Immunization of mice with a 14-mer peptide TKDNNLLGRFELSG, termed "TKD," comprising amino acids 450–461 (aa 450–461 ) in the C terminus of inducible Hsp70, resulted in the generation of an IgG1 mouse ...mAb cmHsp70.1. The epitope recognized by cmHsp70.1 mAb, which has been confirmed to be located in the TKD sequence by SPOT analysis, is frequently detectable on the cell surface of human and mouse tumors, but not on isogenic cells and normal tissues, and membrane Hsp70 might thus serve as a tumor-specific target structure. As shown for human tumors, Hsp70 is associated with cholesterol-rich microdomains in the plasma membrane of mouse tumors. Herein, we show that the cmHsp70.1 mAb can selectively induce antibody-dependent cellular cytotoxicity (ADCC) of membrane Hsp70⁺ mouse tumor cells by unstimulated mouse spleen cells. Tumor killing could be further enhanced by activating the effector cells with TKD and IL-2. Three consecutive injections of the cmHsp70.1 mAb into mice bearing CT26 tumors significantly inhibited tumor growth and enhanced the overall survival. These effects were associated with infiltrations of NK cells, macrophages, and granulocytes. The Hsp70 specificity of the ADCC response was confirmed by preventing the antitumor response in tumor-bearing mice by coinjecting the cognate TKD peptide with the cmHsp70.1 mAb, and by blocking the binding of cmHsp70.1 mAb to CT26 tumor cells using either TKD peptide or the C-terminal substrate-binding domain of Hsp70.
Tumor cells, in contrast to normal cells, frequently overexpress heat shock protein 70 (Hsp70) in the cytosol, present it on their cell surface, and actively release it. Therefore, soluble Hsp70 ...(sHsp70) was investigated as a potential tumor biomarker for monitoring the outcome of radiation therapy.
Plasma from mice bearing membrane Hsp70 (mHsp70)-positive FaDu human squamous cell carcinoma of the head and neck and spontaneous pancreatic ductal adenocarcinoma (PDAC) was investigated. A cohort of mice with FaDu tumors (0.32 cm(3)) was irradiated with 30 Gy, and plasma was collected 24 hours after irradiation, after the tumors had shrunk to 50% of their starting volume and after complete remission. sHsp70 levels in the plasma were quantified by enzyme-linked immunosorbent assay.
sHsp70 levels were significantly higher in the blood of tumor-bearing mice than that of control animals. A correlation between increasing sHsp70 plasma levels and tumor volume in the range of 0.01 cm(3) to 0.66 cm(3) was observed. Radiation-induced regression of the tumors was associated with significantly decreased sHsp70 levels, which returned to the level of control animals after complete remission.
We propose sHsp70 as an innovative biomarker for detecting tumors and for monitoring the clinical outcome of radiation therapy in cancer patients.
Membrane-bound heat shock protein 70 (Hsp70) serves as a tumor-specific recognition structure for Hsp70-peptide (TKD) plus IL-2 activated NK cells. A phase I clinical trial has shown that repeated ...re-infusions of ex vivo TKD/IL-2-activated, autologous leukapheresis product is safe. This study investigated the maintenance of the cytolytic activity of NK cells against K562 cells and autologous tumor after 6 plus 3 infusions of TKD/IL-2-activated effector cells.
A stable tumor cell line was generated from the resected anastomotic relapse of a patient with colon carcinoma (pT3, N2, M0, G2). Two months after surgery, the patient received the first monthly i.v. infusion of his ex vivo TKD/IL-2-activated peripheral blood mononuclear cells (PBMNC). After 6 infusions and a pause of 3 months, the patient received another 3 cell infusions. The phenotypic characteristics and activation status of tumor and effector cells were determined immediately before and at times after each infusion.
The NK cell ligands Hsp70, MICA/B, and ULBP-1,2,3 were expressed on the patient's anastomotic relapse. An increased density of activatory NK cell receptors following ex vivo stimulation correlated with an enhanced anti-tumoricidal activity. After 4 re-infusion cycles, the intrinsic cytolytic activity of non-stimulated PBMNC was significantly elevated and this heightened responsiveness persisted for up to 3 months after the last infusion. Another 2 re-stimulations with TKD/IL-2 restored the cytolytic activity after the therapeutic pause.
In a patient with colon carcinoma, repeated infusions of ex vivo TKD/IL-2-activated PBMNC initiate an intrinsic NK cell-mediated cytolytic activity against autologous tumor cells.
Tumor but not normal cells frequently overexpress heat shock protein 70 (Hsp70) and present it on their cell surface (mHsp70) from where it can be actively released. Therefore, membrane (mHsp70) and ...soluble Hsp70 (sHsp70) were investigated as potential tumor biomarkers and for monitoring the outcome of radiation therapy.
Biopsies and blood were collected from patients with squamous cell carcinoma of the head and neck (SCCHN) at different time points (before, during therapy and in the follow-up period). Hsp70 membrane expression was determined on single cell suspensions of tumor biopsies and reference tissues by flow cytometry, sHsp70 protein and antibody levels were determined in the serum of patients and healthy donors by ELISA and NK cell markers that are related to the presence of sHsp70 were analyzed in the patient's peripheral blood lymphocytes (PBL).
Tumor biopsies exhibited significantly increased mHsp70 expression levels compared to the reference tissue. Soluble Hsp70 levels were significantly higher in SCCHN patients compared to healthy human volunteers and high mHsp70 expression levels on tumor cells were associated with high sHsp70 levels in the serum of patients. Following surgery and radiotherapy sHsp70 levels in patients dropped in patients without tumor relapse in the follow-up period. In contrast to sHsp70 protein, anti-Hsp70 antibody levels remained nearly unaltered in the serum of SCCHN patients before and after therapy. Furthermore, sHsp70 protein but not anti-Hsp70 antibody levels were found to be associated with the tumor volume in SCCHN patients before start of therapy. The expression densities of the activatory NK cell markers CD56, CD94, NKG2D, NKp30, Nkp44, and NKp46 differed in patients following therapeutic intervention. A significant increase in the density of NKG2D was observed in SCCHN patients in the follow-up period after surgery and radiotherapy.
We suggest sHsp70 as a potential biomarker for detecting tumors and for monitoring the clinical outcome of radiotherapy in SCCHN patients.
München, Technische Universität München, Diss., 2015
Munich, Technical University of Munich, Diss., 2015
München, Technische Universität München, Diss., 2015
Th17 cells provide protection at barrier tissues but may also contribute to immune pathology. The relevance and induction mechanisms of pathologic Th17 responses in humans are poorly understood. ...Here, we identify the mucocutaneous pathobiont Candida albicans as the major direct inducer of human anti-fungal Th17 cells. Th17 cells directed against other fungi are induced by cross-reactivity to C. albicans. Intestinal inflammation expands total C. albicans and cross-reactive Th17 cells. Strikingly, Th17 cells cross-reactive to the airborne fungus Aspergillus fumigatus are selectively activated and expanded in patients with airway inflammation, especially during acute allergic bronchopulmonary aspergillosis. This indicates a direct link between protective intestinal Th17 responses against C. albicans and lung inflammation caused by airborne fungi. We identify heterologous immunity to a single, ubiquitous member of the microbiota as a central mechanism for systemic induction of human anti-fungal Th17 responses and as a potential risk factor for pulmonary inflammatory diseases.
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•C. albicans is the major fungal inducer of human Th17 responses•Other fungal species rely on cross-reactive Th17 cells against C. albicans•Intestinal inflammation expands C. albicans-specific and cross-reactive Th17 cells•Cross-reactive Th17 cells contribute to A. fumigatus-driven non-intestinal inflammation
Human anti-fungal Th17 immunity and pathology rely on cross-reactivity against Candida albicans.
In recent years, there has been an increasing interest in local infiltration analgesia (LIA) as a technique to control postoperative pain. We compared this technique to the gold standard the 3 in 1 ...femoral nerve block (FNB) in postoperative pain management after total knee arthroplasty (TKA) in a large patient population. This trial analyzes in the early postoperative phase the pain, range of motion, and consumption of pain medications after TKA.
We conducted a retrospective trial that included all patients who were undergoing primary TKA by one single surgeon in a high-volume arthroplasty center in 2015. Patients who have secondary osteoarthritis due to rheumatoid arthritis or previous knee arthrotomy, as well as revision cases, were excluded. The included patients were divided into 2 groups according to the applied pain management (group 1 FNB, group 2 LIA). Concerning the LIA group, a modified form of composition compared to the first describer without the use of adrenaline was carried out. Post-operative additional pain medications were given on a fixed scheme to the patient. The primary outcome was pain at rest over 7 days after surgery labeled by the numeric pain rating scale (NRS). The secondary outcome measures were the total amount of opioid consumption over the hospital stay and the additional need for non-opioid medication. The conversion of the opiate medications on the morphine preparation was carried out according to the conversion data from the literature. For functional recovery, we compared the range of motion in both groups, which was recorded from the second postoperative day by the attending physiotherapist.
In total, 202 patients were assessed for eligibility and included in this clinical trial. Hundred patients were allocated to the continuous FNB group (group 1) and 102 patients to the LIA group (group 2). No statistical difference was found between the two groups regarding demographic data. Primary outcome measurements: The LIA group had a significantly lower NRS score than the continuous FNB group for the measurement in the morning on days 1, 2, and 3 after surgery (day 1, 1.5; day 2, 1.6; day 3, 1.3; p < 0.05). Secondary outcome measurements: The total volume of morphine consumption for the first six postoperative days was significantly lower in the LIA group than the FNB group (FNB 159.8 vs. LIA 96.07). There is also a significant difference between the total morphine consumption of both groups in the direct postoperative course with respect to time and group (two way ANOVA, p < 0.05) On the day of the operation and on the first postoperative day, the intake of additional non-opioids in the LIA group was also significantly reduced compared to the FNB group. No significant difference was observed on the second to sixth postoperative day concerning an additional consumption of non-opioid medications. In terms of range of motion, the LIA group showed a higher active range of motion at the operated extremity than the FNB group during the hospital stay.
The local intraarticular infiltration therapy (LIA) is a sufficient alternative to regional anesthesia avoiding the known risks of regional procedures. The results of this study reflect the efficiency of this pain management with a lower consumption of analgesics, identical to reduced postoperative pain ratings and an improved ROM in the first postoperative days.
Retrospective trial.
To evaluate prevalence and clinical impact of VHL mutations and deletions (3p), a cohort of consecutive kidney tumors was analyzed by DNA sequencing and fluorescence
hybridization (FISH).
The study ...includes 1,805 patients with renal tumors who were surgically treated at the Department of Urology at the University Medical Center Hamburg-Eppendorf between 1994 and 2015. The cohort included 1,176 clear cell, 270 papillary, 101 chromophobe, and 28 clear cell (tubulo) papillary cancers, as well as 149 oncocytomas and 81 less common subtypes.
Among 431 successfully analyzed tumors, VHL mutations were found in 59.3% of clear cell, 5.2% of papillary, 3.1% of chromophobe carcinomas and in 7.3% of oncocytomas as well as in the rare kidney tumor types (25%-60%). FISH analysis was successful in 1,403 cases. 3p25 deletion was found in 57.2% of clear cell, 17.6% of papillary, 17.7% of chromophobe carcinomas and in 11.9% of oncocytomas as well as in the rare kidney tumor types (16.7%-50%). No statistically significant associations between VHL mutation/deletion and tumor grade, stage, and clinical outcome was found. Only in the subgroup of papillary cancers, 3p deletion was significantly associated with lymph node and distant metastasis as well as with poor patient outcome (
< 0.05 each).
The presence of a VHL mutation in virtually all renal tumor subtypes suggests that VHL analysis cannot be used to distinguish between renal tumor subtypes. Consequently, anti-VHL treatment strategies should not be limited to patients with clear cell cancer.
Aims and Objectives:
In recent years, there has been an increasing interest in local infiltration analgesia (LIA) as a technique to control postoperative pain. We compared this technique to the gold ...standard using the 3 in 1 nerve block in postoperative pain management after total knee arthroplasty (TKA). This trial analyzed pain, range of motion and consumption of pain medications after TKA in the early postoperative phase.
Materials and Methods:
We conducted a retrospective trial by analyzing the data of 202 patients, which were separated in two groups. Group 1 treated by 3-in-1 femoral nerve block included 100 patients whereas 102 patients were treated by LIA in group 2. The demographic data, as well as the American Society of Anesthesiologists Score (ASA Score) were collected. The pain intensity was measured objectively with a numeric rating scale (NRS) in the morning and evening. The pain medication was given according to two protocols with a fix opioid dose for the first 3 days only, additional pain medication could be requested by the patient at any time. Not only the dosage but also the number of additional pain medication was observed. The range of motion was measured from the second postoperative day.
Results:
This study showed no statistical difference between the two groups regarding sex, operated side and ASA-score. The pain intensity showed statistical difference between the groups, with the LIA group showing a lower pain intensity in the early postoperative phase, especially in the first days and in the morning. On the 1st postoperative day the average of the numeric pain scale for the nerve block group was 2,3 (SD = 1,6), which was significant higher than the average of the LIA group 1,5 (SD = 1,1). The measurements in the evening show that during the first seven postoperative days there was no significant difference for the pain intensity. The average dose of Oxycodon received on the first postoperative day was 17,8 mg (SD = 9,1) in the nerve block group and 11,5 mg (SD = 6,2) in the LIA group, on the 6th day the dosage received was 10,9 mg (SD = 11,3) respectively 6,0 mg (SD = 7,3). On the first postoperative day 50% of the nerve block group received no additional non-opioid medication, 36% received one additional and 14% received two additional non-opioid medication. In the LIA group 67% received no additional medication, 32% received one additional and only 1% received two additional non-opioid medication. And finally the LIA group showed a statistical better range of motion of the operated knee in the early postoperative phase, on the 2nd day 46° +- 10 versus 36° +- 7 with p = 0,000 and on 7th day 79° +- 14 versus 73° +- 12 with p = 0,016.
Conclusion:
The LIA group showed in this study a significant lower consumption dose of opioid and a quicker recovery of range of motion then the 3 in 1 nerve block group. Furthermore additional pain medication could be reduced during the hospital stay.
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