Evaluation of tumor vascularization and microenvironment in follicular lymphoma


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AIM: To characterize the degree of follicular lymphoma (FL) vascularization and microenvironment by immunohistochemical studies (IHCS) of lymph node biopsy paraffin-embedded sections in 2 different disease pattern groups/MATERIAL AND METHODS: The investigation included 59 patients: 39 (67%) women and 20 (33%) men whose age was 27 to 83 years (median age 53 years) treated at the Hematology Research Center, Ministry of Health of the Russian Federation (n=49), and the N.N. Blokhin Russian Cancer Research Center, Russian Academy of Medical Sciences (n=10), in April 2001 to May 2011. In accordance with the clinical features of the disease, the authors identified 2 patient groups: 1) 31 patients with the good results of FL treatment and 2) 28 patients with its poo/RESULTS: IHCS was performed on lymph node tumor biopsy paraffin-embedded sections prior to treatment using antibodies to CD34, D2-40, CD68, and granzyme B. Morphometric analysis was made applying microscopy and a Leica ×400 digital camera. The images of histological specimens were processed by the computer program VideoTesT-Morphology 5.2: the specific vessel area (%) in relation with tumor tissue was estimated under visual guidance of an investigator. Cytotoxic lymphocytes (CTL) and macrophages were quantitatively characterized using 1 mm2 of tumor tissue (12 fields of vision with the objective lens magnifying ×400). Immunohistochemical specimens to be examined were chosen randomly, by using the random number table/RESULTS: In Group 2, the specific area of blood vessels was statistically significantly higher than in Group 1: 0.04% (95% confidence interval (CI), 0.03 to 0.05%) versus 0.02% (95% CI, 0.01 to 0.03%; p=0.05). In Group 2, that of lymphatic vessels was significantly higher than in Group 1: 0.06% (95% CI, 0.04 to 0.07%) versus 0.03% (95% CI, 0.01 to 4%; p=0.03). With a nodular diffuse growth, Group 2 showed a significantly more CD68-positive macrophages than did Group 1: 800 (95% CI, 380 to 1222) versus 79 (95% CI, 10 to 566; р=0.01). In Group 1, the count of CTL was statistically significantly (p=0.05) higher than in Group 2 in both the nodule (with a nodular growth pattern: 14 (5-27) versus 5 (1-11)) and the internodular space (with a nodular growth pattern: 158 (118-410) versus 35 (5-287) and with a nodular diffuse growth pattern: 126 (102-360) versus 35 (3-120))/CONCLUSION: Increased tumor vascularization (estimated by the specific density of tumor vasculature) and a pronounced macrophageal reaction are associated with the poor outcomes of FL; the marked cytotoxic component in tumor tissue is linked to the favorable outcomes of the disease.

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Оценка васкуляризации и микроокружения опухолевой ткани при фолликулярной лимфоме. - Резюме. Цель исследования. Оценить васкуляризацию и микроокружение опухолевой ткани при фолликулярной лимфоме (ФЛ) на основе иммуногистохимического исследования (ИГХИ) на срезах с парафиновых блоков биоптатов лимфатических узлов в сравнительном исследовании 2 групп пациентов, отличающихся исходами заболевания. Материалы и методы. В исследование включили 59 больных (39 (67%) женщин и 20 (33%) мужчин) в возрасте от 27 до 83 лет (медиана 53 года), проходивших лечение с апреля 2001 г. по май 2011 г. в Гематологическом научном центре МЗ РФ (n=49) и в Российском онкологическом научном центре им. Н.Н. Блохина РАМН (n=10). В соответствии с особенностями клинического течения заболевания выделены 2 группы пациентов: 1-я (n=31) - с хорошими результатами лечения ФЛ и 2-я (n=28) - с неблагоприятными исходами. ИГХИ выполнено на срезах с парафиновых блоков биоптатов опухолевых лимфатических узлов до лечения с использованием антител к CD34, D2-40, CD68 и granzyme B. Морфометрический анализ проведен с использованием микроскопии и цифровой камеры Leica (об. ×400). Фотографии гистологических препаратов обработаны с помощью компьютерной программы "ВидеоТесТ-Морфология 5.2": оценена удельная площадь сосудов (в процентах) по отношению к опухолевой ткани при визуальном контроле исследователя. Количественная характеристика цитотоксических лимфоцитов (ЦТЛ) и макрофагов проводилась на 1 мм2 опухолевой ткани (12 полей зрения при увеличении объектива ×400). Выбор иммуногистохимических препаратов для исследования проводили случайным образом (с помощью таблицы случайных чисел). Результаты. Удельная площадь кровеносных сосудов во 2-й группе оказалась статистически значимо больше, чем в 1-й: 0,04% (при 95% доверительном интервале - ДИ от 0,03 до 0,05%) против 0,02% (при 95% ДИ от 0,01 до 0,03%; р=0,05). Удельная площадь лимфатических сосудов во 2-й группе значимо больше, чем в 1-й: 0,06% (при 95% ДИ от 0,04 до 0,07%) против 0,03% (при 95% ДИ от 0,01 до 4%; p=0,03). При нодулярно-диффузном характере роста во 2-й группе количество CD68-позитивных макрофагов оказалось значимо больше, чем в 1-й: 800 (при 95% ДИ от 380 до 1222) против 79 (при 95% ДИ от 10 до 566; р=0,01). Количество ЦТЛ в 1-й группе статистически значимо (р=0,05) больше, чем во 2-й, как в нодулярном (при нодулярном росте опухоли: 14 (5-27) против 5 (1-11)), так и в интернодулярном пространстве (при нодулярном характере роста опухоли: 158 (118-410) против 35 (5-287) и при нодулярно-диффузном характере роста: 126 (102-360) против 35 (3-120)). Заключение. Повышенная васкуляризация опухоли (оцениваемая удельной плотностью сосудистой сети в опухолевой ткани) и выраженная макрофагальная реакция ассоциированы с неблагоприятными исходами фолликулярной лимфомы; выраженный цитотоксический компонент в опухолевой ткани ассоциирован с благоприятными исходами заболевания.
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