Tumor-Associated Disialylated Glycosphingolipid Antigen-Revealing Antibodies Found in Melanoma Patients' Immunoglobulin Repertoire Suggest a Two-Direction Regulation Mechanism Between Immune B Cells and the Tumor.

There is far less information available about the tumor infiltrating B (TIL-B) cells, than about the tumor infiltrating T cells. We focused on discovering the features and potential role of B lymphocytes in solid tumors. Our project aimed to develop innovative strategies to define cancer membrane structures. We chose two solid tumor types, with variable to considerable B cell infiltration. The strategy we set up with invasive breast carcinoma, showing medullary features, has been introduced and standardized in metastatic melanoma. After detecting B lymphocytes by immunohistochemistry, VH-JH, Vκ-Jκ immunoglobulin rearranged V region genes were amplified by RT-PCR, from TIL-B cDNA. Immunoglobulin variable-region genes of interest were cloned, sequenced, and subjected to a comparative DNA analysis. Single-chain variable (scFv) antibody construction was performed in selected cases to generate a scFv library and to test tumor binding capacity. DNA sequence analysis revealed an overrepresented VH3-1 cluster, represented both in the breast cancer and the melanoma TIL-B immunoglobulin repertoire. We observed that our previously defined anti GD3 ganglioside-binder antibody-variable region genes were present in melanoma as well. Our antibody fragments showed binding potential to disialylated glycosphingolipids (GD3 ganglioside) and their O acetylated forms on melanoma cancer cells. We conclude that our results have a considerable tumor immunological impact, as they reveal the power of TIL-B cells to recognize strong tumor-associated glycosphingolipid structures on melanomas and other solid tumors. As tumor-derived gangliosides affect immune cell functions and reduce the B lymphocytes' antibody production, we suspect an important B lymphocyte and cancer cell crosstalk mechanism. We not only described the isolation and specificity testing of the tumor infiltrating B cells, but also showed the TIL-B cells' highly tumor-associated GD3 ganglioside-revealing potential in melanomas. The present data help to identify new cancer-associated biomarkers that may serve for novel cancer diagnostics. The two-direction regulation mechanism between immune B cells and the tumor could eventually be developed into an innovative cancer treatment strategy.

Challenging tumour immunological techniques that help to track cancer stem cells in malignant melanomas and other solid tumours.

AIM OF THE STUDY: The arsenal of questions and answers about the minor cancer initiating cancer stem cell (CSC) population put responsible for cancer invasiveness and metastases, has left with an unsolved puzzle. Specific aims of a complex project were partly focused on revealing new biomarkers of cancer. We designed and set up novel techniques to facilitate the detection of cancerous cells. MATERIALS AND METHODS: As a novel approach, we investigated B cells infiltrating breast carcinomas and melanomas (TIL-B) in terms of their tumour antigen binding potential. By developing the TIL-B phage display technology we provide here a new technology for the specific detection of highly tumour-associated antigens. Single chain Fv (scFv) antibody fragment phage ELISA, immunofluorescence (IF) FACS analysis, chamber slide technique with IF confocal laser microscopy and immunohistochemistry (IHC) in paraffin-embedded tissue sections were set up and standardized. RESULTS: We showed strong tumour-associated disialylated glycosphingolipid expression levels on various cancer cells using scFv antibody fragments, generated previously by uniquely invasive breast carcinoma TIL-B phage display library technology. CONCLUSIONS: We report herein a novel strategy to obtain antibody fragments of human origin that recognise tumour-associated ganglioside antigens. Our investigations have the power to detect privileged molecules in cancer progression, invasiveness, and metastases. The technical achievements of this study are being harnessed for early diagnostics and effective cancer therapeutics.

[Genetic information from tumor-infiltrating B lymphocytes as a driver tool ("GPS") for anti-tumor T cell CARs]. / A tumorban felhalmozódó B-limfociták genetikai információja, mint az immun T-sejt vezette tumorellenes "autó" (CAR, kiméra antigénreceptor) célba irányító "GPS"-e.

The rapidly growing field of gene therapy techniques to modify T cells with chimeric antigen receptors (CARs) for cancer care solutions, reached considerable achievements. However, there is an urgent need of reliable, well tolerable tumor-associated antigen specific antibodies. Tumor-infiltrating B (TIL-B) cell originated single chain Fv (scFv) gene regions could be selected with tumor specificity. DNA sequences of these antibody variable regions were subjects to get engineered into new CAR constructs. Our novel strategy harnesses tumor-infiltrating B cells' unique capacity to reveal highly tumor-associated disialylated glycosphingolipids (GD3 gangliosides). We used these human antibody fragments for generating GD3 ganglioside specific CAR gene constructs for potential usage in solid tumors.

[Minimally invasive breast surgery]. / Minimálisan invazív emlosebészet.

Due to the development in medical science and industrial technology, minimally invasive procedures have appeared in the surgery of benign and malignant breast diseases. In general , such interventions result in significantly reduced breast and chest wall scars, shorter hospitalization and less pain, but they require specific, expensive devices, longer surgical time compared to open surgery. Furthermore, indications or oncological safety have not been established yet. It is quite likely, that minimally invasive surgical procedures with high-tech devices - similar to other surgical subspecialties -, will gradually become popular and it may form part of routine breast surgery even. Vacuum-assisted core biopsy with a therapeutic indication is suitable for the removal of benign fibroadenomas leaving behind an almost invisible scar, while endoscopically assisted skin-sparing and nipple-sparing mastectomy, axillary staging and reconstruction with latissimus dorsi muscle flap are all feasible through the same short axillary incision. Endoscopic techniques are also suitable for the diagnostics and treatment of intracapsular complications of implant-based breast reconstructions (intracapsular fluid, implant rupture, capsular contracture) and for the biopsy of intracapsular lesions with uncertain pathology. Perception of the role of radiofrequency ablation of breast tumors requires further hands-on experience, but it is likely that it can serve as a replacement of surgical removal in a portion of primary tumors in the future due to the development in functional imaging and anticancer drugs. With the reduction of the price of ductoscopes routine examination of the ductal branch system, guided microdochectomy and targeted surgical removal of terminal ducto-lobular units or a "sick lobe" as an anatomical unit may become feasible. The paper presents the experience of the authors and provides a literature review, for the first time in Hungarian language on the subject. Orv. Hetil., 2014, 155(5), 162-169.

[The role of ductoscopy in the modern diagnostics and therapy of breast diseases]. / A duktoszkópia szerepe az emlobetegségek modern diagnosztikájában és kezelésében.

Mammary ductoscopy is a modern, minimally invasive procedure that enables direct, in vivo observation of the mammary ductal system, primarily by nipple discharge. The rapidly developing device is suitable for aimed biopsy for further cytological or molecular examinations. High-tech equipments facilitate polypectomy or laser vaporization of certain intraluminal lesions, and play an important role in the direct surgical excision of the duct or the so-called terminal duct-lobular unit. The above listed facilitate the early diagnosis of malignancies even before imaging could detect them, and the control of high risk patients. Ductoscopy can foster surgical removal of ductal in situ tumors as anatomical units, thus enabling the optimization of radicality of breast conserving surgeries. Authors give a detailed description of the surgical techniques, and provide a wide review of the literature, for the first time in the Hungarian language. Orv. Hetil., 2011, 152, 1284-1293.

[Histologic response after neoadjuvant therapy in rectal adenocarcinoma: own experience and review of the literature]. / Rectumcarcinomák neoadjuváns kezelést követo szövettani regressziójának vizsgálata: saját tapasztalatok és irodalmi áttekintés.

BACKGROUND: Neoadjuvant therapy has improved outcomes for patients with locally advanced rectal cancer. AIM AUTHORS: We studied the degree of histopathological regression after radiotherapy regimens commonly used in their clinical practice, and its effect on the pathology of by clinically founded T3-T4 rectal cancer. PATIENT AND METHODS: A total of 57 patients were investigated who underwent neoadjuvant radiotherapy or chemoradiotherapy--commonly used at our institute--for biopsy proven primary mid or lower third rectum adenocarcinoma between January and December 2004. The standard surgical treatment was anterior resection sec Dixon, Lloyd-Davies--and Hartmann's procedure, all with total mesorectal excision. RESULTS: The surgical specimens were examined by selected pathologists and a modified pathologic staging system the Rectal Cancer Regression Grade (RCRG) was used. (I) "good" response (n=12, 21%), (II) "median" response (n=25, 44%), (III) "poor" response (n=20, 35%). Significant tumor regression (RCRG I-II.) was seen in 37 patients (65%). Six patients (10.5%) have had complete pathological regression and 6 patients (10.5%) have had only microscopic foci of adenocarcinoma. CONCLUSION: Comparing of pretreatment and pathologic staging revealed that the depth of invasion was significantly downstaged. The authors also reviewed the literature of neoadjuvant therapy, total mesorectal excision, prognostic and predictive factors of rectal cancer.

[The role of sentinel lymph node biopsy in breast cancer non-invading-duct]. / Az orszemnyirokcsomó-biopszia jelentosége nem invazív duktális emlorákban.

INTRODUCTION AND AIMS: The clinical significance of sentinel lymph node biopsy for staging patients with ductal carcinoma in situ has not yet been solved. Determining the role of this method for the treatment of in situ ductal carcinoma has been the aim of this study. PATIENTS AND METHOD: Dual agent guided sentinel lymph node biopsy with preoperative lymphoscintigraphy was performed on 36 patients with breast ductal carcinoma in situ from January 2001 to March 2004 at the Department of General and Thoracic Surgery, National Institute of Oncology, Budapest. Completion axillary lymph node dissection was not performed routinely. The sentinel lymph nodes were histologically examined at 0.5-1 mm levels with routine hematoxylin and eosin staining. RESULTS: One patient has been excluded from the final analysis because of contralateral invasive breast cancer and simultaneous local recurrence in her medical history. Micro- or submicrometastases were found in 2 patients. If our patient number is completed with the 5 patients operated on for ductal carcinoma in situ during the period of our feasibility study (from December 1997 to March 2000) then the rate of patients with positive sentinel lymph node(s) will be 5% (2/40). All metastases were less than 2 mm in size. Metastases were found only in patients with high risk, extended ductal carcinoma in situ who finally underwent mastectomy. Completion axillary lymphadenectomy has not been performed even for patients with positive sentinel lymph node and no regional recurrence has yet been observed. DISCUSSION AND CONCLUSIONS: Our results corresponds well to the international ones. Performing sentinel lymph node biopsy for ductal carcinoma in situ of the breast is not recommended on the basis of the international and our own experiences. Sentinel lymph node biopsy is essential for patients undergoing mastectomy. In other cases when preoperative diagnostic studies do not verify invasion unequivocally we advise to perform sentinel lymph node biopsy (if necessary) after the final histological result of the excised breast specimen.

[The feasibility and the role of sentinel lymph node biopsy after neoadjuvant chemotherapy in breast cancer]. / Az ôrszemnyirokcsomó-biopszia alkalmazhatósága és jelentôsége neoadjuváns kemoterápia után emlôrákban.

INTRODUCTION AND AIMS: The feasibility, accuracy and clinical significance of sentinel lymph node biopsy for patients with breast cancer after neoadjuvant chemotherapy has not yet been determined. The aim of this study was to investigate these questions. PATIENTS AND METHOD: Dual agent-guided sentinel lymph node biopsy with preoperative lymphoscintigraphy was performed on 17 breast cancer patients after neoadjuvant chemotherapy at the Department of General and Thoracic Surgery, National Institute of Oncology, Budapest, from April 2004 to August 2005. Patients with clinically lymph node-negative breast cancer less than 3 cm in size after neoadjuvant chemotherapy were enrolled in the study. RESULTS: Lymphoscintigraphy showed no axillary lymphatic drainage in 7 patients (41%), and no sentinel lymph node could be identified during surgery in these patients. Axillary lymph nodes were histologically positive in 6 (86%) out of these 7 patients. Sentinel lymph node biopsy was successful in 10 patients (59%), and in 8 (80%) of them the sentinel lymph node proved to be positive pathologically. False negative sentinel lymph node biopsy did not occur. Axillary lymph node status was histologically positive in 14 (82%) out of the 17 patients. The predictable value of the clinical examination of the axilla after neoadjuvant chemotherapy, for the histological nodal status, was very low. DISCUSSION AND CONCLUSIONS: Our sentinel lymph node identification rate is lower than the published average in the literature. This difference can be explained by the differences in the indication for neoadjuvant chemotherapy. Our false negative rate (0%) is, however, significantly better than that of others. On the basis of international experiences sentinel lymph node biopsy after neoadjuvant chemotherapy is technically feasible, but its accuracy is not satisfactory and its clinical significance has not yet been determined. Our success rate is specifically low, which cannot be explained by the lack of practice. Taking the histologically very high axillary positive rate into consideration, sentinel lymph node biopsy has no clinical role in our practice after neoadjuvant chemotherapy.

[Leiomyosarcoma of the oesophagus: a case report]. / Nyelocso leiomyosarcomákról, egy operált esetünk kapcsán.

Authors treated a oesophageal sarcoma in a 46 year old female patient in July 1999. The tumor caused retrosternal pain and weight loss but did not disturb swallowing. After resecting the tumor, an intrathoracic oesophago-gastrostomy was performed. On the 18th postoperative day the patient went home without any complaints. Histologically the tumor proved to be a leiomyosarcoma. The patient has had no complaints in the past 6 years. On account of this case report the authors review the literature on this rare disease.

[Administration of clexane 4000 U/0.4 ml (40 mg) for preventing venous thromboembolic complications in patients undergoing surgery for malignant tumors. A follow-up study]. / A Clexane 4000 E/0.4 ml (40 mg) injekció alkalmazásának nyomonkövetése vénás thromboemboliás szövodmény profilaxisa során onkológiai mutéten átesett bete- gek esetében.

It has been evident since the first publication by Armand Trousseau (1865) that there is correlation between venous thromboembolism (VTE) and malignant tumors. This correlation has also been shown by recent studies. In patients with malignant tumors not only the incidence of VTE is higher but the course of VTE is more severe and relapses are more frequent. The procedures applied for the management of tumors (surgery, chemotherapy, radiotherapy, tamoxifen, central venous catheters) also increase the risk of VTE. It is generally accepted in Europe and in the U.S.A. (and also this constitutes the basis of the Hungarian guidelines) to group surgical patients by the risk of VTE by taking into account the characteristics of both the patients and the procedures. Recommendations also agree that for patients with medium or higher risk pharmacological prophylaxis is needed. Heparin or low molecular weight heparins in risk dependent doses and duration are administered most frequently. According to the recommendation of the American College of Chest Physicians (ACCP) patients with malignant tumors should receive thromboprophylaxis in accordance with their actual risk category and independent of the treatment they are receiving (surgery or chemotherapy). In a placebo controlled study with prolonged pharmacological thromboprophylaxis for patients with operated malignancies 60% VTE risk reduction could be reached and this risk reduction could also been observed after 3 months while the incidence of hemorrhagic complications was not significantly increased. Recent guidelines recommend 4 week long prophylaxis for patients with malignancies.

[Experience with radiofrequency ablation in the treatment of unresectable pelvic recurrence of rectal cancer]. / A radiofrekvenciás tumorablatió alkalmazásának lehetóségei recidív, kismedencei, irreszekábilis carcinomák kezelésében.

More than half of colorectal cancers are located in the rectum, and the number of such cancers is increasing. In Hungary colorectal cancers are diagnosed predominantly in advanced stages. In the last five years 736 patients with colorectal cancer were operated on at our Department, with the following stage distribution: Dukes A 10%, BI 10%, B2 31%, C 36% and D 13%. The local recurrence rate is decreasing since the introduction of total mesorectal excision and preoperative radiation. Effective treatment options are however poor for unresectable pelvic recurrences. Chemo- and radiotherapy have severe limitations in this advanced stage cancer. In recent years there are a few publications on the minimal-invasive radiofrequency tumour ablation (RFTA) technique, which is an effective treatment for primary and metastatic liver carcinomas and is a new palliative for the local treatment of pelvic recurrence. The aim of this study was to assess the response to treatment using ultrasound-guided radiofrequency ablation in two patients with unresectable pelvic recurrent rectal cancer.

[The similarities between the mechanism of wound healing and tumor development--literature review on the occasion of a patient with colonic adenocarcinoma metastasis in a dog-bite wound]. / A sebgyógyulás és a tumorprogresszió mechanizmusának közös lépései. Irodalmi áttekintés egy kutyaharapás sebében kialakult colonadenocarcinoma metasztázis esete kapcsán.

For more than a century, the role of wound healing in the growth of tumours has been implied based on observations in experimental and clinical studies. In the recent 10-15 years processes such as cell adhesion, angiogenesis, metastatic cascade and growth factors (fe: EGF, TGF-alpha, TGF-beta, IGF-1, IGF-II, PDGF) have been identified to play key roles in different stages of wound healing as well as in tumor progression. Experimental and clinical studies have established that wound healing creates a suitable environment favouring tumour growth and metastatic potential. Based on these observations, the clinical relevance of surgical procedures comes up, may playing a role in tumor recurrence after primary resection for cancer. This makes the opportunity of the development of completely new targeted approaches (antiangiogenic therapy, dormancy therapy) for the treatment of cancer. This review article is based on a case report of a 63-year-old female patient who had colonic adenocarcinoma metastasis of the hand after biting by a dog.

Subareolar injection of radioactive colloid for sentinel lymph node identification in breast cancer patients.

Sentinel lymph node biopsy (SLNB) is becoming the standard for staging the axilla in breast cancer patients in many institutions. The best method of injection is still questionable. The purpose of this study was to compare the results of SLNB using the peritumoral or the subareolar injection site. Between December 1997 and March 2000, we performed 100 SLNBs. Technecium-labeled colloidal human serum albumin was injected peritumorally (Group A, 31 patients; Group B, 31 patients) or subareolarly (Group C, 38 patients). Patent blue dye was given periareolarly (Group A) or peritumorally (Groups B and C). Preoperative lymphoscintigraphy was performed in all patients. SLNB was successful in 94 patients (94%). The identification rate improved from 80 per cent (first 25 patients) to 99 per cent (last 75). The subareolar injection of the colloid did not adversely influence the results of SLNB compared with the peritumoral injection (identification rate, 100% vs 97%; false negative rate, 6% vs 14%). The subareolar injection of colloid is a simple and at least as accurate technique as the peritumoral one. This technique can also improve the identification rate of SLNB for breast cancer patients.

[Change of attitude in the surgical treatment of lung metastases]. / Szemléletünk alakulása a tüdoáttétek sebészi kezelésében.

There has been no major change in surgical technique of lung metastasectomy till the last decades. In the latest years the variety of diagnostic tools and methods grew significantly. There are more chemotherapy possibilities and more palliative surgical techniques (VATS, ITP, RFTA) to treat certain types of lung metastases. Based on data of the International Registry of Lung Metastases the most important prognostic factor for metastasectomy is how radical the procedure was (clear resection margin). The more aggressive approach (even pneumonectomy for R0 resection) is becoming more popular.

[Radioguided excision of non-palpable breast lesions with simultaneous sentinel lymph node biopsy]. / Nem tapintható emlóelváltozások radioizotópos jelöléses eltávolítása egyidejú órszem nyirokcsomó-biopsziával.

As a result of mammographic screening nonpalpable breast lesions are found with increasing frequency. The surgical management of these lesions presents two main problems: (1) How to localize the lesion pre- and intraoperatively; (2) What should be the extension of the axillary dissection for malignant lesions. This study is aimed to evaluate the technical feasibility of radioguided excision of nonpalpable breast lesions and the possibility of performing simultaneous sentinel lymph node biopsy for malignant lesions. Radioguided breast excisions have been performed in our Department since November 2000. The basis of this method is that radioisotope labelled colloid is injected into the lesion under stereo tactic or sonographic guidance. A gamma-probe is used intraoperatively to locate the lesion and guide its excision. Dual agent guided technique is used for sentinel lymph node biopsy. Radioguided breast excision was performed on 85 patients (93 lesion) till February 2002. The localization and excision were successful in 84 patients (92 lesions) (99%). Multifocal lesions were successfully localized and excised in 7 patients. There were 61 parenchyma lesions (66%) and 31 microcalcifications (34%). Localization was performed under sonographic guidance in 44 patients (52%) and under stereotactic guidance in 40 patients (48%). There were 25 benign (27%) and 67 malignant (73%) lesions. Second surgical procedure (mastectomy) was needed in 4 patients (6.5%) because of histologically incomplete excision. Sentinel lymph node biopsy was performed in 50 patients. The biopsy was successful in 45 patients (90%). The sentinel lymph node was histologically positive in 2 patients (4.4%). Radioguided localization with the intraoperative use of a gamma-probe is an easy, rapid and highly accurate technique for removing nonpalpable breast lesions and allows simultaneous sentinel lymph node biopsy for malignant lesions.

[The role of surgery in the treatment of malignant melanoma]. / A sebészet szerepe a melanoma malignum kezelésében.

Surgical interventions have important role in the treatment of all stages of malignant melanoma. Surgery is the primary treatment of localized cutaneous melanoma. Excision of the primary tumor makes it possible to set up the histological diagnosis and to determine pathological prognostic factors. Appropriate surgical margin is important for local disease control. Sentinel lymph node biopsy with detailed histological examination has gained prominent importance for correct histological staging and for determining adjuvant oncological treatment. Surgery is the primary treatment of isolated regional metastases. Surgical methods also have a role in the palliative management of distant metastatic melanoma. In the present review the most important issues of the surgical treatment of malignant melanoma have been discussed in detail.

[Preoperative lymphoscintigraphy guided sentinel lymph node biopsy in malignant melanoma] / Preoperatív limfoszcintigráfiával végrehajtott sentinel nyirokcsomó mûtétek melanoma malignumban.

The authors present preliminary experience with preoperative sentinel lymph node biopsy carried out with lymphoscintigraphy in patients with malignant melanoma. PATIENTS AND METHODS: In the present study patients operated for primary cutaneous malignant melanoma of moderate and high severity were included. On the day of surgery isotope labelled colloid was injected intradermally around the tumor to indicate the lymphatics and to obtain basic information about the localization of the sentinel lymph node(s).During surgery the lymph node(s) previously visualized by the injection of patent-blue staining were detected with the aid of a gamma probe. Simultaneously, the excision of the primary tumor was extended. Histologically verified metastasis in the surgically removed lymph node(s) necessitated block dissection possibly within two weeks. RESULTS: The distribution of patients (19) according to tumor localisation: 2 - upper extremities; 9 - lower extremities; 2 - sacral region; 6 - trunk. Tumor thickness ranged from <1.5 mm (6 patients) to 1.5-3 mm (5 patients) and to >3 mm (8 patients). In two cases the identification of the lymph node has failed. Positive sentinel ymph nodes were detected in two patients. It is noteworthy that with one patient the sentinel lymph node was not regional but intransit. This study was aimed at the development of a suitable method. Further on we wish to try it in prospective randomized studies.