Follicular Dendritic Reticulum Cells Research Articles

Expression of ICAM-1, VCAM-1 and ELAM-1 in angiofollicular lymph node hyperplasia (Castleman's disease): evidence for dysplasia of follicular dendritic reticulum cells

The inducible adhesion molecules mediate important functions in the lymphoid tissues. We have investigated the expression of intercellular adhesion molecule 1 (ICAM-1), endothelial leucocyte adhesion molecule 1 (ELAM-1), vascular cell adhesion molecule 1 (VCAM-1), and platelet endothelial cell adhesion molecule (PECAM/CD31), using immunocytochemistry on cryostat sections of five lymph nodes from patients with Castleman's disease of the hyaline-vascular type. All five cases were characterized by marked hyperplasia of follicular dendritic reticulum cells, which were extensively present even in the mantle zone. Hyperplastic follicular dendritic reticulum cells showed marked expression of VCAM-1, and weak expression of ICAM-1. In two cases, several dysplastic giant cells with aberrant, polyploid nuclei showed aberrant expression of ELAM-1, an endothelium-restricted molecule. Dysplastic giant cells were positive with DRC-1 (an antibody to dendritic reticulum cells), VCAM-1 and occasionally ICAM-1, were negative for the endothelial cell markers factor VIII-related antigen and CD31 and were non-proliferating (Kl-67-). Cells positive for ICAM-1 or VCAM-1 were rare in the interfollicular areas. In all cases vascular hyperplasia was prominent, but endothelial cells were poorly activated in terms of expression of inducible adhesion molecules and of HLA-DR antigens. The possibility that dysplastic follicular dendritic reticulum cells have a pathogenetic role in Castleman's disease is discussed.

Ki-M6 Immunostaining in Routinely Processed Sections of Reactive and Neoplastic Human Lymphoid Tissue

A monoclonal antibody, termed Ki-M6 (CD68), which shows a restricted reactivity to cells of the monocyte/macrophage system, has been evaluated primarily with the use of cryostat sections. In this study the authors could assess that the Ki-M6 antibody recognizes a fixation-resistant epitope in most human macrophages. The Ki-M6 immunoreactivity with monocyte/macrophage-related cells was established by testing on routinely processed samples of reactive and neoplastic lymphoid tissues; it was compared with the staining for vimentin (V9) and S-100 protein antibodies, with visualization of the stationary elements of lymphoid tissues, with the aim of establishing its value in the study of the nonlymphoid microenvironment. The Ki-M6 antibody reactivity could be achieved with Bouin-fixed, paraffin-embedded tissue sections, without any proteolytic treatment, with the use of the avidin-biotin complex (ABC) method, especially after overnight incubation time at 4 degrees C. Some reduction in antigenic reactivity was observed in B5- or formaldehyde-fixed samples. The antibody reacted with macrophages of all different lymph node compartments; a broad reactivity against cells of macrophage lineage, including multinucleated giant cells, was observed in epithelioid granulomas. Ki-M6-positive cells other than classic macrophages were the so-called "plasmacytoid T-cells" and cells displaying elongated cytoplasms with fibroblastic-like features. Granulocytes, follicular dendritic reticulum cells, and interdigitating reticulum cells did not reveal any reactivity with Ki-M6 antibody. In malignant lesions, neoplastic cells of follicular and diffuse B- and T-cell lymphomas, including large cell non-Hodgkin's lymphomas, and Reed-Sternberg cells of Hodgkin's disease were negative in all cases studied. This study shows that Ki-M6 seems to be another anti-macrophage-specific antibody that reacts, in routinely processed tissue sections, with tissue macrophages but not with accessory cells. Thus, it may be a valuable addition to vimentin and S-100 protein antibodies for investigation of the microenvironmental organization of lymphoid tissues both in normal and neoplastic conditions.

Pathological and immunohistochemical studies of follicular lymphoma in two calves

Pathological and immunohistochemical studies were performed on two cases of bovine follicular lymphoma with severe leukaemia and antibodies to bovine leukaemia virus. Histologically, numerous neoplastic follicular structures were observed in the lymph nodes and spleen of both cases and in the tonsils and Peyer's patches of one case. The neoplastic cells infiltrated diffusely into the liver, lungs and heart in both cases and into the tonsils, kidneys, abomasum and bone marrow in one case. Intracytoplasmic immunoglobulin was negative in the neoplastic cells. Follicular dendritic reticulum cells were decreased in number in the neoplastic follicles compared with those in normal lymphoid follicles. Follicular lymphoma is rare in cattle. Histological features of the present cases are distinguishable from the adult form of bovine lymphoma and the calf form of lymphoma.

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.

Dendritic reticulum cell-related immunostaining for laminin in follicular and diffuse B-cell lymphomas

We performed a comparative immunohistocytochemical study of the distribution patterns of laminin and follicular dendritic reticulum cells (DRCs) within their follicular microenvironment in both nodular or diffuse B-cell non Hodgkin's lymphomas (NHLs). Twenty nine cases of immunophenotypically diagnosed B-cell NHLs (19 of follicular center cell origin-FCCL- and 10 of the diffuse well differentiated lymphocytic type-WDLL-) and five reactive lymph nodes with follicular hyperplasia were analyzed by immunoperoxidase and immunofluorescence techniques. Serial frozen sections and cytospin preparations were tested either with single antibodies anti laminin and DRC-1, or paired reagents in double labeling immunofluorescence. Our results indicated consistently that within both the reactive germinal centers and the neoplastic nodules of FCCL laminin immunostaining visualized a punctate-granular pattern apart from the linear vascular basement membrane positivity. Double immunofluorescence assay demonstrated that there was a close parallelism between this laminin staining pattern and DRC-1 distribution showing a well developed DRCs meshwork; in the diffuse tumour areas of both FCCL and WDLL, laminin immunoreactivity was found only in those cases in which nests of DRCs were observed. Double immunofluorescence studies performed on cytospin preparations demonstrated that the groups of cells containing DRC-1 positive cells, contained a positivity for laminin, although within the cell the staining for DRC-1 was intense and diffuse, while that for laminin was granular and more sparse. Our results suggested that these laminin and DRC-1 positive reactive sites may be present on the same cells. Since the reduction in number or loss of both DRCs and their related immunostaining for laminin within the microenvironment was consistently associated with a loss of nodularity by lymphoma cells, whereas nodularity in reactive and neoplastic conditions was associated with a rich DRCs meshwork and the related laminin immunostaining, a trapping function of DRCs exercised in the presence of laminin should be considered.

Immunohistopathogenesis of persistent generalized lymphadenopathy in HIV-positive patients

Persistent generalized lymphadenopathy (PGL) is a reactive lymphadenitis affecting HIV-positive patients; furthermore, PGL is often a prodrome of AIDS-related complex and AIDS. In the present review the authors describe the histology and the immunohistochemistry of lymph nodes of patients affected by PGL. Histologic alterations of lymph nodes with PGL are classified according to three main types: follicular hyperplasia without or with follicular fragmentation, follicular involution and follicular depletion. Immunohistology demonstrates a peculiar infiltration of CD3+/CD4+ and CD3+/CD8+ lymphocytes in germinal centers; CD3+/CD8+ are often grouped in small clusters centered by a newly formed small blood vessel. Accessory follicular dendritic reticulum cells (FDRCs) of germinal centers are characterized by a positive staining for p24 and p19 HIV major core antigens. In germinal centers, FDRCs undergo progressive lysis in follicular involution and in follicular depletion. Other viral antigens, such as EBV, are infrequently seen in lymph nodes from HIV-positive patients. Paracortical areas of lymph nodes are often characterized by prominent postcapillary venule proliferations and by hyperplasia of the endothelial cells which are HLA-DR positive, often p19 and p24 positive, and occasionally express HIV genome. In conclusion, in PGL the histologic changes correlate well with the immunohistologic features; accordingly, PGL might be considered the result of abnormal immune reactions to several stimuli still incompletely known.

Basement membrane components in lymphoid follicles: Immunohistochemical demonstration and retationship to the follicular dendritic cell network

We analyzed the distribution of two basement membrane components, laminin and type IV collagen, in human reactive and neoplastic lymphoid follicles by using immunoenzymatic and immunofluorescence methods on serial frozen sections from 11 reactive lymph nodes, three palatine tonsils, and 11 immunophenotypically defined B cell non-Hodgkin's lymphomas. The patterns of distribution of these two antigens were compared with that of follicular dendritic reticulum cells (DRCs) as visualized by anti-DRC-1 and anti-desmoplakin 1 and 2 antibodies. Immunostaining for laminin and type IV collagen produced overlapping results. Germinal centers contained some vascular and fiber-like linear staining, but the most striking feature was the presence of a large amount of punctate-granular staining. This staining was present throughout the entire area of most of the germinal centers, although often more densely in the central areas. The stronger punctate-granular staining for laminin, type IV collagen, and desmoplakin 1 and 2 corresponded to the more densely stained areas for DRC-1. Double immunofluorescence assay demonstrated that there was a close similarity between the punctate-granular staining pattern of laminin and the dendritic network of DRC-1—positive cells. In the nine B cell lymphomas of follicular center origin in which DRCs were present, although distributed in different patterns, immunostaining for laminin and type IV collagen could be observed only in those areas showing immunoreactivity for DRC-1. The results of this study suggest that the punctate-granular staining pattern for laminin and type IV collagen is specific for the follicle. This finding may be strictly related to the follicular microenvironment because the pattern of distribution displayed by laminin and type IV collagen seems to be spatially related to the staining pattern of DRCs, as visualized by the anti-DRC-1 and desmoplakins antibodies, both in reactive and neoplastic conditions.

KP1: a new monoclonal antibody that detects a monocyte/macrophage associated antigen in routinely processed tissue sections.

A new monoclonal antibody, KP1, raised against a lysosomal fraction of human lung macrophages, recognises a fixation-resistant epitope in a wide variety of tissue macrophages (such as Kupffer cells germinal centre, splenic, and lamina propria macrophages), and in granulocyte precursors. Its broad reactivity with cells of the mononuclear phagocytic lineage was established by testing on routinely processed samples of normal and reactive lymphoid tissues. Interdigitating reticulum cells were unstained or showed limited cytoplasmic staining while Langerhans' cells and follicular dendritic reticulum cells were unreactive. KP1 recognises a molecule of about 110 kilodaltons in macrophage-rich human tissue when tested by either immunoprecipitation or Western blotting (although the latter procedure also shows two additional components with molecular weights of 70 and 40 kilodaltons). KP1 should be of considerable value for studying disorders of the monocyte/macrophage system, including both reactive and neoplastic states (such as true histiocytic proliferations).

Characterization of Extracellular Matrix in Non-Hodgkin's Lymphomas

Basement membrane proteins laminin, type IV collagen and aminoterminal propeptide of type III collagen (type III pN-collagen) are important components of extracellular matrix of normal lymph nodes; they are present in reticular fibres and in the walls of blood vessels and sinuses. We have studied the distribution of these proteins in 45 cases of non-Hodgkin's lymphomas representing different types of Kiel classification. Light microscopic immunohistochemistry was used on sections from formalin fixed, paraffin embedded biopsy specimens. Reticular cells were identified by using antibodies against S-100 protein, a marker for dendritic and interdigitating reticulum cells, and against cystatin A for the demonstration of follicular dendritic reticulum cells. The findings of the immunohistochemical stainings were generally similar in the different lymphoma types. Thus the method for the demonstration of extracellular matrix proteins could not give obvious aid for lymphoma classification. BM stainings proved to be efficient in the demonstration of sinuses. Marginal and medullary sinuses of the nodal cases were either totally or partly spared and open also in the cases in which there was extracapsular growth. The fibre content was variable, but generally the distribution of all the three extracellular matrix proteins was comparable with each other and mainly corresponded with the pattern seen in conventional Gomori's reticulin stain. This also holds for the tumor tissue infiltrated through the capsule and for extranodal cases. Areas of sclerosis of some cases contained only type III pN-collagen. Reticular fibres were at places seen to be closely associated with cells of spindle or dendritic shape. A few reticulum cells could be found to show cytoplasmic staining for laminin, type IV collagen and type III pN collagen. This finding suggests that the cells are synthetizing these proteins. The amount of cells staining for cystatin A or S-100 protein did not generally correlate with the amount of fibres, and most of the stained cells were not closely associated with fibres. The results show that in non-Hodgkin's lymphomas basement membrane proteins and type III pN-collagen are important components of reticular fibres. Results also indirectly support the main role of fibroblastic reticulum cells in the production of the fibres.

Heterogeneous immunostaining patterns of follicular dendritic reticulum cells in human lymphoid tissue with selected antibodies reactive with different cell lineages.

A comparative immunohistologic study of the cell density and distribution pattern of follicular dendritic reticulum cells (DRCs) within their follicular microenvironments (germinal centers and mantle zones) was performed by immunoperoxidase technique with a selected panel of antibodies either operationally specific for DRCs (DRC-1) or reported as having additional immunoreactivity with DRCs (antibodies to B- and T-cells, leukocytes, monocytes/macrophages, desmosomal components, and S-100 protein). Twenty-five biopsy specimens, including reactive lymph nodes and tonsils as well as normal spleen tissue, were analyzed. Serial frozen sections were tested either with single antibodies or paired monoclonal reagents in double-labeling procedures. Our results consistently indicated that DRCs positive for Leu M3 and BA-2 antibodies were confined to the central portion of germinal centers, whereas DRCs immunoreactive for S-100 protein and desmoplakin 1 and 2 were localized mostly in the central and pericentral portion of germinal centers. All the DRCs extending from the central portion of germinal centers to the mantle zones were labeled with DRC-1 and, unexpectedly, with OKB7 antibodies. Other immunostainings, such as those for HLA-DR antigens, common leukocyte antigen, and Leu 3a, were not contributory in defining topographic differences of DRCs within the follicle. The consistent heterogeneity of the labeling patterns appears to suggest a possible in situ immunophenotypic grouping of DRCs, and the concept of their possible heterogeneity appears to be corroborated.

Development and functional anatomy of the human tonsilla palatina.

The tonsilla palatina, belonging to the gut-associated lymphoepithelial organs (GALT), shows close morphological and functional correlation between the lymphatic tissue of mesenchymal origin and the endodermal epithelium of the second pharyngeal pouch. During the 4th fetal month epithelial crypts grow down into the connective tissue and are infiltrated by non-epithelial cells. In the underlying lymphoid tissue the first primary follicles with precursors of follicular dendritic reticulum cells can be localized already in the 14th fetal week post conceptionem (p.c.). The parafollicular areas develop into T-cell regions. Interdigitating cells in these regions as well as within the crypt epithelium express the HLA-DR antigen. They were frequently found in close contact with T-helper cells. This fact is discussed as an indication of the tonsillar crypt epithelium partly serving as T-cell region. But also B-cells and macrophages invade the crypt epithelium in great numbers with a distinctly different distribution. Natural killer (NK) cells (Leu-7-positive) are localized within the crypt epithelium. Small Leu-7-positive lymphocytes (probably precursors) have been found to be numerous in the germinal centers. The interaction of specific stationary cells in the T-cell and B-cell regions is obviously an important factor for the maturation of different subpopulations of immunocompetent lymphoid cells.

Heterogeneous in situ immunophenotyping of follicular dendritic reticulum cells in malignant lymphomas of B-cell origin.

The phenotype of follicular dendritic reticulum cells (DRC) was analyzed with monoclonal antibodies (DRC-1, OKB7, BA-2, Leu-M3, and antidesmoplakin 1 and 2) in 28 frozen biopsy specimens of both morphologically and phenotypically analyzed B-cell lymphomas and 21 normal or reactive controls. The former included 15 follicular center cell lymphomas (FCCL), four intermediately differentiated lymphocytic lymphomas (ILL), four mantle zone lymphomas (MZL), and five well-differentiated lymphocytic lymphomas (WDLL). In controls, DRC-1+ and OKB7+ DRC were localized in both follicular centers (FC) and mantle zones (MZ), but BA-2+ and Leu-M3+ DRC were confined to FC only. FCCL were usually accompanied by DRC-1+, OKB7+, and BA-2+ DRC, and either lost or maintained positively with Leu-M3 from case to case. By contrast, MZL consistently lacked BA-2+ and Leu-M3+ DRC, and was associated with DRC-1+ and OKB7+ DRC only. Desmoplakin-positive DRC occurred in variable proportions in both FCCL and MZL. As opposed to FCCL and MZL, all WDLL and all but one of the ILL (associated only with DRC-1+, OKB7+, and desmoplakin+ DRC) did not show any DRC as identifiable with the antibody panel used. Remarkably, the difference in the distribution of BA-2+ and Leu-M3+ DRC in the normal FC and MZ appears to be maintained in their neoplastic counterparts (FCCL and MZL) also. Such a difference represents an example of the possible interactions between lymphoma cells of different phenotype and their microenvironment, as portrayed by phenotypically heterogeneous DRC.

Microenvironment of thymic myoid cells in myasthenia gravis.

The microenvironment of myoid cells (MyCs) was studied in myasthenia gravis (MG) thymitis with lymphoid follicular hyperplasia (LFH) (nine cases) and with diffuse B cell infiltration (one case), and compared with findings in the thymuses of non-myasthenic control subjects (ten cases). Double immunostaining was used to demonstrate MyCs labelled by anti-desmin together with other thymic components such as keratin-positive epithelial cells, Ki-M 1-positive interdigitating reticulum cells (IDCs), Ki-M 4-positive follicular dendritic reticulum cells, Ki-M 6-positive macrophages, CD22-positive B-cells, CD1-positive cells, CD3-positive T-cells or HLA-DR-positive cells. Round or elongated MyCs were confined to the thymic medulla and were surrounded by CD3-positive T-cells and CD22-positive B-cells. In MG thymitis MyCs were localized in the vicinity of, but not inside germinal centres (GCs). MyCs were always HLA-DR-negative, but were invariably embedded in a cellular micromilieu with strong HLA-DR expression. A remarkable feature of MG thymitis was that the great majority of MyCs were in intimate contact with intramedullary IDCs. Morphometric studies confirmed that such contacts were significantly less frequent in thymuses from non-myasthenic subjects. This indicates that an IDC-dependent antigen-presenting process for T-cells may actively involve MyCs in MG thymitis.

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.

Immunohistology of adenolymphoma (Warthin's tumour): evidence for a role of vascularization in the organization of the lympho‐epithelial structure

The epithelial, vascular and lymphoid components present in two cases of adenolymphoma (Warthin's tumour) were characterized by immunohistochemistry on frozen sections. Epithelial cells were positive for cytokeratin and keratin and were in close contact with numerous factor VIII related antigen-positive capillary vessels, running parallel to the epithelial basement membrane. The capillary vessels were in communication with high endothelial post-capillary venules which were often lined by HLA-DR-positive endothelial cells. The lymphoid tissue was organized in B-cell and T-cell areas. The B-cell areas consisted of B4+/To15+ B-lymphocytes admixed with DRC-1-positive follicular dendritic reticulum cells. The T-cell areas were mainly populated by T11+/T3+/T4+ lymphocytes admixed with some S-100+/HLA-DR+ interdigitating reticulum cells. Plasma cells and T8+ lymphocytes were more numerous in the proximity of the epithelium. Leu-M3+/PAM-1+ macrophages were scattered singly in the T-cell areas or were arranged in sinus-like structures around the epithelial cysts. The epithelial basement membrane, the vascular membrane and the fibrillar reticular stroma of the lymphoid tissue were immunoreactive for laminin and type IV collagen. The possibility is discussed that the accumulation of lymphoid tissue in Warthin's tumour is modulated by the epithelial cells, perhaps through their ability to organize a peculiar pattern of vascularization.

Enzyme and immunohistochemistry of follicular hyperplasia in AIDS-related lymphadenopathy.

We used a panel of monoclonal and polyclonal antibodies to analyze frozen and paraffin-embedded lymph node biopsy specimens from 25 intravenous drug abusers (IVDA) with acquired immunodeficiency syndrome (AIDS)-related lymphadenopathy histologically characterized by follicular hyperplasia. Our aim was to obtain diagnostic clues to this commonly occurring pattern. Double-labelling immunohistological studies were also performed on selected frozen sections and 13 plastic-embedded specimens were tested by a number of enzyme reactions. Consistent features in IVDA included abnormally high numbers of intrafollicular T-cells, positive for acid phosphatase and beta-glucuronidase, most of which had Leu-2a-positive phenotype; a marked reduction or loss of mantle zone B-cells (positive for surface IgD-IgM and alkaline phosphatase); and disarray of the network of follicular dendritic reticulum cells (DRCs), as revealed with DRC-1 and anti-S-100 protein antibodies or with reaction for 5'-nucleotidase. When present, distinctive intrafollicular clusters of Leu-2a-positive T-cells and mantle zone B-cells were nearly always associated with areas lacking DRCs in some patients. The intrafollicular hypervascularity invariably found in IVDA proved to be of a true capillary nature, as demonstrated by alkaline phosphatase, 5'-nucleotidase, and ATPase reactions. In control tissues, all showing absence of Leu-2a-positive intrafollicular T-cells, most of the above individual changes could be detected, although they were occasional, mild, and never associated within the same follicle. By contrast, combined immunohistological and enzyme histochemical findings in IVDA indicated that in most follicles such changes were marked and very often associated within the same follicle in each case.(ABSTRACT TRUNCATED AT 250 WORDS)

Selective visualization of human dendritic reticulum cells in reactive lymphoid follicles by the immunohistochemical demonstration of the subunit A of factor XIII (F-XIIIa).

Immunohistochemical techniques have been used to localize clotting factor XIII subunit A in human reactive lymphoid follicles. The follicular dendritic reticulum cells (DRCs) were identified by the monoclonal antibodies R4/23 and OKB-7 as well as by their 5'-nucleotidase positivity. Follicular histiocytic reticulum cells (HRCs) were demonstrated by their acid phosphatase and non-specific esterase reactions. Capillaries were selectively visualized by adenosine triphosphatase. The immunohistochemical demonstration of F-XIIIa was preferably carried out in combination with one or two of the above marker techniques, on the same cryostat section. The subunit A of factor XIII is present in follicular DRCs. Their selective immunohistochemical demonstration with antibody against F-XIIIa requires formaldehyde fixation of cryostat sections. Similar fixation, however, is inappropriate for the demonstration of F-XIIIa reactivity of DRCs in paraffin sections. For this purpose, acetic acid-formalin fixation is useful. Follicular HRCs are consistently negative for F-XIIIa, contrary to the F-XIIIa positivity of sinusoidal and interfollicular HRCs. Developmental and functional implications of F-XIIIa reactivity in DRCs and HRCs are suggested.

Immunohistologic analysis of lymphoid cells by a rapid double staining method

The development of monoclonal antibodies has allowed characterization of subpopulations of lymphoid cells and of their in situ distribution in tissues. The feasibility of simulataneous localization of two surface antigens was studied by double staining with monoclonal antibodies to B cells, T cell subsets and follicular dendritic reticulum cells (DRC) using the avidin-biotin-complex (ABC) method in sections of human lymphoid tissues (tonsil, lymph node, spleen) and a non-lymphoid tissue, endometrium. Color mixture was avoided when an additional incubation with avidin-biotin-labeled peroxidase and subsequent development in the respective substrate of the first sequence preceded the second staining sequence using the primary antibodies at optimal concentrations. The antigenic profiles were portrayed by contrasting and distinct colors of the reaction products. It was observed that T lymphocytes of the cytotoxic/suppressor and helper/inducer phenotypes were topographically associated with each other and with B cells in B and T cell areas of lymphoid tissues as well as in lymphocytic aggregates in endometria. Subpopulations of these cells were mantled by processes of DRCs in lymphoid follicles. The findings indicate that the double ABC staining method can be used for simultaneous demonstration of two surface antigens in tissue sections.

Significance of S-100 protein immunostaining in the immunohistological analysis of normal and neoplastic lymphoid tissues--an appraisal.

S-100 protein is a heterogeneous fraction of dimeric polypeptides (alpha and beta subunits) that can exist in different combination forms within the various tissues. Concerning the S-100 protein immunodetection within lymphoid tissue, the heterogeneity of the S-100 antigen, the tissue quality (frozen or paraffin-embedded after treatment with different fixatives) and the treatment of the tissue with different immunostaining methods and antibodies of different nature, all make for inconsistent results obtained in the immunohistological studies reported in the literature. Most of the S-100-positive cells of the lymphoreticular system are dendritic cells involved in the immune response (interdigitating reticulum cells, Langerhans cells, and follicular dendritic reticulum cells), other S-100-positive cells belonging to the mononuclear/phagocytic system. S-100 protein immunostaining may be used as a helpful immunohistological diagnostic clue to certain malignancies of the immune system (follicular center cell lymphomas) on the basis of their specifically related dendritic cell microenvironment. In addition to monoclonal antibodies for the immunophenotypic characterization of dendritic cells and macrophages and to enzyme reactions, the combined use of anti-S-100 antibodies specific for each of the S-100 protein subunits, tested with sensitive procedures, would be a very useful tool in the attempt to classify the proliferative disorders of dendritic cells and macrophages.

Dendritic reticulum cells in reactive lymph nodes and tonsils: an immunohistological study.

There has recently been much interest in the patterns of follicular dendritic reticulum cells (DRC) in pathological lymph nodes, particularly in relation to the phenomenon of DRC break-up (thought to be pathognomonic of AIDS-related lymphadenopathies) and to progressive transformation of germinal centres (as a possible precursor of lymphocyte predominant Hodgkin's disease). In the present study we have immunostained twenty-nine reactive lymph nodes and five tonsils with monoclonal antibody R4/23 (DAKO-DRC) in order to evaluate the frequency of such changes in lymphoid tissue unaffected by AIDS or Hodgkin's disease. Most of the specimens contained typical secondary follicles with clearly defined germinal centres and mantle zones. There were two variants in lymph nodes showing follicular hyperplasia characterized by (i) progressive transformation of germinal centres and (ii) inclusions of nests of small lymphocytes within germinal centres. In each of these types of follicles the compact evenly-distributed meshwork of DRCs, as previously described, was seen. However there were considerable variations in DRC meshwork in each category (the pattern could not be predicted from the morphology) with examples in all three of the DRC break-up previously considered specific for the AIDS related lymphadenopathy. Since none of the lymph nodes and tonsils studied had any known relationship to either Hodgkin's disease or AIDS it is argued that none of the changes in the DRC meshwork observed are specific for these conditions.