Evaluation of tumor vascularization and microenvironment in follicular lymphoma

Nesterova E.S.; Kravchenko S.K.; Gemdzhyan E.G.; Osmanov E.A.; Kovrigina A.M.

doi:https://doi.org/

Close metadata

Recommended articles:

Characteristics of the immune microenvironment of endometrial cancer depending on the MMR status of the tumor. Russian Journal of Archive of Pathology. 2025;(5):11-19

Spiny mice (Acomys): a novel and perspective model for research of heart regeneration and development of regenerative technologies. Russian Cardiology Bulletin. 2025;(4):10-17

Role of lipopolysaccharide in the development of inflammation in placental tissue in preeclampsia. Russian Journal of Human Reproduction. 2026;(2):97-103

Abstract:

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. Subjects 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 poor 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.

Keywords:

follicular lymphoma

blood vessels

lymphatic vessels

macrophages

cytotoxic lymphocytes

exploratory data analysis

multivariate statistical analysis

Authors:

Nesterova E.S.

Gematologicheskiĭ nauchnyĭ tsentr Minzdrava Rossii

Kravchenko S.K.

Gematologicheskiĭ nauchnyĭ tsentr Minzdrava Rossii

Gemdzhyan E.G.

National Research Center for Hematology, Moscow, Russia

Osmanov E.A.

Rossiĭskiĭ onkologicheskiĭ nauchnyĭ tsentr im. akad. N.N. Blokhina RAMN, Moskva

Kovrigina A.M.

Nauchno-klinicheskoe otdelenie gematologicheskoĭ khirurgii Gematologicheskogo nauchnogo tsentra Minzdrava RF, Moskva

Close metadata

References:

Morton L.M., Wang S.S., Devesa S.S. et al. Lymphoma incidence patterns by WHO subtype in the United States, 1992-2001. Blood 2006; 107 (1): 265-276.
A clinical evaluation of the International Lymphoma Study Group classification of non-Hodgkin's lymphoma. The Non-Hodgkin's Lymphoma Classification Project. Blood. 1997; 89 (11): 3909-3918.
Rohatiner A.Z., Nadler L., Davies A.J. et al. Myeloablative therapy with autologous bone marrow transplantation for follicular lymphoma at the time of second or subsequent remission: long-term follow-up. J Clin Oncol 2007; 25 (18): 2554-2559.
van Oers M.H., Van Glabbeke M., Giurgea L. et al. Rituximab maintenance treatment of relapsed/resistant follicular non-Hodgkin's lymphoma: long-term outcome of the EORTC 20981 phase III randomized intergroup study. J Clin Oncol 2010; 28 (17): 2853-2858.
Schouten H.C., Qian W., Kvaloy S. et al. High-dose therapy improves progression-free survival and survival in relapsed follicular non-Hodgkin's lymphoma: results from the randomized European CUP trial. J Clin Oncol 2003; 21 (21): 3918-3927.
Hazar B., Paydas S., Zorludemir S. et al. Prognostic significance of microvessel density and vascular endothelial growth factor (VEGF) expression in non-Hodgkin's lymphoma. Leuk Lymphoma 2003; 44: 2089-2093.
Tzankov A., Heiss S., Ebner S. Angiogenesis in nodal B cell lymphomas: a high throughput study. J Clin Pathol 2007; 60: 476-482.
Ferrara N., Winer J., Burton T. et al. Expression of vascular endothelial growth factor does not promote transformation but confers a growth advantage in vivo to Chinese hamster ovary cells. J Clin Invest 1993; 91: 160-170.
Zhang H.T., Craft P., Scott P.A. et al. Enhancement of tumor growth and vascular density by transfection of vascular endothelial cell growth factor into MCF-7 human breast carcinoma cells. J Nat Cancer Inst 1995; 87: 213-219.
Murrell J.C., Lidstrom M.E., Holmes A.J., Costello A. Evidence that particulate methane monooxygenase and ammonia monooxygenase may be evolutionarily related. FEMS Microbiol Lett 1995; 132 (3): 203-208.
Yamaguchi H., Takagi J., Miyamae T. et al. Milk fat globule EGF factor 8 in the serum of human patients of systemic lupus erythematosus. J Leukoc Biol 2008; 83 (5): 1300-1307.
Barleon B., Sozzani S., Zhou D. et al. Migration of human monocytes in response to vascular endothelial growth factor (VEGF) is mediated via the VEGF receptor flt-1 Blood 1996; 87: 3336-3343.
Hiratsuka S., Minowa O., Kuno J. et al. Flt-1 lacking the tyrosine kinase domain is sufficient for normal development and angiogenesis in mice. Proc Natl Acad Sci USA 1998; 95: 9349-9354.
Yang Z.F., Poon R.T., Luo Y. et al. Up-regulation of vascular endothelial growth factor (VEGF) in small-for-size liver grafts enhances macrophage activities through VEGF receptor 2-dependent pathway. J Immunol 2004; 173: 2507-2515.
Skobe M., Hamberg L.M., Hawighorst T. et al. Concurrent induction of lymphangiogenesis, angiogenesis, and macrophage recruitment by vascular endothelial growth factor-C in melanoma. Am J Pathol 2001; 159: 893-903.
Pepper M.S. Lymphangiogenesis and Tumor Metastasis: Myth or Reality? Clin Cancer Res 2001; 7 (3): 462-468.
Schneider M., Othman-Hassan K., Christ B., Wilting J. Lymphangioblasts in the avian wing bud. Dev Dyn 1999; 216: 311-319.
Takahashi T., Kalka C., Masuda D. et al. Ischemia- and cytokine-induced mobilization of bone marrow-derived endothelial progenitor cells for neovascularization. Nat Med 1999; 5: 434-438.
Gill M., Dias K., Hattori M.L. et al. Vascular trauma induces rapid but transient mobilization of VEGFR2(+) AC133(-) endothelial precursor cells. Circ Res 2001; 88: 167-174.
Dengjel J., Kratchmarova I., Blagoev B. Receptor tyrosine kinase signaling: a view from quantitative proteomics. Mol Biosyst 2009; 5 (10): 1112-1121.
Folkman J., D'Amore P.A. Blood vessel formation: what is its molecular basis? Cell 1996; 87: 1153-1155.
Holmes K., Owain R.L., Angharad M.T. Cross M.J. Vascular endothelial growth factor receptor-2: Structure, function, intracellular signalling and therapeutic inhibition. Cellular Signalling 2007; 19 (10): 2003-2012.
Wey J.S., Fan F., Gray M.J. et al. Vascular endothelial growth factor receptor-1 promotes migration and invasion in pancreatic carcinoma cell lines. Cancer 2005; 104: 427-438.
Koster A., Raemaekers J.M. Angiogenesis in malignant lymphoma. Curr Opin Oncol 2005; 17: 611-616.
Foss H.D., Araujo I., Demel G. et al. Expression of vascular endothelial growth factor in lymphomas and Castleman's disease. J Pathol 1997; 183: 44-50.
Ribatti D., Vacca A., Bertossi M. et al. Angiogenesis induced by B-cell non-Hodgkin's lymphomas. Lack of correlation with tumor malignancy and immunologic phenotype. Anticancer Res 1990; 10: 401-406.
Stewart M., Talks K., Leek R. et al. Expression of angiogenic factors and hypoxia inducible factors HIF 1, HIF 2 and CA IX in non-Hodgkin's lymphoma. Histopathology 2002; 40: 253-260.
Jørgensen J.M., Sørensen F.B., Bendix K. Expression level, tissue distribution pattern, and prognostic impact of vascular endothelial growth factors VEGF and VEGF-C and their receptors Flt-1, KDR, and Flt-4 in different subtypes of non-Hodgkin lymphomas. Leuk Lymphoma 2009; 50 (10): 1647-1660.
Ho C.L., Sheu L.F., Li C.Y. Immunohistochemical expression of angiogenic cytokines and their receptors in reactive benign lymph nodes and non-Hodgkin lymphoma. Ann Diagn Pathol 2003; 7 (1): 1-8.
Gately S., Li W.W. Multiple roles of COX-2 in tumor angiogenesis: a target for antiangiogenic therapy. Semin Oncol 2004; 31 (Suppl 7): 2-11.
Hoper M.M., Voelkel N.F., Bates T.O. et al. Prostaglandins induce vascular endothelial growth factor in a human monocytic cell line and rat lungs via cAMP. Am J Respir Cell Mol Biol 1997; 17: 748-756.
Bellamy W.T., Richter L., Frutiger Y. et al. Expression of vascular endothelial growth factor and its receptors in hematopoietic malignancies. Cancer Res 1999; 59: 728-733.
Bono P., Teerenhovi L., Joensuu H. Elevated serum endostatin is associated with poor outcome in patients with non-Hodgkin lymphoma. Cancer 2003; 97: 2767-2775.
Pedersen L.M., Klausen T.W., Davidsen U.H. et al. Early changes in serum IL-6 and VEGF levels predict clinical outcome following first-line therapy in aggressive non-Hodgkin's lymphoma. Ann Hematol 2005; 84: 510-516.
Potti A., Ganti A.K., Kargas S. et al. Immunohistochemical detection of C-kit (CD117) and vascular endothelial growth factor (VEGF) overexpression in mantle cell lymphoma. Anticancer Res 2002; 22: 2899-2901.
Salven P., Orpana A., Teerenhovi L. et al. Simultaneous elevation in the serum concentrations of the angiogenic growth factors VEGF and bFGF is an independent predictor of poor prognosis in non-Hodgkin lymphoma: a singleinstitution study of 200 patients. Blood 2000; 96: 3712-3718.
Vacca A., Ribatti D., Ruco L. et al. Angiogenesis extent and macrophage density increase simultaneously with pathological progression in B-cell non-Hodgkin's lymphomas. Br J Cancer 1999; 79: 965-970.
Zhao W.L., Mourah S., Mounier N. et al. Vascular endothelial growth factor-A is expressed both on lymphoma cells and endothelial cells in angioimmunoblastic T-cell lymphoma and related to lymphoma progression. Lab Invest 2004; 84: 1512-1519.
Koster A., van Krieken J.H., Mackenzie M.A. et al. Increased vascularization predicts favorable outcome in follicular lymphoma. Clin Cancer Res 2005; 11: 154-161.
Aggarwal D., Srivastava G., Gupta R. et al. Angiogenesis in Non-Hodgkin's Lymphoma: An Intercategory Comparison of Microvessel Density. ISRN Hematol 2012; 2012: 943089.
Farinha P., Kyle A.H., Minchinton A.I. et al. Vascularization predicts overall survival and risk of transformation in follicular lymphoma. Haematologica 2010; 95 (12): 2157-2160.
Shayan R., Achen M.G., Stacker S.A. Lymphatic vessels in cancer metastasis: bridging the gaps. Carcinogenesis 2006; 27 (9): 1729-1738.
Banerji S., Ni J., Wang S.X. LYVE-1, a new homologue of the CD44 glycoprotein, is a lymph-specific receptor for hyaluronan. J Cell Biol 1999; 144: 789-801.
Wigle J.T., Oliver G. Prox1 function is required for the development of the murine lymphatic system. Cell 1999; 98: 769-778.
Wahlin B.E., Aggarwal M., Montes-Moreno S. A Unifying Microenvironment Model in Follicular Lymphoma: Outcome Is Predicted by Programmed Death-1-Positive, Regulatory, Cytotoxic, and Helper T Cells and Macrophages. Clin Cancer Res 2010; 16 (2): 637-650.
Mosser D.M. The many faces of macrophage activation. J Leukoc Biol 2003; 73 (2): 209-212.
Stix G. A malignant flame. Understanding chronic inflammation, which contributes to heart disease, Alzheimer's and a variety of other ailments, may be a key to unlocking the mysteries of cancer. Sci Am 2007; 297 (1): 60-67.
Jong D., Koster A., Hagenbeek A. et al. Impact of the tumor microenvironment on prognosis in follicular lymphoma is dependent on specific treatment protocols. Haematologica 2009; 94: 70-77.
Richendollar B.G., Pohlman B., Elson P., Hsi E.D. Follicular programmed death 1-positive lymphocytes in the tumor microenvironment are an independent prognostic factor in follicular lymphoma. Hum Pathol 2011; 42 (4): 552-557.
Clear A.J., Lee A.M., Calaminici M. et al. Increased angiogenic sprouting in poor prognosis FL is associated with elevated numbers of CD163-macrophages within the immediate sprouting microenvironment. Blood 2010; 115 (24): 5053-5056.
Andjelic B., Mihaljevic B., Todorovic M. et al. The number of lymphoma-associated macrophages in tumor tissue is an independent prognostic factor in patients with follicular lymphoma. Appl Immunohistochem Mol Morphol 2012; 20 (1): 41-46.
Álvaro T., Lejeune M., Camacho F.I. et al. The presence of STAT1-positive tumor-associated macrophages and their relation to outcome in patients with follicular lymphoma. Haematologica 2006; 91: 1605-1612.
Canioni D., Salles G., Mounier N. et al. High Numbers of Tumor-Associated Macrophages Have an Adverse Prognostic Value That Can Be Circumvented by Rituximab in Patients With Follicular Lymphoma Enrolled Onto the GELA-GOELAMS FL-2000 Trial. J Clin Oncol 2008 26 (3): 440-446.
Taskinen M., Karjalainen-Lindsberg M.-L., Nyman H. et al. A HighTumor-Associated Macrophage Content Predicts Favorable Outcome in Follicular Lymphoma Patients Treated with Rituximab and Cyclophosphamide-Doxorubicin-Vincristine-Prednisone. Clin Cancer Res 2007; 13: 5784-5789.
Leger-Ravet M.B., Devergne O., Peuchmaur M. In situ detection of activated cytotoxic cells in follicular lymphomas. Am J Pathol 1994; 144 (3): 492-499.
Alvaro T., Lejeune M., Salvado M.T. et al. J Clin Oncol 2007; 25 (10): 1289-1891.
Lee A.M., Clear A.J., Calaminici M. et al. Number of CD4 Cells and Location of Forkhead Box Protein P3-Positive Cells in Diagnostic Follicular Lymphoma Tissue Microarrays Correlates With Outcome. J Clin Oncol 2006; 24 (31): 5052-5059.
Laurent C., Müller S., Do C. et al. Distribution, function, and prognostic value of cytotoxic T lymphocytes in follicular lymphoma: a 3-D tissue-imaging study. Blood 2011; 118 (20): 5371-5379.
Nesterova E.S., Kravchenko S.K., Gemdzhyan E.G. i dr. Lechenie follikulyarnoi limfomy: 10-letnii opyt. Gematol i transfuziol 2012; 5: 3-8.
Pokrovskaya O.S., Mendeleeva L.P., Kaplanskaya I.B. i dr. Angiogenez v kostnom mozge bol'nykh mnozhestvennoi mielomoi na razlichnykh etapakh vysokodoznoi khimioterapii. Klin onkogematol 2010; 4: 347-353.
Lyubimova L.S., Savchenko V.G., Mendeleeva L.P. i dr. Transplantatsiya allogennogo kostnogo mozga pri khronicheskom mieloleikoze. Ter arkh 2004; 7: 18-24.
Pivnik A.V., Samoilova R.S., Novikov V.A. i dr. Differentsial'no-diagnosticheskoe i prognosticheskoe znachenie pokazatelei syvorotochnykh tsitokinov pri limfogranulematoze i limfosarkomakh. Ter arkh 2001; 4: 45-51.
Magomedova A.U., Kravchenko S.K., Kremenetskaya A.M. i dr. Modifitsirovannaya programma NHL-BFM-90 dlya lecheniya bol'nykh diffuznoi V-krupnokletochnoi limfosarkomoi. Ter arkh 2006; 10: 44-47.
Magomedova A.U., Kravchenko S.K., Kremenetskaya A.M. i dr. Effektivnost' kursa CHOP-21 v terapii diffuznoi V-krupnokletochnoi limfosarkomy. Ter arkh 2005; 7: 58-61.
Swerdlow S.H., Campo E., Harris N.L. et al. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. Lyon: IARC Press; 2008.