Herstellung und Charakterisierung monoklonaler Antikörper gegen humanes Interleukin-3

Interleukin-3 ist ein hämatopoetisch und immunmodulatorisch wirkendes Zytokin. In der Kollagen-induzierten Arthritis (CIA), dem Mausmodell der Rheumatoiden Arthritis, hatte die Blockade von Interleukin-3 einen entzündungshemmenden Effekt auf die Frühphase der Erkrankung. Im Rahmen dieser Dissertation wurden neun monoklonale Antikörper gegen humanes Interleukin-3 durch Fusion muriner Milzzellen mit der Myelomzelllinie X63Ag8 hergestellt und diese sowie fünf weitere in der Arbeitsgruppe hergestellte Antikörper charakterisiert. Idealerweise sollten die entstandenen Antikörper den Isotyp IgG aufweisen, gegen ein lineares Epitop gerichtet sein und die biologischen Eigenschaften des IL-3 blockieren. Alle eigens hergestellten Antikörper wiesen den Isotyp IgG auf. Sieben Antikörper waren gegen dreidimensionale Epitope gerichtet, sieben Antikörper gegen lineare Epitope. Die Antikörper binden humanes IL-3 sehr spezifisch: Keiner der Antikörper zeigte eine Kreuzreaktivität gegenüber Ratten-, Maus- oder Rhesusaffen- IL-3. Nur zwei der Antikörper zeigten eine minimale Kreuzreaktivität gegenüber dem strukturverwandten IL-5, einer eine geringe gegenüber GM-CSF. Die Affinitäten von zwei Antikörpern befanden sich im picomolaren, von neun im nanomolaren sowie die zweier weiterer im mikromolaren Bereich. Die blockierende Wirkung der Antikörper wurde anhand eines Proliferationsassays einer IL-3-abhängigen Zelllinie untersucht. Es stellte sich heraus, dass einer der getesteten Antikörper die Proliferation dieser Zellen unterbindet. Weiterhin wurde durchflusszytometrisch untersucht, ob die Antikörper eine Bindung des IL-3 an mononukleäre Zellen des Blutes verhindern. Zwei Antikörper zeigten eine deutliche Blockade der IL-3-Bindung an diese Zellen. Zum Abschluss wurde untersucht, ob die Antikörper die Aktivierung und Zytokfreisetzung durch Basophile Granulozyten unterbinden, da diese Zellreihe für die Wirkung des IL-3 in der CIA verantwortlich gemacht wird. Einer der getesteten Antikörper blockierte die Aktivierung der Basophilen und damit auch die Zytkoinausschüttung beinahe vollständig. Somit zeigte sich, dass einer der getesteten IgG-Antikörper gegen die Sequenz SWVN gerichtet ist und die biologische Wirkung des IL-3 effektiv unterbindet. Dies könnte sowohl für die Rheumatoide Athritis als auch für weitere mit IL-3-assozierte Erkrankungen von Bedeutung sein

The prognostic significance of a negative PSMA-PET scan prior to salvage radiotherapy following radical prostatectomy.

AIM The optimal management for early recurrent prostate cancer following radical prostatectomy (RP) in patients with negative prostate-specific membrane antigen positron-emission tomography (PSMA-PET) scan is an ongoing subject of debate. The aim of this study was to evaluate the outcome of salvage radiotherapy (SRT) in patients with biochemical recurrence with negative PSMA PET finding. METHODS This retrospective, multicenter (11 centers, 5 countries) analysis included patients who underwent SRT following biochemical recurrence (BR) of PC after RP without evidence of disease on PSMA-PET staging. Biochemical recurrence-free survival (bRFS), metastatic-free survival (MFS) and overall survival (OS) were assessed using Kaplan-Meier method. Multivariable Cox proportional hazards regression assessed predefined predictors of survival outcomes. RESULTS Three hundred patients were included, 253 (84.3%) received SRT to the prostate bed only, 46 (15.3%) additional elective pelvic nodal irradiation, respectively. Only 41 patients (13.7%) received concomitant androgen deprivation therapy (ADT). Median follow-up after SRT was 33 months (IQR: 20-46 months). Three-year bRFS, MFS, and OS following SRT were 73.9%, 87.8%, and 99.1%, respectively. Three-year bRFS was 77.5% and 48.3% for patients with PSA levels before PSMA-PET ≤ 0.5 ng/ml and > 0.5 ng/ml, respectively. Using univariate analysis, the International Society of Urological Pathology (ISUP) grade > 2 (p = 0.006), metastatic pelvic lymph nodes at surgery (p = 0.032), seminal vesicle involvement (p 0.5 ng/ml (p = 0.004), and lack of concomitant ADT (p = 0.023) were significantly associated with worse bRFS. On multivariate Cox proportional hazards, seminal vesicle infiltration (p = 0.007), ISUP score >2 (p = 0.048), and pre SRT PSA level > 0.5 ng/ml (p = 0.013) remained significantly associated with worse bRFS. CONCLUSION Favorable bRFS after SRT in patients with BR and negative PSMA-PET following RP was achieved. These data support the usage of early SRT for patients with negative PSMA-PET findings

Development and Validation of a Multi-institutional Nomogram of Outcomes for PSMA-PET-Based Salvage Radiotherapy for Recurrent Prostate Cancer.

IMPORTANCE Prostate-specific antigen membrane positron-emission tomography (PSMA-PET) is increasingly used to guide salvage radiotherapy (sRT) after radical prostatectomy for patients with recurrent or persistent prostate cancer. OBJECTIVE To develop and validate a nomogram for prediction of freedom from biochemical failure (FFBF) after PSMA-PET-based sRT. DESIGN, SETTING, AND PARTICIPANTS This retrospective cohort study included 1029 patients with prostate cancer treated between July 1, 2013, and June 30, 2020, at 11 centers from 5 countries. The initial database consisted of 1221 patients. All patients had a PSMA-PET scan prior to sRT. Data were analyzed in November 2022. EXPOSURES Patients with a detectable post-radical prostatectomy prostate-specific antigen (PSA) level treated with sRT to the prostatic fossa with or without additional sRT to pelvic lymphatics or concurrent androgen deprivation therapy (ADT) were eligible. MAIN OUTCOMES AND MEASURES The FFBF rate was estimated, and a predictive nomogram was generated and validated. Biochemical relapse was defined as a PSA nadir of 0.2 ng/mL after sRT. RESULTS In the nomogram creation and validation process, 1029 patients (median age at sRT, 70 years [IQR, 64-74 years]) were included and further divided into a training set (n = 708), internal validation set (n = 271), and external outlier validation set (n = 50). The median follow-up was 32 months (IQR, 21-45 months). Based on the PSMA-PET scan prior to sRT, 437 patients (42.5%) had local recurrences and 313 patients (30.4%) had nodal recurrences. Pelvic lymphatics were electively irradiated for 395 patients (38.4%). All patients received sRT to the prostatic fossa: 103 (10.0%) received a dose of less than 66 Gy, 551 (53.5%) received a dose of 66 to 70 Gy, and 375 (36.5%) received a dose of more than 70 Gy. Androgen deprivation therapy was given to 325 (31.6%) patients. On multivariable Cox proportional hazards regression analysis, pre-sRT PSA level (hazard ratio [HR], 1.80 [95% CI, 1.41-2.31]), International Society of Urological Pathology grade in surgery specimen (grade 5 vs 1+2: HR, 2.39 [95% CI, 1.63-3.50], pT stage (pT3b+pT4 vs pT2: HR, 1.91 [95% CI, 1.39-2.67]), surgical margins (R0 vs R1+R2+Rx: HR, 0.60 [95% CI, 0.48-0.78]), ADT use (HR, 0.49 [95% CI, 0.37-0.65]), sRT dose (>70 vs ≤66 Gy: HR, 0.44 [95% CI, 0.29-0.67]), and nodal recurrence detected on PSMA-PET scans (HR, 1.42 [95% CI, 1.09-1.85]) were associated with FFBF. The mean (SD) nomogram concordance index for FFBF was 0.72 (0.06) for the internal validation cohort and 0.67 (0.11) in the external outlier validation cohort. CONCLUSIONS AND RELEVANCE This cohort study of patients with prostate cancer presents an internally and externally validated nomogram that estimated individual patient outcomes after PSMA-PET-guided sRT

Minimally Invasive Breast Fibroadenoma Excision Using an Ultrasound-Guided Vacuum-Assisted Biopsy Device

Introduction We studied the extent to which an ultrasound-guided vacuum-assisted biopsy device ("hand-held Mammotome") could be used not only as a valuable tool for investigation of suspicious breast lesions, but also therapeutically for the complete removal of breast fibroadenomas. Materials and Methods 132 aspiration biopsies showing fibroadenoma on histology were collected retrospectively. We ascertained whether there were residual findings on breast ultrasound at a median follow-up of approx. 9 months following biopsy. A questionnaire was used to determine the complication rate, patient satisfaction and acceptance of the procedure. Results In this study complete fibroadenomectomy was achieved at aspiration biopsy in 76% of cases (n = 132). Compared to fibroadenomas larger than 2.1 cm(3) (59%), those smaller than 2.5 cm(3) were completely removed more often (87.6%; p < 0.05). The procedure is associated with very little pain during and after biopsy and minimal haematoma development, both factors supporting a high rate of acceptance among patients. Conclusion Ultrasound-guided vacuum-assisted biopsy is safe and associated with very few complications. Its additional therapeutic potential is dependant on the size of the benign lesion as measured at initial ultrasound

Does post-operative radiochemotherapy improve survival in high-grade endometrial cancer patients? Results of a population-based cohort analysis of a cancer registry

Adjuvant treatment of high-grade endometrial cancer varies greatly due to the lack of definitive results from controlled randomized trials on the subject. In a retrospective study, we sought to investigate the influence of post-operative radio-, chemo, and radiochemotherapy on survival time and recurrence rates among high-grade endometrial cancer patients. 284 high-grade endometrial cancer patients (FIGOI-III, or unknown classification) diagnosed between 1998 and 2015 were retrospectively analyzed. All patients underwent surgery. Overall survival (OS), recurrence-free survival (RFS), and recurrence rates were compared for post-operative treatment modalities of radiotherapy alone (RT), chemotherapy alone (CTX), radiochemotherapy (RCT), and observation (OBS). Post-operative RCT and RT resulted in a significantly improved 5-year OS of 94.1% (HR 0.104, CI 0.013-0.809) and 62.1% (HR 0.615, CI 0.390-0.969), respectively, compared to 43.6% for OBS. CTX did not significantly improve OS leading to a 5-year OS of 56.5% (HR 0.783, CI 0.224-2.740). 5-year recurrence rate was lowest for patients treated with RCT (5.3%). 5-year RFS was 94.1% for the RCT group and proved to be significantly superior to 58.8% for RT (HR 9.034, CI 1.184-68.948), 56% for CTX (HR 12.738, CI 1.337-121.346), and 37.4% for OBS (HR 16.407, CI 2.127-126.575), respectively. In comparison with OBS, RT alone resulted in a significant improvement in RFS (HR 0.551, CI 0.354-0.856). Our retrospective population-based study indicates a survival benefit from treating high-grade endometrial cancer with post-operative RCT. Randomized controlled trials are needed to minimize potential confounding parameters and further clarify the subject

Quality of life and oncological outcome in endometrial cancer patients after vaginal brachytherapy: comparison of two dosing schemes

PurposeVaginal brachytherapy reduces the risk of local recurrence and was shown to be equieffective in preventing vaginal vault recurrence, but less toxic compared to external-beam radiotherapy in a subset of high intermediate-risk endometrial cancer patients and is administered as single adjuvant treatment in those patients. Different radiotherapeutic approaches with various dosing schemes exist toward brachytherapy. The aim of this study was to compare the outcome and long-term quality of life after brachytherapy with two different high-dose-rate dosing schemes.MethodsRetrospective analysis was conducted of the recurrence and survival rates of 104 patients with endometrial cancer FIGO stage I-II that underwent adjuvant brachytherapy with three times 5Gy or four times 5Gy to the upper two-thirds of the vaginal vault in two different institutions between January 2010 and December 2013. Quality of life was assessed by EORTC QLQ-30 questionnaire and EN 24 module.ResultsThe vaginal vault recurrence rates were 4.9% and 5.0% for patients treated with 3 x 5Gy and 4 x 5Gy, respectively (p=0.98). We did not observe a difference in pelvic recurrence (p=0.96), overall survival (p=0.33) or quality of life between the different radiotherapy regimens. Metastatic recurrence and the use of chemotherapy contribute to impairment on quality of life. Younger patients (<70years) reported worse emotional functioning (p=0.02) and higher symptom scales of diarrhea (p=0.01) and financial problems (p=0.03). Sexual activity was lower in patients younger than 70years (p=0.05).ConclusionsFurther prospective studies are needed to evaluate the effect of dosing schemes on recurrence rates and quality of life. Younger patients (<70years) seem to experience greater reduction in quality of life due to endometrial cancer diagnosis

Sentinel Lymph Node Biopsy in Breast Cancer Patients by Means of Indocyanine Green Using the Karl Storz VITOM® Fluorescence Camera

Currently, the use of radioisotope and blue dye for sentinel lymph node biopsy (SLNB) for axillary staging in breast cancer is common. Recently, indocyanine green (ICG) has been proposed as an alternative sentinel lymph node (SLN) tracking agent. We evaluated the clinical value of ICG as an additional tracer in combination with Technetium99m and as an alternative to Technetium99m for the identification of SLN in 104 breast cancer patients. 21 patients had at least 1 histologically tumor-positive SLN. All 21 patients were detected by ICG; in one of these 21 sentinel-positive patients, Technetium99m was unable to identify lymph node involvement. Our results show that ICG is as effective as the radioisotope for SLNB. In addition, as a near-infrared dye, it has the advantages of real-time visualization, lower cost, and wider availability, since no radioactive material needs to be handled. This trial is registered with German Clinical Trial Register Main ID: DRKS00013606

Is there a benefit of lymphadenectomy for overall and recurrence-free survival in type I FIGO IB G1-2 endometrial carcinoma? A retrospective population-based cohort analysis

The recommended therapy for type I FIGO IB endometrial cancer (EC) is hysterectomy and adnexectomy, but the therapeutic benefits of additional pelvic and paraaortic lymph node dissection (LND) are still under discussion. In this study, we retrospectively evaluated overall survival (OAS) and recurrence-free survival (RFS) among patients with type I FIGO IB EC who did undergo systematic or elective lymphadenectomy or none at all. We selected 299 individuals from the database of the German Tumor Centre Regensburg who were diagnosed between 1998 and 2015 with endometrial adenocarcinoma of the uterus type I FIGO IB. We applied multivariable Cox regression to the selected patient data and estimated hazard ratios for OAS and RFS against the performed intervention. Further, we carried out risk adjustments with respect to clinicopathological parameters, and performed model selection using conditional stepwise forward selection. We observed significant benefits of LND in the unadjusted survival analysis; however, we did not confirm this effect in multivariable regression analysis upon risk adjustment. In this case, hazard ratio (HR) for OAS in patients without LND versus patients with LND is reduced to 1.214 (95% CI 0.771-1.911; p = 0.402), HR for RFS is 1.059 (95% CI 0.689-1.626; p = 0.795). Similarly, we were also able to eliminate the statistical benefit of systematic versus elective LND by risk adjustment. In contrast to previous observations in high-grade EC, our study provides compelling evidence that LND, in particular systematic lymphadenectomy, is not beneficial for patients with type I FIGO IB EC in terms of long-term OAS and RFS

Value of indocyanine green pelvic lymph node mapping in the surgical approach of cervical cancer

Purpose Lymph node metastasis is a significant predictive factor for disease recurrence and survival in cervical cancer patients and relevant for therapeutic strategies. We evaluated the clinical value of indocyanine green (ICG) by measuring the sensitivity and negative predictive value of sentinel lymph node mapping compared with the gold standard of complete lymphadenectomy in detecting lymph node metastases for cervical cancer. Methods We utilized the near-infrared imaging agent ICG to detect tumor-infested lymph nodes in the pelvis analogue to a classical sentinel lymph node procedure by analyzing data from 20 patients who had undergone surgery for cervical cancer at our institution. A laparoscopic lymph node mapping procedure by means of ICG, followed by a complete pelvic lymphadenectomy with or without paraaortic lymphadenectomy was done in all patients. Results Histological examination identified seven patients with tumor-positive pelvic nodes, whereas mapping with ICG identified only five of these patients. Detection rate of positive nodes by ICG mapping and false negative rate was 71.4% and 28.6%, respectively; bilateral detection rate was 83.3%. One of the two false negative patients additionally suffered from deep infiltrating endometriosis. Conclusions Our results indicate that ICG can identify the relevant pelvic nodes independent of tumor size, provided bilateral detection is achieved and additional, related diseases are excluded