Class switch toward noninflammatory, spike-specific IgG4 antibodies after repeated SARS-CoV-2 mRNA vaccination.

RNA vaccines are efficient preventive measures to combat the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. High levels of neutralizing SARS-CoV-2 antibodies are an important component of vaccine-induced immunity. Shortly after the initial two mRNA vaccine doses, the immunoglobulin G (IgG) response mainly consists of the proinflammatory subclasses IgG1 and IgG3. Here, we report that several months after the second vaccination, SARS-CoV-2-specific antibodies were increasingly composed of noninflammatory IgG4, which were further boosted by a third mRNA vaccination and/or SARS-CoV-2 variant breakthrough infections. IgG4 antibodies among all spike-specific IgG antibodies rose, on average, from 0.04% shortly after the second vaccination to 19.27% late after the third vaccination. This induction of IgG4 antibodies was not observed after homologous or heterologous SARS-CoV-2 vaccination with adenoviral vectors. Single-cell sequencing and flow cytometry revealed substantial frequencies of IgG4-switched B cells within the spike-binding memory B cell population [median of 14.4%; interquartile range (IQR) of 6.7 to 18.1%] compared with the overall memory B cell repertoire (median of 1.3%; IQR of 0.9 to 2.2%) after three immunizations. This class switch was associated with a reduced capacity of the spike-specific antibodies to mediate antibody-dependent cellular phagocytosis and complement deposition. Because Fc-mediated effector functions are critical for antiviral immunity, these findings may have consequences for the choice and timing of vaccination regimens using mRNA vaccines, including future booster immunizations against SARS-CoV-2.

Dynamics of humoral and cellular immune responses after homologous and heterologous SARS-CoV-2 vaccination with ChAdOx1 nCoV-19 and BNT162b2.

BACKGROUND: Vaccines are an important means to overcome the SARS-CoV-2 pandemic. They induce specific antibody and T-cell responses but it remains open how well vaccine-induced immunity is preserved over time following homologous and heterologous immunization regimens. Here, we compared the dynamics of humoral and cellular immune responses up to 180 days after homologous or heterologous vaccination with either ChAdOx1-nCoV-19 (ChAd) or BNT162b2 (BNT) or both. METHODS: Various tests were used to determine the humoral and cellular immune response. To quantify the antibody levels, we used the surrogate neutralization (sVNT) assay from YHLO, which we augmented with pseudo- and real virus neutralization tests (pVNT and rVNT). Antibody avidity was measured by a modified ELISA. To determine cellular reactivity, we used an IFN-γ Elispot, IFN-γ/IL Flurospot, and intracellular cytokine staining. FINDINGS: Antibody responses significantly waned after vaccination, irrespective of the regimen. The capacity to neutralize SARS-CoV-2 - including variants of concern such as Delta or Omicron - was superior after heterologous compared to homologous BNT vaccination, both of which resulted in longer-lasting humoral immunity than homologous ChAd immunization. All vaccination regimens induced stable, polyfunctional T-cell responses. INTERPRETATION: These findings demonstrate that heterologous vaccination with ChAd and BNT is a potent alternative to induce humoral and cellular immune protection in comparison to the homologous vaccination regimens. FUNDING: The study was funded by the German Centre for Infection Research (DZIF), the European Union's "Horizon 2020 Research and Innovation Programme" under grant agreement No. 101037867 (VACCELERATE), the "Bayerisches Staatsministerium für Wissenschaft und Kunst" for the CoVaKo-2021 and the For-COVID projects and the Helmholtz Association via the collaborative research program "CoViPa". Further support was obtained from the Federal Ministry of Education and Science (BMBF) through the "Netzwerk Universitätsmedizin", project "B-Fast" and "Cov-Immune". KS is supported by the German Federal Ministry of Education and Research (BMBF, 01KI2013) and the Else Kröner-Stiftung (2020_EKEA.127).

Fc-based Duokines: dual-acting costimulatory molecules comprising TNFSF ligands in the single-chain format fused to a heterodimerizing Fc (scDk-Fc).

Targeting costimulatory receptors of the tumor necrosis factor superfamily (TNFSF) to activate T-cells and promote anti-tumor T-cell function have emerged as a promising strategy in cancer immunotherapy. Previous studies have shown that combining two different members of the TNFSF resulted in dual-acting costimulatory molecules with the ability to activate two different receptors either on the same cell or on different cell types. To achieve prolonged plasma half-life and extended drug disposition, we have developed novel dual-acting molecules by fusing single-chain ligands of the TNFSF to heterodimerizing Fc chains (scDuokine-Fc, scDk-Fc). Incorporating costimulatory ligands of the TNF superfamily into a scDk-Fc molecule resulted in enhanced T-cell proliferation translating in an increased anti-tumor activity in combination with a primary T-cell-activating bispecific antibody. Our data show that the scDk-Fc molecules are potent immune-stimulatory molecules that are able to enhance T-cell mediated anti-tumor responses.

A bivalent, bispecific Dab-Fc antibody molecule for dual targeting of HER2 and HER3.

Dual targeting of surface receptors with bispecific antibodies is attracting increasing interest in cancer therapy. Here, we present a novel bivalent and bispecific antagonistic molecule (Dab-Fc) targeting human epidermal growth factors 2 and 3 (HER2 and HER3) derived from the Db-Ig platform, which was developed for the generation of multivalent and multispecific antibody molecules. Dab-Fc comprises the variable domains of the anti-HER2 antibody trastuzumab and the anti-HER3 antibody 3-43 assembled into a diabody-like structure stabilized by CH1 and CL domains and further fused to a human γ1 Fc region. The resulting Dab-Fc 2 × 3 molecule retained unhindered binding to both antigens and was able to bind both antigens sequentially. In cellular experiments, the Dab-Fc 2 × 3 molecule strongly bound to different tumor cell lines expressing HER2 and HER3 and was efficiently internalized. This was associated with potent inhibition of the proliferation and migration of these tumor cell lines. Furthermore, IgG-like pharmacokinetics and anti-tumoral activity were demonstrated in a xenograft tumor model of the gastric cancer cell-line NCI-N87. These results illustrate the suitability of our versatile Db-Ig platform technology for the generation of bivalent bispecific molecules, which has been successfully used here for the dual targeting of HER2 and HER3.

Long-term Effectiveness of Maxillary and Mandibular Bonded Orthodontic Retainers.

PURPOSE: This retrospective cohort study investigated the long-term effectiveness of one type of maxillary and 2 types of mandibular fixed lingual retainers. MATERIALS AND METHODS: Eighty orthodontic patients in retention for 10-15 years were included. Irregularity index, intercanine width, overjet (OJ) and overbite (OB) were measured on plaster models at 3 occasions: (T1) pre-treatment, (T2) post-treatment and (T3) 10-15 years post-treatment. Analyses assessed the effect of the retainer type and time on mandibular irregularity, intercanine width and retainer failure. RESULTS: In the mandible, the irregularity index increased (0.43 mm) between T2 and T3 for the 0.027" ß-titanium (TMA) retainers bonded to canines only while it was stable (-0.02 mm) for the 0.016" x 0.022" braided stainless steel retainers (SS6) bonded to all six anterior teeth. The intercanine width was relatively stable in both groups during the entire observation period. In the maxilla, the irregularity index was stable between T2 and T3 (+0.07 mm). The intercanine width increased (+2.02 mm) during treatment T1-T2 and was stable (-0.02 mm) in the retention phase T2 to T3. CONCLUSIONS: In the mandible, SS6 retainers were slightly more effective in maintaining alignment compared to the TMA retainers. In the maxilla, the SS4 retainers without canine extensions were effective in maintaining alignment. All retainers were effective in maintaining the intercanine width.

Survival of maxillary and mandibular bonded retainers 10 to 15 years after orthodontic treatment: a retrospective observational study.

BACKGROUND: The long-term evidence regarding failures of fixed retainers is limited and the aim of this cohort study was to assess the long-term risk of failure of one type of maxillary and two types of mandibular fixed lingual retainers. TRIAL DESIGN: Retrospective cohort study. METHODS: Eighty-eight patients in retention 10-15 years after orthodontic treatment were included. The type of failure; number of failures per tooth, per patient, and retainer; and adverse effects were assessed by (1) a questionnaire, (2) clinical examination, and (3) screening patients' clinical charts. Descriptive statistics were calculated and a Cox regression was used to assess possible predictors for mandibular retainer survival. RESULTS AND CONCLUSIONS: In the mandible, 47 (53.4%) .016″ × .022″ braided stainless steel retainers (SS) were bonded to all six anterior teeth, and 41 (46.6%) .027″ ß-titanium (TMA) retainers were bonded to the canines only. From the SS retainers 40.4% and of the TMA retainers 61% had no failures during the whole observation period. SS failures per retainer were 2.17 (3.15) vs. 0.66 (1.03) for TMA. The type of retainer was the only significant predictor for failure. In the maxilla, 82 (93.2%) .016″ × .022″ braided SS retainers were bonded to all four incisors and six retainers (6.8%) to all six anterior teeth. The latter group was not further analyzed due to the small sample size. From the retainers bonded to all four incisors, 74.4% had no failure during the whole observation period. SS average number of failures per retainer bonded to the four incisors was 1.14 (SD 2.93). Overall, detachments were the most frequent type of first failure followed by composite damage. From the original mandibular retainers 98.9% and of the original maxillary retainers 97.6% were still in situ 10-15 years after debonding. No adverse torque changes were observed. LIMITATIONS: Potential effects of selection bias, information bias, and attrition bias as well as possible confounding factors cannot be fully excluded in this study.

Judgment of Nasolabial Esthetics in Cleft Lip and Palate Is Not Influenced by Overall Facial Attractiveness.

OBJECTIVE: To determine whether judgment of nasolabial esthetics in cleft lip and palate (CLP) is influenced by overall facial attractiveness. DESIGN: Experimental study. SETTING: University of Bern, Switzerland. SUBJECTS AND METHODS: Seventy-two fused images (36 of boys, 36 of girls) were constructed. Each image comprised (1) the nasolabial region of a treated child with complete unilateral CLP (UCLP) and (2) the external facial features, i.e., the face with masked nasolabial region, of a noncleft child. Photographs of the nasolabial region of six boys and six girls with UCLP representing a wide range of esthetic outcomes, i.e., from very good to very poor appearance, were randomly chosen from a sample of 60 consecutively treated patients in whom nasolabial esthetics had been rated in a previous study. Photographs of external facial features of six boys and six girls without UCLP with various esthetics were randomly selected from patients' files. Eight lay raters evaluated the fused images using a 100-mm visual analogue scale. Method reliability was assessed by reevaluation of fused images after >1 month. A regression model was used to analyze which elements of facial esthetics influenced the perception of nasolabial appearance. RESULTS: Method reliability was good. A regression analysis demonstrated that only the appearance of the nasolabial area affected the esthetic scores of fused images (coefficient = -11.44; P < .001; R(2) = 0.464). The appearance of the external facial features did not influence perceptions of fused images. CONCLUSION: Cropping facial images for assessment of nasolabial appearance in CLP seems unnecessary. Instead, esthetic evaluation can be performed on images of full faces.