Ein Jahr mit Corona – Stimmen aus der Goethe-Universität

Mitte März des letzten Jahres erreichte die Corona-Pandemie dann auch die Goethe-Universität: Der Lehrbetrieb in den folgenden Semestern wurde und wird hauptsächlich im digitalen Modus durchgeführt, viele Mitarbeiter*innen arbeiten seitdem im Homeoffice, Hygiene- und Abstandsregeln gelten in Räumen und auf Plätzen. In kürzester Zeit mussten Arbeitsabläufe neu organisiert und viele Services auf digitale Prozesse umgestellt werden. Aber das Wichtigste: Die Infektionszahlen konnten gering gehalten werden. Der UniReport hat einige Hochschulangehörige aus Wissenschaft, Verwaltung und Studierendenschaft nach ihren Erfahrungen und Erkenntnissen in diesem außergewöhnlichen Jahr befragt

Induction therapy with Erlotinib (E) and Gemcitabine/Platinum (GP) in stage III NSCLC

Background: In 2004 we started a phase II trial in non-small lung cancer (NSCLC), stage III, with erlotinib followed by a combination with a platinum-based doublet in unselected patients to identify molecular subgroups benefitting from an EGFR targeting approach. Patients and methods: Induction with erlotinib (E, 150 mg, d1-42) was followed by three cycles of gemcitabine (G, 1250 mg/m², d1+d8, q3w) and cisplatin (P, 80 mg/m², d1, q3w). Patients with at least stable disease after E were treated with a GP + E combination. Induction was followed by surgery and radiation. The trial was conducted as a prospective, multi-center, open label, exploratory phase II study to determine pathological response rate (pRR), as well as secondary endpoints disease free survival (DFS) and overall survival (OS). Results: Of 38 prescreened patients 16 were included in the main study. Due to slow recruitment the study had to be terminated early. Combination of E and GP was well tolerated, surgery was feasible after induction therapy in 12 of 16 patients, 7/12 (58%) patients had a major pathological response (MPR). Median overall survival for patients with MPR was 57.7 months (confidence interval (CI), 37.4 to 78.0; n = 7) and for patients without MPR 11.9 months (CI, 6.4 to 17.4; n = 5). 2/16 patients had an epidermal growth factor receptor (EGFR) mutation. Conclusion: Before discovery of distinct molecular mechanisms in NSCLC our study was an attempt to identify clinical and pathological subgroups that would benefit from E induction. Two patients with an EGFR mutation were identified. MPR was a predictor of long term disease free and overall survival

Overnight resting of PBMC changes functional signatures of antigen specific T- cell responses: impact for immune monitoring within clinical trials.

Polyfunctional CD4 or CD8 T cells are proposed to represent a correlate of immune control for persistent viruses as well as for vaccine mediated protection against infection. A well-suited methodology to study complex functional phenotypes of antiviral T cells is the combined staining of intracellular cytokines and phenotypic marker expression using polychromatic flow cytometry. In this study we analyzed the effect of an overnight resting period at 37 °C on the quantity and functionality of HIV-1, EBV, CMV, HBV and HCV specific CD4 and CD8 T-cell responses in a cohort of 21 individuals. We quantified total antigen specific T cells by multimer staining and used 10-color intracellular cytokine staining (ICS) to determine IFNγ, TNFα, IL2 and MIP1β production. After an overnight resting significantly higher numbers of functionally active T cells were detectable by ICS for all tested antigen specificities, whereas the total number of antigen specific T cells determined by multimer staining remained unchanged. Overnight resting shifted the quality of T-cell responses towards polyfunctionality and increased antigen sensitivity of T cells. Our data suggest that the observed effect is mediated by T cells rather than by antigen presenting cells. We conclude that overnight resting of PBMC prior to ex vivo analysis of antiviral T-cell responses represents an efficient method to increase sensitivity of ICS-based methods and has a prominent impact on the functional phenotype of T cells

Overnight resting does not alter the total number of epitope specific T cells.

<p>(A) Frequencies (left panel) and functionality (right panel) of HIV and CMV specific CD8 T cells were determined by ICS. Data indicate numbers of functional T cells as% of total CD8 T cells. Pie charts show the relative contribution of each functional subpopulation within the total CD8 T-cell response according to their functionality. (B) Frequencies of HIV and CMV specific CD8 T cells determined by multimer staining. Data indicate numbers of multimer-positive T cells as% of total CD8 T cells. (A) and (B): data from five HIV (n = 3) or CMV (n = 2) seropositive individuals are shown; median indicated by black line. (C) Comparison of T cell ICS and multimer staining. Bar charts show frequencies of functional epitope specific CD8 T cells determined by ICS calculated as percentage of corresponding multimer-positive T cells. Representative results of two individuals with known epitope specificities (HLA-B8 restricted HIV Nef and HLA-A2 restricted CMV IE1, respectively) are shown. All analyses were performed on not rested and rested PBMC as indicated.</p

Antibodies used for ICS of whole blood.

<p>Antibodies used for ICS of whole blood.</p

Antibodies used for ICS of PBMC.

<p>Antibodies used for ICS of PBMC.</p

Resting effect is not mediated by antigen presenting cells.

<p>(A) Autologous EBV-transformed B-lymphoblastoid cell lines were used as antigen presenting cells to stimulate antigen specific T cells of an EBV seropositive subject. PBMC without the addition of EBV-transformed B-lymphoblastoid cell lines provided the negative controls used for background subtraction. (B) CD3 T cells of a CMV seropositive subject were isolated by magnetic cell sorting. After cryopreservation, CD3 T cells were stimulated directly or after overnight rest with a pool of overlapping peptides corresponding to the CMV-IE-1 protein. (A) and (B) IFNγ, IL2 and MIP1β production was determined by ICS directly or after an overnight resting. Frequencies (bar charts) and functional composition (pie charts) of EBV (A) and CMV (B) specific CD8 T cells are shown. CD8 T-cell subpopulations are depicted according to their functionality (three functions: green; two functions: blue; monofunctional cells: grey). One representative experiment is shown.</p