of thromboembolism.

The performance of Bispectral Index in children during equi-MAC halothane vs. sevoflurane anaesthesia.

BACKGROUND AND OBJECTIVE: The reliability of the Bispectral Index for evaluating and monitoring the depth of general anaesthesia in children is not as great as for that in adults. Therefore we analysed Bispectral Index performance in children by comparing changes in Bispectral Index values during a standardized and equipotent anaesthetic regimen using either halothane or sevoflurane for the induction and maintenance of general anaesthesia. Special interest was focussed on excitation during induction, and whether it was associated with simultaneous changes in Bispectral Index scores. METHODS: Twenty children (3-15 yr, ASA I-II) scheduled for general surgery were randomly allocated to either halothane (10 patients) or sevoflurane group (10 patients). Anaesthesia was induced by 3% halothane or 7% sevoflurane, either agent administered with 50% N2O in oxygen for 5 min, the period from the beginning of induction until intubation. Thereafter, anaesthesia was maintained by the respective volatile agent at 1 MAC (minimum alveolar concentration; in addition to 70% N2O in oxygen) and supplemented with remifentanil infusion adjusted to maintain the heart rate and mean arterial pressure to within 20% of the baseline values. Excitation at induction was defined as involuntary muscular movements. RESULTS: Sevoflurane induction produced a more rapid depression in Bispectral Index than halothane, the mean difference being greatest (47 Bispectral Index score) at 105 s. Excitation occurred in three patients during sevoflurane induction, which coincided with increases in Bispectral Index values in two of the three patients. During the maintenance phase at 1 MAC, the Bispectral Index (mean +/- SD) was 57+/-7 for halothane and 47+/-9 for sevoflurane (P < 0.05). The remifentanil doses did not differ between both groups. CONCLUSION: In children, halothane anaesthesia was associated with higher Bispectral Index values than sevoflurane when administered at 1 MAC. Large individual variation in Bispectral Index occurred within both groups. Due to these limitations, one should be cautious when interpreting paediatric Bispectral Index data.

Propofol versus alfentanil to prevent movement responses during uterine curettage.

BACKGROUND: Propofol anaesthesia is frequently associated with movement responses in non-paralysed patients. Opioids decrease the probability of movement during noxious stimulation. Heart rate variability and frontal electromyography (EMG), which are related to subcortical functions, may be more closely related than surface electroencephalography (EEG) to movement responses to noxious stimulation. METHODS: Eighty-two patients scheduled for uterine dilatation and curettage were randomized to receive at the first intra-operative movement either a supplemental alfentanil bolus, 0.5 mg intravenously, or a supplemental propofol bolus, 0.7 mg/kg intravenously. The incidences of recurring movement during the procedure were compared between the two groups. The associations of a measure of heart rate variability (Anemon index), heart rate, EMG and two EEG variables with movement responses were evaluated. RESULTS: The incidences of recurring movement were 73% and 38% in the alfentanil and propofol groups, respectively [difference, 35%; 95% confidence interval, 9-56%; P= 0.014 between the groups). The Anemon index, heart rate, EMG and surface EEG variables displayed mainly reactive associations with movement responses. CONCLUSION: During uterine curettage under propofol-alfentanil-nitrous oxide anaesthesia, a propofol bolus of 0.7 mg/kg was more effective in preventing the recurrence of movement responses than an alfentanil bolus of 0.5 mg. Several physiological variables may be used to track significant arousal reactions, but not to predict them.

EEG spectral entropy, heart rate, photoplethysmography and motor responses to skin incision during sevoflurane anaesthesia.

BACKGROUND: Analgesia is a part of balanced anaesthesia, but direct indicators of nociception do not exist. We examined the relationship between motor reactions and physiological variables during skin incision in sevoflurane anaesthesia and hypothesized that nociception could be detected and graded by significant changes in these variables. METHODS: Thirty-one women scheduled for abdominal hysterectomy participated in the study. Anaesthesia was induced with fentanyl (1 microg kg(-1)), propofol (1 mg kg(-1)) and sevoflurane. Skin incision was performed 14 min after induction during 1.6% end-tidal sevoflurane anaesthesia without neuromuscular blockade. Electrocardiography (ECG), photoplethysmography (PPG) and electroencephalography (EEG) were registered, and a range of variables was computed from these signals. The postincision values, normalized with respect to their preincision values, of movers vs. non-movers were compared. The variables showing significant differences between movers and non-movers were used to develop a logistic regression equation for the classification of patients into movers or non-movers. RESULTS: Twenty-six patients were eligible for analysis, and 12 (46%) displayed a motor reaction to skin incision (movers). Many ECG, PPG and EEG-related variables showed significant differences between the pre- and postincision periods. The best classification performance, assessed by leave-one-out cross-validation, between movers and non-movers was achieved with the combination of response entropy of EEG, RR-interval and PPG notch amplitude. The corresponding equation yielded 96% correct classification with 90% sensitivity and 100% specificity. The classification performance of any single variable alone was considerably worse. CONCLUSION: Combination of information from different sources may be required for monitoring the adequacy of analgesia during anaesthesia.

Identification of peak V in brainstem auditory evoked potentials with neural networks.

A feature extractor for determining the latency of peak V in brainstem auditory evoked potentials (BAEPs) is presented in this paper. A feature extractor that combines artificial neural networks with an algorithmic approach is presented. It consists of a series of small neural networks that have to make simple decisions. Each neural network decides what part of the input pattern contains the peak, and the algorithm passes that part of the pattern to the next neural network; in this way the size of the input patterns decreases during the process, and the last neural network determines the exact location of the peak. An optimal configuration of neural networks could determine the latencies of peak V in all synthetic evoked potentials correctly. With real evoked potentials, the networks yield results that comply with the opinion of a human expert in 80 +/- 6% of the cases.