Dynamics of immunity over time: decline of anti-SARS-CoV-2 IgG antibodies and T-cell responses after mRNA vaccination in residents and health care workers in nursing homes and homes with assisted living support.

Background: In the present study, we investigated the dynamics of immunity over time by measuring anti SARS-CoV-2 IgG antibodies and SARS-CoV-2 specific T-cell responses (interferon-gamma release assay) after two doses of vaccines in residents and health care workers (HCW). Mostly, 224 (98%) residents and 244 (89%) HCW received two doses of mRNA vaccine (BNT162b2, Pfizer-BioNTech); the rest of the participants received heterologous vaccinations with mRNA and vector vaccines. The study was conducted at the time when the Delta variant of SARS-CoV-2 prevailed. Methods: We analyzed blood samples of 228 residents (median age 83.8 years) and of 273 HCW (median age 49.7 years) from five nursing homes and one home for the elderly with assisted living support at one specific time point. Participants received two vaccinations. The blood samples were analyzed for SARS-CoV-2 specific IgG antibody and T-cell responses. Results: The initial immune responses in the younger participants were about 30% higher than in the older age group. Over time the estimated mean of the parameters (estimated from the study sample for the total population) decreased in all groups within the maximum observation period of 232 days. Comorbidities such as coronary heart disease or diabetes mellitus reduced the initial immune responses regardless of age. With regard to measured IgG antibody levels, absolute values decreased over time, whereas the interferon-gamma response remained at a constant level between day 120 and 180 and seemed to be less dependent on the time elapsed after vaccination. Conclusions: Based on our data, it does not seem possible to determine a reliable threshold of robust immunity, but we suggest that high titres of neutralizing capacity and interferon-gamma response might be an indicator of protection against severe COVID-19 courses.

Long-Term Course of Humoral and Cellular Immune Responses in Outpatients After SARS-CoV-2 Infection.

Characterization of the naturally acquired B and T cell immune responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is important for the development of public health and vaccination strategies to manage the burden of COVID-19 disease. We conducted a prospective, cross-sectional analysis in COVID-19 recovered patients at various time points over a 10-month period in order to investigate how circulating antibody levels and interferon-gamma (IFN-γ) release by peripheral blood cells change over time following natural infection. From March 2020 till January 2021, we enrolled 412 adults mostly with mild or moderate disease course. At each study visit, subjects donated peripheral blood for testing of anti-SARS-CoV-2 IgG antibodies and IFN-γ release after SARS-CoV-2 S-protein stimulation. Anti-SARS-CoV-2 immunoglobulin G (IgG) antibodies were positive in 316 of 412 (76.7%) and borderline in 31 of 412 (7.5%) patients. Our confirmation assay for the presence of neutralizing antibodies was positive in 215 of 412 (52.2%) and borderline in 88 of 412 (21.4%) patients. Likewise, in 274 of 412 (66.5%) positive IFN-γ release and IgG antibodies were detected. With respect to time after infection, both IgG antibody levels and IFN-γ concentrations decreased by about half within 300 days. Statistically, production of IgG and IFN-γ were closely associated, but on an individual basis, we observed patients with high-antibody titres but low IFN-γ levels and vice versa. Our data suggest that immunological reaction is acquired in most individuals after natural infection with SARS-CoV-2 and is sustained in the majority of patients for at least 10 months after infection after a mild or moderate disease course. Since, so far, no robust marker for protection against COVID-19 exists, we recommend utilizing both, IgG and IFN-γ release for an individual assessment of the immunity status.

Antibody Profiling of COVID-19 Patients in an Urban Low-Incidence Region in Northern Germany.

A vast majority of COVID-19 cases present with mild or moderate symptoms. The study region is in an urban and well-defined environment in a low-incidence region in Northern Germany. In the present study, we explored the dynamics of the antibody response with respect to onset, level and duration in patients with confirmed SARS-CoV-2 infection. Anti-SARS-CoV-2 IgG and IgA were detected by automated enzyme-linked immunosorbent assay (ELISA) of SARS-CoV-2 infected patients monitored by the Health Protection Authority. This explorative monocentric study shows IgA and IgG antibody profiles from 118 patients with self-reported mild to moderate, or no COVID-19 related symptoms after laboratory-confirmed infection with SARS-CoV-2. We found that 21.7% and 18.1% of patients were seronegative for IgA or IgG, respectively. Clinically, most of the seronegative patients showed no to only moderate symptoms. With regard to antibody profiling 82% of all patients developed sustainable antibodies (IgG) and 78% (IgA) 3 weeks or later after the infection. Our data indicate that antibody-positivity is a useful indicator of a previous SARS-CoV-2 infection. Negative antibodies do not rule out SARS-CoV-2 infection. Future studies are needed to determine the functionality of the antibodies in terms of neutralization capacity leading to personal protection and prevention ability to transmit the virus as well as to protect after vaccination.

The impact of disturbances in natural conception cycles.

PURPOSE: Many physicians and other healthcare professionals are often asked questions on interfering factors for conception by couples with a desire for children. Such possible disturbances include, for example, the very common minor diseases, stress and also sexual intercourse during the suspected implantation period. Non-scientifically based statements about disturbances in conception cycles, as found in many layman publications and on the internet, can strongly unsettle couples with a desire for children and force them into corset of rules of conduct. Therefore, a systematic scientific evaluation of the impact of disturbances on conception is urgently needed. METHODS: A search for possible disturbances in natural conception cycles together with up to three of the respective pre-cycles in a large cycle database from users of the symptothermal method of natural family planning in Germany was performed. Disturbances were qualified by scientific panel decision and analysed statistically with their effects on the chances of spontaneous conception. Mixed logistical regression models and survival time analyses were used. RESULTS: A total of 237 women with a total of 747 cycles could be included in the analysis. In 61% of all 237 conception cycles, disturbances occurred. The statistical analysis shows that disturbances in natural conception cycles unexpectedly increase the likelihood of pregnancy by an overall factor of 1.32 (95% CI 1.04-1.70). Sexual intercourse in the window of implantation does not decrease the chances of conception. Relaxation states at the time of ovulation and/or during the implantation period have no representable effect and do not increase the chance of pregnancy. CONCLUSIONS: Couples trying to conceive should at least be informed that disturbances in conception cycles, such as minor diseases, stress or sexual intercourse during the implantation period do not interfere with conception. Relaxation has no effect in favour of success. This takes away the guilty feeling of couples, fearing that they possibly did something wrong in cycles without the desired pregnancy.

Luteinising hormone profiles in conception and non-conception natural cycles.

OBJECTIVE: The aim of this study was to assess whether luteinising hormone (LH) surge characteristics influenced the likelihood of conceiving naturally. METHODS: This was a single-cycle, home-based, observational, case-controlled study. Volunteers collected daily urine samples for one menstrual cycle. LH was measured and the basal levels, surge day, peak day, peak concentration and magnitude of LH surges were examined. Predictive models using sociodemographic data, LH surge characteristics, and sociodemographic data combined with LH profile properties, were evaluated. RESULTS: The surge profile did not differ between cycles with early or late ovulation and was not affected by age or body mass index (BMI). The mean LH surge day was day 16 for both groups. Mean LH surge and concentrations did not differ between groups (surge concentration 54.8 IU/l vs. 58.2 IU/l and peak concentration 82.0 IU/l vs. 81.6 IU/l for pregnant vs. non-pregnant volunteers, respectively). Non-pregnant volunteers were more likely, however, to have a raised or a reduced basal LH on day 6 or have an atypical LH profile. Sociodemographic characteristics were significant predictors of pregnancy, and sociodemographic variable-based models had the greatest predictive ability for conception, providing up to 65% predictive accuracy. CONCLUSIONS: Sociodemographic variables can be used to predict the likelihood of a woman conceiving naturally. Provided an LH surge is present, its profile does not relate to the likelihood of spontaneous pregnancy. The conception rate was significantly lower, however, in women with elevated or reduced basal levels of LH, suggesting that follicular maturation needs an optimal basal level of LH in natural conception cycles.

Possible Reasons for Discontinuation of Therapy: an Analysis of 571 071 Treatment Cycles From the German IVF Registry.

Introduction Numerous couples discontinue fertility treatment before achieving the objective, the birth of a child. The aim of this retrospective data analysis is to identify the reasons for early discontinuation of therapy (drop-out). Materials and Methods Retrospective data analysis. With the aid of the German IVF Registry (D·I·R ® ), a total of 122 560 "last cycles" in Germany in the period 2012 - 2015 were identified and the courses were analysed. Results From the named cohort of "last cycles", 37.3% of the female patients (45 699) gave birth to a child and ended the therapy. The remaining 76 861 discontinued the treatment before having a child. The fertility treatment was conducted due to a purely male indication in 46.27% of cases and in 17.96% the cause lay exclusively with the woman. 4.53% of the drop-outs suffered a miscarriage in the last cycle. 73.56% of the drop-out patients ended the therapy after the lack of a positive pregnancy test. After the third therapy cycle, 67% of the couples ended their treatment. Conclusion The results make it possible to provide couples with individual counselling. They offer an option for preparing for the emotional and physical hurdles.

Relationship between anti-Müllerian hormone and antral follicle count across the menstrual cycle using the Beckman Coulter Access assay in comparison with Gen II manual assay.

BACKGROUND: The study aim was to validate Beckman Coulter's fully automated Access Immunoassay System (BC Access assay) for anti-Müllerian hormone (AMH) and compare it with Beckman Coulter's Modified Manual Generation II assay (BC Mod Gen II), with regard to cycle AMH fluctuations and antral follicle counts. METHODS: During one complete menstrual cycle, transvaginal ultrasound was performed on regularly menstruating women (n=39; 18-40years) every 2 days until the dominant ovarian follicle reached 16mm, then daily until observed ovulation; blood samples were collected throughout the cycle. Number and size of antral follicles was determined and AMH levels measured using both assays. RESULTS: AMH levels measured by the BC Access assay vary over ovulatory menstrual cycles, with a statistically significant pre-ovulatory decrease from -5 to +2 days around objective ovulation. Mean luteal AMH levels were significantly lower (-7.99%) than mean follicular levels but increased again towards the end of the luteal phase. Antral follicle count can be estimated from AMH (ng/mL, BC Access assay) concentrations on any follicular phase day. BC Access assay-obtained AMH values are considerably lower compared with the BC Mod Gen II assay (-19% on average); conversion equation: AMH BC Access (ng/mL)=0.85 [AMH BC Mod Gen II (ng/mL)]0.95. CONCLUSIONS: AMH levels vary throughout the cycle, independently of assay utilised. A formula can be used to convert BC Access assay-obtained AMH levels to BC Mod Gen II values. The number of antral follicles can be consistently estimated from pre-ovulatory AMH levels using either assay.

Antimüllerian hormone levels and numbers and sizes of antral follicles in regularly menstruating women of reproductive age referenced to true ovulation day.

OBJECTIVE: To assess menstrual cycle antimüllerian hormone (AMH) levels in reproductive age women and which/how many follicles substantially produce AMH. DESIGN: Prospective study of menstruating women using mixed-effects models to analyze AMH variability and correlation of follicle counts/size classes to AMH levels. SETTING: Clinic. PATIENT(S): Regular menstruating women with ovulatory cycles (n = 40, aged 18-37 years) and no known subfertility. INTERVENTION(S): Women collected daily urine samples and visited the study center for blood samples/transvaginal ultrasound during one complete menstrual cycle (visits were every 2 days; daily from follicle size >16 mm until postovulation). MAIN OUTCOME MEASURE(S): AMH levels throughout the menstrual cycle, correlated with antral follicles as observed by ultrasound and identification of follicles producing AMH. RESULTS: Of all antral follicles visible by high-resolution ultrasound, AMH is produced substantially only by follicles up to 7 mm in diameter. For women with basal AMH >1 ng/mL, mean AMH concentrations vary across ovulatory menstrual cycles, showing a statistically significant decrease from -5 to 2 days after objective ovulation; significantly lower mean luteal AMH levels (-7.59% to mean follicular AMH) are detected. The number of antral follicles can be estimated from AMH (ng/mL) levels using the modified Beckman Coulter Generation II AMH assay for any day of the follicular phase. CONCLUSION(S): AMH concentrations vary across ovulatory menstrual cycles, showing a significant periovulatory decrease. The number of small antral follicles can be estimated from preovulatory AMH levels with relevance for patient management. CLINICAL TRIAL REGISTRATION NUMBER: NCT01802060.

Monitoring the menstrual cycle: Comparison of urinary and serum reproductive hormones referenced to true ovulation.

OBJECTIVE: The aim of the study was to examine relationships and interindividual variations in urinary and serum reproductive hormone levels relative to ultrasound-observed ovulation in menstrual cycles of apparently normally menstruating women. METHODS: This was a prospective study of normally menstruating women (no known subfertility), aged 18-40 years (n = 40), who collected daily urine samples and attended the study centre for blood samples and transvaginal ultrasound during one complete menstrual cycle. Serum luteinising hormone (LH), progesterone, estradiol, urinary LH, pregnanediol-3- glucuronide (P3G) and estrone-3-glucuronide were measured. Ultrasound was conducted by two physicians and interpreted by central expert review. RESULTS: Menstrual cycle length varied from 22 to 37 days (median 27 days). Ovulation by ultrasound ranged from day 8 to day 26 (median day 15). Serum and urinary hormone profiles showed excellent agreement. Estrogen and LH hormone peaks in urine and serum showed a range of signal characteristics across the study group before and after ovulation. The rise in estrogen and LH always occurred before ovulation; the progesterone rise from baseline always occurred after ovulation. CONCLUSIONS: Urinary and serum reproductive hormones showed excellent agreement and may be used interchangeably. The beginning of the surge in serum and urinary LH was an excellent predictor of ovulation. The rise in progesterone and P3G above baseline was a consistent marker of luteinisation confirming ovulation. Both LH and progesterone surges delivered clear, sharp signals in all volunteers, allowing reliable detection and confirmation of ovulation.

The Duesseldorf warning signs for primary immunodeficiency: is it time to change the rules?

OBJECTIVE: Different sets of warning signs can be used if primary immunodeficiency (PID) is suspected: those of the Jeffrey Modell Foundation (JMF), the German Patients' Organisation for Primary Immunodeficiencies (DSAI) and the Association of the Scientific Medical Societies in Germany (AWMF). A few studies have tested the JMF criteria, with unconvincing results, but the diagnostic models of the DSAI and AWMF have not been tested at all. We set out to establish the utility of these three scoring systems and compare them with our own set of five warning signs (Duesseldorf criteria). DESIGN: Prospective study. PATIENTS: Two hundred ten patients admitted to our hospital between 2010 and 2012 with suspected PID. RESULTS: PID were found in 36 (17 %) of the patients admitted. Of the established sets of warning signs, the JMF and the DSAI had inadequate sensitivity, while the DSAI and the AWMF showed insufficient specificity. Our own criteria were analyzed with regard to maximal specificity and sensitivity (Youden Index) and sensitivity and yielded NPV of 0.89 and 0.91 respectively. Youden index revealed combination of five signs and symptoms: lymphopenia, otitis media >7, failure to thrive, failure to grow normally, pneumonia >1. For maximum negative predictive value the following set was found: lymphopenia, hypogammglobulinemia, failure to thrive, growth disorders, iv antibiotics and abscesses. CONCLUSION: In contrast to the new, evaluated Duesseldorf criteria, all three established sets of warning signs proved inadequate for preselection of patients for admission to specialized PID centers. The Duesseldorf criteria should now being tested in further studies.