Vimentin immunostaining in fibroblastic reticulum cells within human reactive and neoplastic lymphoid follicles

We analyzed the distribution of fibroblastic reticulum cells (FRCs), stationary cells of lymphoid tissues, as visualized by the anti-vimentin (V9) monoclonal antibody in human reactive and neoplastic lymphoid follicles, by using immunoenzymatic and immunofluorescence methods on fixed and paraffin-embedded tissue sections from 37 lymphoid specimens with reactive disorders and 10 specimens with nodular/follicular non-Hodgkin's lymphomas (NHLs). The pattern of distribution of the vimentin-positive (VIM+) FRCs was compared with that of follicular dendritic reticulum cells (DRCs) as visualized by anti-S-100 protein antibody. Elongate VIM+ FRCs intimately attached to reticulum fibers were randomly distributed in the paracortical and interfollicular areas of lymph nodes, whereas they were recognized specifically in the mantle zones of the secondary follicles, mostly in the outer margins. Germinal centers were consistently devoid of VIM+ FRCs. Comparative analysis on serial sections as well as paired immunoperoxidase and double immunofluorescence studies demonstrated that there was a sharp difference between the patterns of intrafollicular distribution of VIM+ FRCs and S-100 protein-positive (S-100+) DRCs without juxtaposition, the FRCs being confined to the mantle zones. In the 10 nodular/follicular NHLs VIM+ FRCs could be observed in the thinned mantles of neoplastic nodules displaying a corona-like pattern that accentuated the boundaries of the nodules. The results of this study support the view that the intrafollicular distribution of VIM+ FRCs is specific for the mantle zone. The different microenvironmental organization within the follicles of VIM+ FRCs and S-100+ DRCs suggests that FRCs or at least VIM+ FRCs are stationary cells strictly related to the mantle zone microenvironment, where they may play a role in supposed sustentacular and immunologic functions similar to that of DRCs in the germinal center microenvironment.     

Significance of C4d deposition in the follicular lymphoma and MALT lymphoma and their relationship with follicular dendritic cells

We evaluated the deposition of C4d in follicular lymphomas (FL) and extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT lymphoma). Deposition of C4d was detected in 118 lymphoma tissues from patients with lymphoma and in 20 reactive hyperplasia lymphadens (RHL) using immunohistochemistical methods. FL, MALT lymphoma, and RHL were studied using double staining for CD35/C4d and Bcl-2/C4d. We studied 26 FL tissues, 19 of which showed C4d deposition. C4d deposition was detected around the follicular dendritic cells (FDCs) in the neoplastic follicles. There was no significant difference between the positive ratio of C4d and the grades of FL. We studied 12 MALT lymphoma tissues, six of which displayed C4d deposition. In these tissues, C4d deposition was detected in the peripheral region of partially colonized follicles in the form of an irregular ring, but was not found in the central region. C4d deposition was negative in completely colonized follicles. There was no C4d deposition in diffuse large B-cell lymphomas, mantle cell lymphomas, B-small lymphocytic lymphomas, T-lymphoblastic lymphomas, peripheral T-cell lymphomas, and anaplastic large cell lymphomas. C4d around the FDCs in the neoplastic follicles was a specific indicator for FL. C4d deposition in partially colonized follicles of MALT lymphoma was completely different from that in neoplastic follicles of FL, forming a key point for differential diagnosis.     

Immunohistochemical detection of follicular dendritic cells in the diagnosis of small B-cell lymphomas

Our study included 46 cases of B-chronic lymphocytic leukemia/small lymphocytic lymphoma (B-CLL/SLL), 25 mantle cell lymphomas (MCL), and 40 follicular lymphomas (FL). The lymphomas were diagnosed according to current morphological, immunohistochemical, and in some cases molecular-genetic criteria. The patterns of follicular dendritie cells (FDCs) were studied in paraffin sections using the anti CD21, anti CD23, and CNA.42 antibodies. The differences in staining patterns of FDCs among the studied lymphomas were statistically significant. The absence or small number of FDC clusters are typical of B-CLL/SLL, while FDC meshwork limited to peripheral parts of neoplastic follicles is characteristic for FL. In contrast, MCL is characterized by irregular FDC meshwork or by FDC clusters present in the centres of residual follicles. Thus, we have demonstrated that the patterns of FDCs is an independent diagnostic feature for distinguishing between B-CLL, FL, and MCL.     

Follicular dendritic cell immunohistochemical markers in angioimmunoblastic T-cell lymphoma.

Angioimmunoblastic T-cell lymphoma is characterized by a paracortical proliferation of medium to large neoplastic T cells, often with clear cytoplasm, in a background of arborizing high endothelial venules, many surrounded by follicular dendritic cells (FDCs). IHC staining may be applied to highlight these extrafollicular FDCs, traditionally using CD21, or CD23. Several alternative FDC markers have been described, including CNA.42, cystatin A/acid cysteine proteinase inhibitor (ACPI, involved in antigen presentation), and fascin (an actin binding protein). The authors stained a collection of 45 angioimmunoblastic T-cell lymphomas with CD21, CD23, CNA.42, cystatin A, and fascin for direct comparison of FDC staining characteristics in this setting. CD21 highlighted the expected dendritic network of cell processes, within residual follicles and outside of follicles, often adjacent to proliferating vessels. CD23 exhibited similar staining quality but was less sensitive than CD21. CNA.42 showed only diffuse weak labeling of FDCs. Cystatin A stained the cytoplasm of follicular dendritic cells within and outside of follicles; however, staining was often not sharply localized to dendritic cell processes, and scoring was further complicated by reactivity with other cell types in over half of the cases. Likewise, fascin stained a variety of cell types, including strong staining of interdigitating dendritic-like cells, moderate staining of endothelial cells, and only weak staining of follicular dendritic cells within and outside of follicles. Thus, CD21 remains the most reliable marker of follicular dendritic cells in angioimmunoblastic T-cell lymphoma.     

IgM expression in paraffin sections distinguishes follicular lymphoma from reactive follicular hyperplasia

The trapping of IgM-containing immune complexes (ICs) by follicular dendritic cells (FDCs) serves as an important step in promoting germinal center (GC) formation. Thus, the deposition of IgM-containing ICs on FDCs can be detected by antibodies recognizing IgM. The present investigation provides the first comprehensive report on the IgM staining pattern in follicular lymphoma (FL, n = 60), with comparisons to reactive follicular hyperplasias (RFH, n = 25), demonstrating that immunohistochemical staining for IgM in paraffin-embedded sections seems to be an additional tool for differentiating between FL and RFH. In RFH, IgM highlighted processes of FDCs, with stronger and more compact staining in light than in dark zones, with occasional very dim staining of GC B cells. In FL, IgM expression patterns were of three types. Pattern I (38 cases) stained tumor cells within neoplastic follicles, with no staining of FDCs. Pattern II (15 cases) stained neither tumor cells nor FDCs. Pattern III (7 cases) stained tumor cells with (3 cases) or without (4 cases) IgM expression; however, variable and attenuated IgM expression was observed on FDCs in each case. Interestingly, significant numbers of IgD+ mantle cells were preserved around the neoplastic follicles in these 7 cases. The data suggested that a complete or considerable loss of IgM expression in FDCs, reflecting the loss of IgM-containing ICs in FDCs, is a typical feature of FL. Increased IgM expression by GC B cells can also serve as an indicator of immunophenotypic abnormality in FL.     

The adhesion molecules, l-selectin and sialyl lewis x, relate to the formation of the follicular dendritic cell-lymphocyte cluster in the mantle zone

The follicular dendritic cell (FDC)-lymphocyte cluster is rich in the follicular light zone of the secondary lymphoid follicles (LFs). Although, the mantle zone (MZ) also has FDC-lymphocyte cluster, it has not known about what kind of adhesion molecules relates to cluster formation. In the present study, we investigated whether the adhesion molecules, L-selectin (CD62L) and sialyl Lewis x (CD15s) can mediate the formation of the cluster in human tonsillar LFs. The MZ only expressed both the adhesion molecules in the secondary LF. Isolated FDC-lymphocyte clusters were composed of CD62L(+) lymphocytes and CD15s(+) FDCs. Stamper-Woodruff binding assay revealed that the binding of IgD(+) lymphocytes was significantly inhibited by pretreatment with anti-CD62L antibody or with anti-CD15s antibody. These results indicate that CD62L on MZ lymphocytes and CD15s on FDCs may play a role of the cluster formation, unlike the clusters in the other parts of LFs.     

Protection of germinal centres from complement attack: Decay-accelerating factor (DAF) is a constitutive protein on follicular dendritic cells. A study in reactive and neoplastic follicles

The development of B-cell memory is linked to the presence of germinal centres. This process is dependent on the presence of antigen, usually in the form of immune complexes with antibody, on the surface of the follicular dendritic cells (FDCs) that form a network in the germinal centre. The presence of immune complexes poses a constant danger of activating complement. Decay accelerating factor (DAF, CD55) and the membrane attack complex (MAC) inhibitor (CD59) are two cell proteins whose sole function is to protect cells from the action of complement, the former affecting the earlier components of the complement cascade, and the latter the terminal ones; both are bound to the cell surface via a glycosylphosphatidylinositol link. DAF but not CD59 could be demonstrated on FDCs. DAF is also present on the FDCs in follicular lymphomas despite the absence of complement (C3) in neoplastic follicles. This indicates that DAF is constitutive to FDCs but does not preclude the possibility that its expression is increased when immune complexes are deposited.     

The Leukocyte Semaphorin CD100 Is Expressed in Most T-Cell, but Few B-Cell, Non-Hodgkin’s Lymphomas

The 150-kd transmembrane protein CD100 is the first semaphorin protein shown to be expressed in lymphoid tissue. CD100 is present in the interfollicular T cell zones and is also expressed by B cells in the germinal centers of secondary lymphoid follicles, but not in the mantle zones. The CD100 molecule was recently cloned, and CD100 transfectants were shown to induce homotypic aggregation of human B cells and improve their viability in vitro, suggesting that CD100 may play a role in lymphocyte aggregation and germinal center formation. We studied the expression of CD100 in 138 clinical cases representing a range of lymphoproliferative disorders, to determine whether this molecule is expressed in these neoplastic processes. In general, we found CD100 expression to be common in peripheral T-cell non-Hodgkin’s lymphomas but rare in B-cell non-Hodgkin’s lymphomas. CD100 expression was not detectable in low-grade B-cell non-Hodgkin’s lymphomas, including cases of small lymphocytic lymphoma (18 cases), marginal zone lymphoma (10 cases), and mantle cell lymphoma (10 cases), as might be expected for these neoplasms that are not of follicular center cell origin. Surprisingly, we found that the vast majority of follicular lymphomas (37 of 40 cases) as well as diffuse large-cell lymphomas of B-cell type (35 cases) did not express CD100. The neoplastic cells in 3 of 11 cases of predominantly large-cell-type follicular lymphoma did express CD100. In contrast, all five cases of high-grade, small non-cleaved (Burkitt-like) B-cell lymphoma were immunoreactive for CD100 expression, as were 18 of 20 cases (90%) of malignant T cell neoplasms. Northern blot analysis of CD100 expression correlated with immunohistochemical findings. Absence of expression of CD100 by neoplastic follicular center B cells is a common feature in follicular lymphomas, but expression of CD100 by T cells is maintained in T-cell lymphoproliferative disorders.     

Disappearance of CD21-positive follicular dendritic cells preceding the transformation of follicular lymphoma: immunohistological study of the transformation using CD21, p53, Ki-67, and P-glycoprotein

Some follicular lymphomas histologically transform into diffuse aggressive lymphomas, the prognosis of which is poor. There are, however, no reliable histological criteria for predicting which cases will later undergo such transformation. In low-grade B-cell lymphomas, follicular dendritic cells form dense mesh-like networks that contain accumulating neoplastic B-cells. These are rare in high-grade lymphomas. We immunohistochemically analyzed CD21-positive follicular dendritic cells in 32 follicular lymphomas, including 3 transformed lymphomas, in addition to immunohistological study using P-glycoprotein, p53, and Ki-67. We found that the mesh-like networks in follicles are more clearly defined in low-grade lymphomas than in high-grade lymphomas (p = 0.015). Neoplastic follicles in 2 transformed lymphomas lost the networks of follicular dendritic cells before transformation despite the existence of morphologically clear follicles. This differed from the non-transformed cases of the same cytological grades. Prognosis was statistically better for patients with low-grade tumor than for those with high-grade tumor (p = 0.026), and there was a trend toward poorer survival among CD21-negative cases (p = 0.186). P-glycoprotein, p53, and Ki-67 expressions did not provide sufficient information to predict the transformation of follicular lymphoma. The presence of CD21-positive follicular dendritic cells in neoplastic follicles might help predict the potential of follicular lymphoma to transform to diffuse large B-cell lymphoma.     

Relevance of Ki-67 expression in the classification of non-Hodgkin''s lymphomas: a morphometric and double-immunostaining study

The proliferation of reactive and neoplastic cells was retrospectively assessed in 92 cases of non-Hodgkin's lymphoma by morphometry using a double-immunoenzymatic technique including surface markers and the monoclonal antibody Ki-67. The findings were compared with the histological diagnosis. The overall Ki-67 positivity is not always a good measure of the corresponding corrected values and therefore we recommend that a correction should be made for the total number of complementary lymphocytes in the tumour. Taking the macrophages and the Ki-67 positivity of the reactive cells into account does not generally add any information. There was no difference in reactive cell content between follicular (counted within follicles) and diffuse lymphomas within the tumour areas. The value of the group mean for low-grade follicular (nodular) lymphomas was significantly higher than that of diffuse low-grade lymphomas, but not significantly different from that of intermediate-grade lymphomas. High-grade lymphomas exhibited significantly greater Ki-67 positivity than those of intermediate grade. In 76% of the cases there was significant agreement between malignancy grade (low/intermediate malignant versus high malignant) at 45% corrected Ki-67 counts.     

Monoclonal antibody 4C7 recognizes an endothelial basement membrane component that is selectively expressed in capillaries of lymphoid follicles

In order to define compartment-related structures within the extracellular matrix of human lymphoid organs, monoclonal antibodies (MAbs) were generated by immunizing mice with stromal fragments of human tonsils. One MAb (4C7) was selected which recognized an endothelial basal membrane component that is selectively expressed in capillaries of lymphoid follicles. The epitope was also present in follicles within chronically inflamed synovial membrane and in a hyperplastic thymus of a patient with myasthenia gravis. B-cell non-Hodgkin's lymphomas with a follicular growth pattern expressed the antigen in neoplastic follicles, whereas diffuse growing lymphomas lacked the antigen. The restricted distribution pattern suggests involvement of the 4C7-defined antigen in the organization of the follicular compartment within human lymphoid tissue.     

Distribution of T-cell subsets in follicular and diffuse lymphomas of B-cell type.

The authors examined the number and distribution of cells reacting with monoclonal antibodies to T-cell subsets in frozen tissue sections of B-cell lymphomas (30 follicular and 17 diffuse lymphomas). In five diffuse lymphomas (two lymphocytic, three small cleaved cell) the neoplastic B-lymphocytes reacted with the monoclonal antibody anti-T1. In all other cases, the monoclonal antibodies to T-cell subsets reacted only with small lymphocytes concentrated between the follicles of follicular lymphomas and distributed randomly in diffuse lymphomas. The distribution of T cells and the T4+/T8+ ratio in follicular small cleaved and mixed lymphomas was similar, although not identical, to that seen in hyperplastic lymphoid follicles. Fewer T cells and a decrease in the T4+/T8+ ratio were seen in follicular large cell lymphoma and in diffuse large cell lymphomas. The number and distribution of T cells in follicular lymphomas is consistent with the hypothesis that there is a functional interaction between neoplastic B cells and benign T cells. No tumors were found in which the neoplastic B cells reacted with anti-T3, anti-T4, or anti-T8.     

Nerve growth factor receptor expression on dendritic reticulum cells in follicular lymphoid proliferations.

Using an antibody to the nerve growth factor receptor (NGFR), we examined dendritic reticulum cells (DRCs) immunohistochemically in 62 formalin-fixed, paraffin-embedded lymph nodes from patients with reactive follicular hyperplasia or with various types of lymphoma. A dendritic staining pattern within germinal centers was present in 25 of 26 routinely processed lymph nodes with reactive follicular hyperplasia. In contrast, dendritic staining with anti-NGFR was present within neoplastic follicles in only three of 28 follicular lymphomas. Staining of benign, residual germinal centers with anti-NGFR was present in mantle zone lymphoma and Hodgkin's disease. These findings suggest a possible role for the NGFR in the maturation and/or activation of normal DRCs. The loss of NGFR expression in most follicular lymphomas indicates that DRCs are altered as part of the neoplastic process. The possibility that DRCs may play a role in the pathogenesis of follicular lymphoma is suggested.     

CD40 ligand is constitutively expressed in a subset of T cell lymphomas and on the microenvironmental reactive T cells of follicular lymphomas and Hodgkin's disease.

Although CD40 has been extensively studied in B- and T-cell non-Hodgkin's lymphomas (NHLs)/leukemias, and more recently in Hodgkin's disease (HD), little is known about the expression of its ligand (CD40L) in lymphoproliferative disorders other than T-cell NHLs/leukemias. A series of 121 lymphoma/leukemia samples, including 35 cases of HD, 34 T-cell and 39 B-cells NHLs, 2 cases of adult T-cell leukemia/lymphoma, and 11 cases of T-cell acute lymphoblastic leukemia, were evaluated for CD40L expression by immunostaining of frozen tissue sections and flow cytometry with the anti-CD40L monoclonal antibody M90. CD40L was constitutively expressed by neoplastic cells in 15 of 36 (42%) T-cell NHLs/adult T-cell leukemia/lymphomas, almost invariably those displaying the CD4+/CD8- phenotype, whereas no CD40L-expressing tumor cells could be found in B-cell NHL and HD. Among T-cell acute lymphoblastic leukemias, CD40L was detected only on 2 cases displaying a stem-cell-like phenotype. In follicular B-cell lymphomas a large number of CD40L-expressing CD3+/CD4+ T lymphocytes were found admixed with tumor cells within the neoplastic follicles and in their surrounding areas. In the nonfollicular B-cell lymphomas, CD40L-positive CD3+/CD4+ T lymphocytes were few or absent. In all HD subtypes other than the nodular lymphocytic predominance, CD40L-expressing CD3+/CD4+ T lymphocytes were numerous in the HD-involved areas and were mainly located in close proximity to the Reed-Sternberg cells. Our data indicate that in human lymphomas CD40L is preferentially expressed by a restricted subset of T-cell lymphomas, mostly with CD4 immunophenotype. Finally, we have provided morphological evidence that CD40L may play an important role in the cell contact-dependent interaction of tumor B-cells (CD40+) within the neoplastic follicles or Reed-Sternberg cells (CD40+) in HD-involved areas and the microenvironmental CD3+/CD4+/CD40L+ T lymphocytes.     

B- and T-cell subsets in follicular centroblastic/centrocytic (cleaved follicular center cell) lymphoma: an immunohistologic analysis of 26 lymph nodes and three spleens

Follicular centroblastic/centrocytic (CB/cc/F), or cleaved follicular center cell, lymphomas are known to contain admixtures of B cells and, often, numerous T cells. To analyze the presence and distribution of B and T cells and their subsets in CB/cc/F lymphomas, 26 lymph nodes and three spleens (from 24 patients) were studied with a panel of monoclonal antibodies and peanut lectin by the avidin-biotin immunoperoxidase technique on frozen sections. Immunoglobulin studies revealed monoclonal neoplastic follicles in most cases, although cells of the nondominant light chain were occasionally present. Rarely, the follicles showed both kappa and lambda light chains or were immunoglobulin-negative. Although monoclonal mantles were observed in only one case, more than half of the nodes demonstrated monoclonal interfollicular infiltrates. In most cases the phenotype of the follicular cells was similar to that of the predominant cells in the normal follicles. Interfollicular lymphomatous cells had variable phenotypes that, unlike the normal situation, sometimes resembled those in the follicles. Phenotypic variation was present within single neoplastic clones and sometimes suggested more mantle-like differentiation. Phenotypic changes were also observed in repeat biopsies. T cells, usually predominantly of the T-helper phenotype, were present in variable numbers in all cases. Although present in the neoplastic follicles, they were usually more common in the interfollicular areas. Six nodes and two spleens had definite rims composed predominantly of T cells around neoplastic follicles. Apparently "activated" (Tac-positive) T cells were often present and showed accentuation around the follicles in some cases. Thus, CB/cc/F lymphoma is a malignant lymphoma that closely resembles follicular hyperplasia but that also has distinctive features.     

Identification of CXCL13 as a new marker for follicular dendritic cell sarcoma

The homeostatic chemokine CXCL13 is preferentially produced in B-follicles and is crucial in the lymphoid organ development by attracting B-lymphocytes that express its selective receptor CXCR5. Follicular dendritic cells (FDCs) have been identified as the main cellular source of this chemokine in lymphoid organs. Recently, genome-wide approaches have suggested follicular CD4 T-helper cells (T(H)F) as additional CXCL13 producers in the germinal centre and the neoplastic counterpart of T(H)F (CD4+ tumour T-cells in angioimmunoblastic T-cell lymphoma) retains the capability of producing this chemokine. In contrast, no data are available on CXCL13 expression on FDC sarcoma (FDC-S) cells. By using multiple approaches, we investigated the expression of CXCL13 at mRNA and protein level in reactive and neoplastic FDCs. In reactive lymph nodes and tonsils, CXCL13 protein is mainly expressed by a subset of FDCs in B-cell follicles. CXCL13 is maintained during FDC transformation, since both dysplastic FDCs from 13 cases of Castleman's disease and neoplastic FDCs from ten cases of FDC-S strongly and diffusely express this chemokine. This observation was confirmed at mRNA level by using RT-PCR and in situ hybridization. Of note, no CXCL13 reactivity was observed in a cohort of epithelial and mesenchymal neoplasms potentially mimicking FDC-S. FDC-S are commonly associated with a dense intratumoural inflammatory infiltrate and immunohistochemistry showed that these lymphocytes express the CXCL13 receptor CXCR5 and are mainly of mantle zone B-cell derivation (IgD+ and TCL1+). In conclusion, this study demonstrates that CXCL13 is produced by dysplastic and neoplastic FDCs and can be instrumental in recruiting intratumoural CXCR5+ lymphocytes. In addition to the potential biological relevance of this expression, the use of reagents directed against CXCL13 can be useful to properly identify the origin of spindle cell and epithelioid neoplasms.     

Calmodulin immunoreactivity in the chicken pineal gland: Comparison with calbindin-D28k,calretinin, and S100

Calmodulin distribution in the chicken pineal organ was investigated by immunohistochemistry. Calmodulin immunoreactivity was detected in ependymocytes in the follicular zone and in interstitial cells in the parafollicular zone. No calmodulin immunoreactivity was detected in pinealocytes. Lack of calmodulin immunoreactivity in pinealocytes raises questions about its proposed function in melatonin synthesis as suggested by pharmacological studies using calmodulin antagonists. The calmodulin distribution was comparable to that of S100, a glial cell marker. Two other markers, calbindin-D28k and calretinin, which in neuroanatomical studies give excellent cytoarchitectonic staining, in the chick pineal permitted the detection of two subclasses of pinealocytes. One was darkly stained by calbindin-D28k and rare. The other was very abundant and calretinin positive. In the parafollicular zone, calbindin-D28k and/or calretinin antibodies allowed us to visualize cells presenting a neuron-like morphology. Calretinin immunoreactivity was detected in nearly all pinealocytes in which hydroxy-indol-O-methyl transferase was also located. Comparison between the lack of calmodulin and the presence of calretinin, belonging to the same calcium-binding protein family, in chick pinealocytes raises the hypothesis about a possible role of calretinin in melatonin synthesis. © 1994 Wiley-Liss, Inc.     

DRC antigen expression in B-cell lymphomas

DRC (dendritic reticulum cell) antigen expression was studied in 38 cases of B-cell lymphomas including follicular lymphoma. The results of this study showed DRC-1 to be expressed in 1/3 of small lymphocytic; 3/3 of mantle zone lymphoma (MZL); 10/10 of follicular, small cleaved; 6/7 of follicular, mixed; 1/2 of follicular, large cell lymphomas. However, DRC-1 was not expressed in any of diffuse, small cleaved (0/6) and diffuse, large cell (0/6). Although S-100 protein was positive in the majority of these DRC-1-positive cases on the paraffin embedded specimens, positive nodules were less intense and smaller in number compared with those of DRC-1 on frozen tissue specimens. These results suggest that in case of small lymphocytic lymphoma found to be positive for DRC-1, the networks of the DRCs are expressed in the pseudofollicular proliferation centers. This study also suggests that the networks of the DRCs newly appear accompanying the neoplastic growth rather than originating from residual germinal centers, and that neoplastic small cleaved cells play a major role in inducing the DRCs in the positive cases of follicular lymphoma, MZL, and small lymphocytic lymphoma.     

Cellular composition of follicles of follicle centre cell lymphomas in relation to germinal centres of reactive lymph nodes. A morphometrical electromicroscopical study

The cell spectrum of neoplastic and benign reactive germinal centres was determined ultrastructurally. The degree in which the cell composition found in reactive germinal centres is maintained in analogous structures of follicular lymphomas was investigated by pattern recognition methods and discriminant analysis based on the frequencies of the various lymphoid and non-lymphoid cell types. The follicular lymphomas included lymphomas with predominantly centrocytes (FCCL, Cb-cc) and lymphomas with predominantly centroblasts (FCCL, Cb). Pattern analysis of FCCL Cb, FCCL Cb-cc and reactive germinal centres indicates that FCCL Cb follicles resemble reactive germinal centres in more aspects than follicles of FCCL Cb-cc. Clear statistical differences were encountered between the frequencies of the lymphoid cell types and of the follicular dendritic and histiocytic reticulum cells in follicles of FCCL Cb-cc and FCCL Cb and reactive germinal centres. Application of a discriminant analysis using a combination of the frequency of centrocytes and follicular dendritic cells demonstrated that both types of neoplastic follicles and reactive germinal centres were correctly classified on the basis of their cell spectrum. For the three groups the most potent discriminator was the centrocyte, whereas the small and large centroblast were of less value. For discrimination between Cb-cc follicles and reactive germinal centres again the centrocyte was the most potent discriminator. Discrimination between FCCL Cb follicles and reactive germinal centres of FCCL Cb-cc follicles can be easily achieved using the frequencies of small centroblasts or centrocytes on their own. These findings indicate that (1) follicles of both FCCL Cb and FCCl Cb-cc differ greatly in the cellular composition and not only with respect to their content of centroblasts but also in their content of follicular dendritic cells and (2) they may be considered as neoplasms representing different developmental phases of germinal centres.     

Cluster differentiation antigen expression, proliferative activity and clinical stage in centroblastic centrocytic lymphomas

Using a panel of B-cell antibodies recognizing clusters of leucocyte differentiation antigens, immunostaining patterns of eight reactive lymph nodes and 28 centroblastic/centrocytic and centrocytic lymphomas have been studied. Centroblastic/centrocytic and centrocytic lymphomas retained many of the B-cell differentiation antigens and neoplastic follicles partially recapitulated the staining patterns observed in reactive follicles. Centrocytic lymphomas usually expressed a heavy chain mantle zone-like phenotype. Nearly one-half of follicular lymphomas showed extension of neoplastic cells into interfollicular areas as evidenced by positivity for CD10 (common acute lymphoblastic leukaemia) and/or CD9 (immature B-cell) and CD23 (B-blast cell) antigens. Cases showing interfollicular involvement also manifested considerable phenotypic heterogeneity. Light chain restriction could not be used to determine interfollicular involvement because of the presence of many non-neoplastic cells. Most follicular lymphomas retained a polyclonal mantle around at least some neoplastic follicles and in no case was a monoclonal mantle seen. Most lymphomas (16/21) were diploid when examined by flow cytometry. Diploid tumours exhibiting interfollicular lymphomatous involvement had high proliferation (S + G2) fractions and these lymph nodes were usually derived from patients with widespread disease. Tumours containing a high percentage of cells in the G0/G1 phases displayed fewer B-cell differentiation antigens than tumours with low G0/G1 fractions.