The introduction of sentinel lymph node biopsy (SLNB) has conveyed several new issues, such as the risk of false negativity, long-term consequences, the prognostic significance of micrometastases and whether ALND can be omitted in sentinel lymph node- (SLN) positive patients. Archived SLN specimens from 50 false negative patients and 107 true negative controls were serially sectioned and stained with immunohistochemistry. The detection rate of previously unknown metastases did not differ between the false and the true negative patients. The risk of false negativity was higher in patients with multifocal or hormone receptor-negative tumours, or if only one SLN was found. In a Swedish multicentre cohort, 2216 SLN-negative patients in whom ALND was omitted were followed up for a median of 65 months. The isolated axillary recurrence rate was only 1.0%, and the overall survival was high (93%). The survival of 3369 breast cancer patients (2383 node-negative (pN0), 107 isolated tumour cells (pN0(i+), 123 micrometastases (pN1mi) and 756 macrometastases (pN1)) was analysed. The 5-year cause-specific and event-free survival was worse for pN1mi and pN1 patients than for pN0 patients. There was no difference in survival between pN0(i+) and pN0 patients. Tumour and SLN characteristics in 869 SLN-positive patients were compared between those with and without non-SLN metastases, and the Tenon score was calculated. The risk of non-SLN metastases was higher in case of SLN macrometastases (compared with micrometastases), a high positive/total SLN ratio and Elston grade 3 tumours, and increased with increasing tumour size. The area under the curve (AUC) for the Tenon score was 0.65, and the test thus performed inadequately in this population. In conclusion, despite the risk of false negativity, SLNB with omission of ALND in SLN-negative patients appears to be safe even in the long term. The presence of micrometastases is of prognostic importance and should entail adjuvant treatment. The need for ALND in patients with SLN micro- and even macrometastases has been questioned, but the occurrence of non-SLN metastases is hard to predict, and strong evidence for the safe omission of ALND is lacking.

A linfocintilografia tem contribuído muito para a visualização da drenagem linfática e linfonodo sentinela (LNS) acometido e o estado patológico do LNS é considerado o fator prognóstico mais importante do melanoma. Os objetivos deste trabalho foram descrever os padrões de drenagem linfática em pacientes operados por melanoma, avaliar as características clínicas, demográficas e cirúrgicas destes pacientes, assim como a contribuição da Linfocintilografia na localização dos LNS com vistas à biópsia do mesmo. Foram avaliados 29 pacientes com melanoma operados, e a Linfocintilografia pré-operatória foi realizada após 60 minutos da injeção de Fitato marcado com Tecnécio 99m (99mTc), em 4 pontos cardeais a 1,0 cm da lesão. Encontramos maior acometimento por melanoma na população feminina e idosa, maioria em região de tronco posterior. A maioria (37,04%) drenou para 2 LNS/LNNS, 88,89% dos pacientes drenaram apenas para região ipsilateral à lesão, e 88,89% também drenaram apenas para um único território de drenagem. Encontramos padrões de drenagem linfática inesperados em melanoma localizados nas regiões de cabeça e pescoço, tronco anterior e tronco posterior. A técnica de linfocintilografia foi eficaz para evidenciar LNS/LNNS em 93,10% dos pacientes e dos 23 exames anatomopatológicos realizados, 6 (26,09%) apresentaram comprometimento metastático nos LNS. / Lymphoscintigraphy has contributed much to the visualization of lymphatic drainage and sentinel lymph node (SLN) and involved SLN pathological state is considered the most important prognostic factor of melanoma. Our objectives were to describe the lymphatic drainage patterns, to evaluate the clinical, demographic, and surgical data, as well as assessing the contribution of lymphoscintigraphy for SLN localization in a sample of patients operated for melanoma. We evaluated 29 patients operate with melanoma, and the Preoperative lymphoscintigraphy was performed after 60 minutes of injection of phytate labeled with Technetium 99m (99mTc) in 4 cardinal points to 1,0 cm of the lesion. We found greater involvement by melanoma in the elderly female population and, most located in the posterior trunk region. The majority (37.04%) drained for 2 SLN/ SLNN, 88.89% of the patients drained only to the ipsilateral region, and also 88,89% had lymphatic drainage to only a single drainage area. We found unexpected lymphatic drainage patterns in melanoma localized in head and neck, trunk, anterior and posterior trunk. The technique of lymphoscintigraphy was effective to show SLN/SLNN in 93.10% of patients, and from 23 pathological examinations performed, 6 (26.09%) exhibited metastatic involvement in the SLN.

<p>The most important prognostic factor in breast cancer is the axillary lymph node status. The sentinel node biopsy (SNB) is reported to stage the axilla with an accuracy > 95 % in early breast cancer. Tumour-related perturbation of T-cell function has been observed in patients with malignancies, including breast cancer. The down-regulation of the important T-cell activation molecules CD3-ζ and CD28 may cause T-cell dysfunction, anergy, tolerance and deletion.</p><p>The expression of CD3-ζ and CD28 was evaluated in 25 sentinel node biopsies. The most pronounced down-regulation was seen in the paracortical area, where the best agreement between both parameters was observed. CD28 expression was significantly more suppressed in CD4+ than in CD8+ T-cells.</p><p>From the Swedish sentinel node database, 109 patients with breast cancer > 3 cm planned for both SNB and a subsequent axillary dissection were identified. The false negative rate (FNR) was 12.5%. Thirteen cases of tumour multifocality were detected on postoperative pathology. The FNR in this subgroup was higher (30.8%) than in patients with unifocal disease (7.8%; P = 0.012).</p><p>From the Swedish SNB multicentre cohort trial, 2246 sentinel node-negative patients who had not undergone further axillary surgery were selected for analysis. After a median follow-up time of 37 months (range 0-75), 13 isolated axillary recurrences (13/2246; 0.6%) were found. In another 14 cases, local or distant failure preceded or coincided with axillary relapse (27/2246; 1.2%). </p><p>In conclusion, the immunological analysis of the sentinel node might provide valuable prognostic information and aid selection of patients for immunotherapy. SNB is encouraged in breast cancer larger than 3 cm, if no multifocal growth pattern is present. The axillary recurrence rate after a negative SNB in Sweden is in accordance with international figures. However, a longer follow-up is mandatory before the true failure rate of the SNB can be determined.</p>

The most important prognostic factor in breast cancer is the axillary lymph node status. The sentinel node biopsy (SNB) is reported to stage the axilla with an accuracy &gt; 95 % in early breast cancer. Tumour-related perturbation of T-cell function has been observed in patients with malignancies, including breast cancer. The down-regulation of the important T-cell activation molecules CD3-ζ and CD28 may cause T-cell dysfunction, anergy, tolerance and deletion. The expression of CD3-ζ and CD28 was evaluated in 25 sentinel node biopsies. The most pronounced down-regulation was seen in the paracortical area, where the best agreement between both parameters was observed. CD28 expression was significantly more suppressed in CD4+ than in CD8+ T-cells. From the Swedish sentinel node database, 109 patients with breast cancer &gt; 3 cm planned for both SNB and a subsequent axillary dissection were identified. The false negative rate (FNR) was 12.5%. Thirteen cases of tumour multifocality were detected on postoperative pathology. The FNR in this subgroup was higher (30.8%) than in patients with unifocal disease (7.8%; P = 0.012). From the Swedish SNB multicentre cohort trial, 2246 sentinel node-negative patients who had not undergone further axillary surgery were selected for analysis. After a median follow-up time of 37 months (range 0-75), 13 isolated axillary recurrences (13/2246; 0.6%) were found. In another 14 cases, local or distant failure preceded or coincided with axillary relapse (27/2246; 1.2%). In conclusion, the immunological analysis of the sentinel node might provide valuable prognostic information and aid selection of patients for immunotherapy. SNB is encouraged in breast cancer larger than 3 cm, if no multifocal growth pattern is present. The axillary recurrence rate after a negative SNB in Sweden is in accordance with international figures. However, a longer follow-up is mandatory before the true failure rate of the SNB can be determined.

La extensión linfática es el factor pronóstico más importante en el momento del diagnóstico del cáncer de mama. La Biopsia Selectiva del Ganglio Centinela (BSGC) es el procedimiento actualmente indicado para la estadificación regional de las pacientes con cáncer de mama precoz. La BSGC permite una estadificación más exacta que la linfadenectomía axilar tradicional (LDNA), gracias a la identificación de drenajes extra-axilares y de micrometástasis en el GC, de modo que el manejo terapeútico posterior de las pacientes ha de ser más acertado y la evolución clínica mejor. El objetivo de este trabajo es evaluar la eficiencia y seguridad de la aplicación clínica de la BSGC en las pacientes con cáncer de mama, perfilando la metodología óptima en cuanto al radiotrazador y la vía de administración, y analizando la indicación en situaciones clínicas especiales: carcinoma in situ, múltiple, o cirugía mamaria previa. Se estudian los primeros 600 procedimientos de BSGC realizados en fase clínica, a 591 pacientes con cáncer de mama estadio I y II. La eficiencia de detección gammagráfica e intraoperatoria del GC fue 94,5% y 90,5% respectivamente. El procedimiento de BSGC evita una LDNA innecesaria en el 68,9% de los pacientes, que presentan GC benigno. Después de un seguimiento medio de 47,4 meses, ha aparecido una única recaída axilar en las pacientes con GC benigno (tasa 0,24%). Este aspecto probablemente se relaciona con la exéresis sistemática de otros ganglios “sospechosos”. El 8,3 % de los pacientes incluidos han sido re-estadificados por la identificación de micrometástasis en el GC, hallando otros ganglios positivos en la LDNA en el 12,1%. La tasa de enfermedad es 8,1% en estas pacientes. La eficiencia de detección del GC no ha diferido significativamente al comparar radiotrazadores de distinto tamaño (coloide de estaño vs sulfuro de renio) y diferentes vías de administración (profunda, periareolar, combinada), aunque se debe considerar la fuerte dependencia entre ambos aspectos técnicos. Los drenajes extra-axilares son más frecuentes cuando se emplea un radiocoloide de menor tamaño. El análisis histopatológico final revela invasión en el 45,6% de los pacientes incluidos con el diagnóstico inicial de carcinoma in situ. El porcentaje de afectación del GC en estos pacientes es 6,9%. La tasa de identificación del GC en el carcinoma múltiple de mama es 95,5% en la gammagrafía y 92,1% en la detección intraoperatoria; 93,7% y 87,5% respectivamente en los tumores multicéntricos, en los que se observa además mayor número de GC por paciente y de GC extra-axilares. La eficiencia de detección gammagráfica y quirúrgica del GC en las pacientes con cirugía mamaria previa (inyección superficial) es 92,3% y 82% respectivamente. El número de GC por paciente y el porcentaje de drenajes extra-axilares son significativamente elevados, sobre todo cuando existe algún antecedente quirúrgico extenso antiguo (cirugía conservadora) y/o localizado en el CSE. En conclusión, la BSGC es una técnica eficaz, segura y fiable que permite una estadificación exacta de los pacientes y contribuye al control locorregional de la enfermedad. La inyección periareolar se confirma como método seguro y altamente eficaz, aún cuando se emplean radiocoloides de tamaño grande. El procedimiento de BSGC está indicado en cáncer in situ de alto riesgo o con indicación de mastectomía, y puede realizarse con seguridad en tumores múltiples, incluso multicéntricos, y en pacientes con antecedentes quirúrgicos, aunque sean relativamente extensos o afecten al CSE. En todas estas circunstancias es apropiada también la administración periareolar del radiocoloide. / Lymphatic status is the most important prognostic factor in breast cancer. Sentinel lymph node biopsy (SLNB) is the current elective procedure for regional staging in patients with early breast cancer. SLNB allows more exact staging than traditional axillary lymphadenectomy (ALDN), on the basis of the identification of extra-axillary drainages and sentinel node (SN) micrometastasis, what results in more appropriate adjuvant theraphy and better outcome of the patients. The aim of this study was to establish the efficiency and safety of SLNB in its clinical application to breast cancer patients, as well as the optimal methodology (particle size of the colloid, site of injection) and controversial indications (intraductal carcinoma, multifocal/multicentric breast cancer, patients with previous breast surgery). This study included the first 600 SLNB procedures, performed in 591 patients with breast cancer, I and II stage. The scintigraphic and intraoperative detection rate of SN were respectively 94.5% and 90.5%.The procedure avoided unnecesary ALDN in 68.9% of the patients, those without SN involvement. After a median follow-up of 47.4 months, the incidence of axillary recurrence after tumor negative SN is low (0.24%), probably related to the systematic removal of “suspicious” nodes during the surgical act. The identification of micrometastasis in SN implied the upstaging of 8.3% of patients included. The subsequent ALDN performed showed non-sentinel node metastasis in 12.1%. Disease rate in these patients is 8.1%. The efficiency of SN detection did not varied significantly when comparing radiotracers of different size (stannous fluorid vs rhenium sulphide colloid), or different sites of injection (deep, areolar or combined). However, the strong correlation between these two technical aspects must be taken into account. Extra-axillary drainages are more frequent when a smaller radiotracer is used. Final pathologic diagnosis revealed invasion in 45.6% of patients included with the initial diagnosis of ductal carcinoma in situ. The percentage of positive SN was 6.9% in these patients. The identification rate of SN in patients with multiple breast cancer was 93.7% in lymphoscintigraphy and 92.1% in surgical detection. These percentages were respectively 93.7% and 87.5% in multicentric cases, in that higher number of SN per patient and extra-axillary nodes were found. Scintigraphic and surgical detection rate in patients with previous breast surgery was 92.3% and 82%. The number of SN per patient and the percentage of extra-axillary drainages were higher, especially after wide ancient surgery (conservative) in outer-upper quadrant. In conclusion, SLNB is an efficient, safe and accurate procedure. It achieves an exact staging of the patients, and contributes to local control of the disease. Periareolar injection is a reliable method, even though large colloids are used. SLNB is indicated in high risk ductal carcinoma in situ or programmed to mastectomy, multiple breast cancer, multicentric included, and breast cancer in patients with prior breast surgery, even though wide or external. In these clinical circumstances, the periareolar injection is also appropiate.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2007. / Härtill 4 uppsatser.

A linfocintilografia tem contribuído muito para a visualização da drenagem linfática e linfonodo sentinela (LNS) acometido e o estado patológico do LNS é considerado o fator prognóstico mais importante do melanoma. Os objetivos deste trabalho foram descrever os padrões de drenagem linfática em pacientes operados por melanoma, avaliar as características clínicas, demográficas e cirúrgicas destes pacientes, assim como a contribuição da Linfocintilografia na localização dos LNS com vistas à biópsia do mesmo. Foram avaliados 29 pacientes com melanoma operados, e a Linfocintilografia pré-operatória foi realizada após 60 minutos da injeção de Fitato marcado com Tecnécio 99m (99mTc), em 4 pontos cardeais a 1,0 cm da lesão. Encontramos maior acometimento por melanoma na população feminina e idosa, maioria em região de tronco posterior. A maioria (37,04%) drenou para 2 LNS/LNNS, 88,89% dos pacientes drenaram apenas para região ipsilateral à lesão, e 88,89% também drenaram apenas para um único território de drenagem. Encontramos padrões de drenagem linfática inesperados em melanoma localizados nas regiões de cabeça e pescoço, tronco anterior e tronco posterior. A técnica de linfocintilografia foi eficaz para evidenciar LNS/LNNS em 93,10% dos pacientes e dos 23 exames anatomopatológicos realizados, 6 (26,09%) apresentaram comprometimento metastático nos LNS. / Lymphoscintigraphy has contributed much to the visualization of lymphatic drainage and sentinel lymph node (SLN) and involved SLN pathological state is considered the most important prognostic factor of melanoma. Our objectives were to describe the lymphatic drainage patterns, to evaluate the clinical, demographic, and surgical data, as well as assessing the contribution of lymphoscintigraphy for SLN localization in a sample of patients operated for melanoma. We evaluated 29 patients operate with melanoma, and the Preoperative lymphoscintigraphy was performed after 60 minutes of injection of phytate labeled with Technetium 99m (99mTc) in 4 cardinal points to 1,0 cm of the lesion. We found greater involvement by melanoma in the elderly female population and, most located in the posterior trunk region. The majority (37.04%) drained for 2 SLN/ SLNN, 88.89% of the patients drained only to the ipsilateral region, and also 88,89% had lymphatic drainage to only a single drainage area. We found unexpected lymphatic drainage patterns in melanoma localized in head and neck, trunk, anterior and posterior trunk. The technique of lymphoscintigraphy was effective to show SLN/SLNN in 93.10% of patients, and from 23 pathological examinations performed, 6 (26.09%) exhibited metastatic involvement in the SLN.

<p>This thesis deals with recognition and elimination of tumours by T lymphocytes and their use in adoptive immunotherapy.</p><p>The first tumour-draining lymph node; the sentinel node, is identified by peritumoural injection of a tracer. This is the hypothesised location for the activation of tumour-reactive lymphocytes. Accordingly, proliferation and IFN-γ production in response to autologous tumour extract was detected in sentinel nodes from patients with colon and urinary bladder cancer. Reactivity in metastatic nodes was generally lower or absent, but the non-responsiveness could be subdued in long-term cultures by addition of tumour antigen and IL-2. A novel padlock-probe based method was developed for measuring the T cell receptor Vβ repertoire. Common Vβ gene expansions were detected in tumour-infiltrating lymphocytes and sentinel nodes. Thus, tumour antigens are recognised in sentinel nodes by Th1 lymphocytes, resulting in a clonally expanded cell population that can be further propagated <i>ex vivo</i>.</p><p>Regulatory T cells (Tregs) may contribute to tumour-induced immunosuppression. Immunohistochemical stainings against the pan-T cell marker CD3 and Treg marker FOXP3 was performed on tumour tissue from 20 historical urinary bladder cancer patients. The ratio of FOXP3⁺ to CD3⁺ cells was lower in patients alive 7 years post-cystectomy, suggesting that Tregs in bladder cancer have prognostic implications.</p><p>Lymphocytes were isolated from sentinel nodes from sixteen patients with advanced or high-risk colon cancer. <i>In vitro</i> expansion with addition of autologous tumour extract and IL-2 mainly promoted the outgrowth of CD4⁺ Th1 lymphocytes, which were safely re-transfused to the patients. Four patients responded with complete tumour regression. Survival time in the Dukes’ D patients was significantly increased compared with conventionally treated controls (2.6 versus 0.8 years; p=0.048).</p><p>In conclusion, human solid tumours are recognised in sentinel nodes and <i>in vitro</i> expanded sentinel node-acquired CD4⁺ T lymphocytes seem useful in the treatment of patients with disseminated cancer.</p>

This thesis deals with recognition and elimination of tumours by T lymphocytes and their use in adoptive immunotherapy. The first tumour-draining lymph node; the sentinel node, is identified by peritumoural injection of a tracer. This is the hypothesised location for the activation of tumour-reactive lymphocytes. Accordingly, proliferation and IFN-γ production in response to autologous tumour extract was detected in sentinel nodes from patients with colon and urinary bladder cancer. Reactivity in metastatic nodes was generally lower or absent, but the non-responsiveness could be subdued in long-term cultures by addition of tumour antigen and IL-2. A novel padlock-probe based method was developed for measuring the T cell receptor Vβ repertoire. Common Vβ gene expansions were detected in tumour-infiltrating lymphocytes and sentinel nodes. Thus, tumour antigens are recognised in sentinel nodes by Th1 lymphocytes, resulting in a clonally expanded cell population that can be further propagated ex vivo. Regulatory T cells (Tregs) may contribute to tumour-induced immunosuppression. Immunohistochemical stainings against the pan-T cell marker CD3 and Treg marker FOXP3 was performed on tumour tissue from 20 historical urinary bladder cancer patients. The ratio of FOXP3+ to CD3+ cells was lower in patients alive 7 years post-cystectomy, suggesting that Tregs in bladder cancer have prognostic implications. Lymphocytes were isolated from sentinel nodes from sixteen patients with advanced or high-risk colon cancer. In vitro expansion with addition of autologous tumour extract and IL-2 mainly promoted the outgrowth of CD4+ Th1 lymphocytes, which were safely re-transfused to the patients. Four patients responded with complete tumour regression. Survival time in the Dukes’ D patients was significantly increased compared with conventionally treated controls (2.6 versus 0.8 years; p=0.048). In conclusion, human solid tumours are recognised in sentinel nodes and in vitro expanded sentinel node-acquired CD4+ T lymphocytes seem useful in the treatment of patients with disseminated cancer.

In den letzten Jahren wurde die Sentinel Node Biopsie in Studien zur Diagnostik und Behandlung des N0-Halses als Stagingverfahren untersucht. Ziel der vorliegenden Studie war der Vergleich der postoperativen Lebensqualität und des funktionellen Status bei Patienten mit oralen und oropharyngealen Karzinomen, die sich einer Sentinelbiopsie, bzw. einer selektiven Neck Dissection bei N0-Hals unterzogen haben. 57 Patienten mit primär operativ behandeltem Karzinom wurden im Rahmen der Studie untersucht, 29 nach Sentinel Node Biopsie und 28 nach selektiver Neck Dissection. Es wurde die postoperative gesundheitsbezogene (EORTC QLQ-C30) und krankheitsspezifische Lebensqualität (EORTC QLQ-H&N35) erfasst. Patientenbezogene psychosoziale Variablen wurden anhand der „Hospital Anxiety and Depression Scale“ (HADS) und der Kurzform des „Progredienzangst-Fragebogens“ (PA-F-KF) ermittelt. Die Erhebung des klinischen Gesundheitszustandes erfolgte unter besonderer Berücksichtigung des funktionellen Status nach zervikaler Lymphknotendissektion. Bezüglich der gesundheitsbezogenen Lebensqualität sowie der Angst und Depression existierten keine signifikanten Unterschiede. Hinsichtlich der krankheitsspezifischen Lebensqualität zeigte sich, dass die Patienten nach Sentinel Node Biopsie weniger Schluckbeschwerden angaben (p=0,037). Ebenso zeigte sich bei der klinischen Untersuchung eine geringere Beeinträchtigung durch die zervikalen Narben. Die Sentinel Node Biopsie bietet, neben dem geringeren operativen Aufwand, der schnelleren Rekonvaleszenz und der geringeren funktionellen Minderung der Patienten, eine tendenzielle Verbesserung der krankheitsspezifischen Lebensqualität. Als Grund für die subjektiv weniger beeinträchtigte Schluckfunktion kann die fehlende Präparation der suprahyoidalen Muskulatur und der entsprechend innervierenden Nerven in Erwägung gezogen werden.