Monocytoid B lymphocytes and epithelioid cell clusters in abscess-forming granulomatous lymphadenitis. With special reference to cat scratch disease.

In order to clarify the appearance of monocytoid B lymphocytes (MBLs) in abscess-forming granulomatous lymphadenitis (AGL) and the relation between AGL and cat-scratch disease (CSD), 48 cases of AGL were studied histologically. MBLs were present in about 50% of AGL cases. Warthin-Starry (WS) silver stain-positive bacteria, which are the causative agent of CSD, were present in 52.4% of AGL cases with MBLs and 59.2% of AGL cases without MBLs. The appearance of MBLs in AGL was not related to various clinical features, including disease interval from initial lymphadenopathy to lymph node biopsy. Histologically, epithelioid cell clusters appeared in about 70% of MBL-positive AGL cases, but were not observed in MBL-negative AGL. Therefore, a close interaction between MBLs and epithelioid cells in AGL is suggested, and we emphasize that the histological features of some AGL cases resemble those of toxoplasmic lymphadenitis.     

Monocytoid B Lymphocytes and Epithelioid Cell Clusters in Abscess-forming Granulomatous Lymphadenitis With Special Reference to Cat Scratch Disease

In order to clarify the appearance of monocytoid B lymphocytes (MBLs) in abscess-forming granulomatous lymphadenitis (AGL) and the relation between AGL and cat-scratch disease (CSD), 48 cases of AGL were studied histologically. MBLs were present in about 50% of AGL cases. Warthin Starry (WS) silver stain-positive bacteria, which are the causative agent of CSD, were present in 52.4% of AGL cases with MBLs and 59.2% of AGL cases without MBLs. The appearance of MBLs in AGL was not related to various clinical features, including disease interval from initial lymphadenopathy to lymph node biopsy. Histologically, epithelioid cell clusters appeared in about 70% of MBL positive AGL cases, but were not observed in MBL negative AGL. Therefore, a close interaction between MBLs and epithelioid cells in AGL is suggested, and we emphasize that the histological features of some AGL cases resemble those of toxoplasmic lymphadenitis.     

Cytologic diagnosis of cat scratch disease (CSD) by fine-needle aspiration

Cat scratch disease (CSD) is usually a benign, self-limited lymphadenitis, characterized by suppurative granulomas. It can, however, produce a wide spectrum of clinical symptoms and cytologic changes and be the source of diagnostic dilemmas. Identification of pleomorphic bacilli (PB) with silver impregnation stains aids in the diagnosis, but this has not been well documented in cytologic preparations or in cases without the classic morphologic changes. We reviewed 13 aspirations from eight patients (aged 13–36 yr) occurring over a 15 mo time period, all clinically or cytologically suspicious for CSD. Sites included: axilla (6), parotid (3), epitrochlear (1), neck (1), submental (1), and intraclavicular (1) nodes. Neoplasia was initially suspected clinically in 38% of the cases. All but two patients had cat exposure on subsequent interview. The cytologic differential included bacterial abscess and lymphoproliferative disorders in 31%. Neither granulomas nor suppurative inflammation were seen in all cases. Changes included: granulomas (77%), PMNs (62%), dispersed epithelioid histiocytes (46%), and suppurative granulomas (38%). A modified silver stain (Modified Steiner, Sigma Diagnostics, St. Louis, MO) was performed on all specimens. Silver positive organisms were seen in 69% of cases and were not limited to those preparations with suppurative granulomas. Fine-needle aspiration biopsy (FNAB) is an effective method for diagnosing CSD despite its heterogeneous appearance; and, when combined with clinical information and silver staining, may obviate the need for excision. © 1995 Wiley-Liss, Inc.     

Plasmacytoid monocytes in cat scratch disease with special reference to the histological diversity of suppurative lesions

It has been suggested that plasmacytoid monocytes (PMOs) play an essential role in T-cell-dependent immune response. Indeed, numerous PMOs are found in close topographical association with epithelioid cell granulomas in hypersensitivity-type granulomas, such as tuberculosis and sarcoidosis. The key pathologic process in cat scratch disease (CSD) usually involves a B-cell-associated granulomatous reaction. Histologically, CSD appears to exhibit a histopathologic diversity, including suppurative lesions without epithelioid cell granulomas (early lesion), in which the microabscesses were surrounded by monocytoid B-cells (MBCs), suppurative granulomas containing MBCs (intermediate lesion), and suppurative granulomas without MBCs (late lesion). However, the presence or absence of PMO in CSD has not been studied previously. We examined 14 cases of CSD. In early lesions, numerous clusters of PMO were detected in the MBCs. In intermediate lesions, both MBCs and PMOs were found to be decreased in number, while late lesions contained no or only a few MBCs and PMOs. Overall, these findings suggest that PMOs may play a role in MBC-associated granulomatous response and in hypersensitivity granulomatous response. Moreover, the association with MBCs and PMOs indicates a functional relationship of MBCs with PMOs in the formation of suppurative lesions in CSD.     

Atypical large plasma cells in lymph node granulomas in cat-scratch disease

Summary A histological variant of plasma cells found in the granulomas of cat-scratch disease (CSD) lymphadenitis is reported. Though the lesion shows the typical features of suppurative granulomatous lymphadenitis, many atypical giant cells which have abundant basophilic cytoplasm and bizarre nuclei with occasional multi-nucleated forms are noted among epithelioid histiocytes. The diagnosis of CSD lymphadenitis was confirmed by comparing clinical, histopathological, and histochemical (Warthin-Starry silver impregnation stain) studies on lymph node sections from five cases with features typical of the disease. Histochemical (methyl green-pyronine stain) and immunohistochemical examination provided several lines of evidence indicating that the atypical giant cells in our case were plasmacytic and confirmed that its proliferation was reactive, not neoplastic. Multinucleated giant cells were also occasionally present in the other five cases, but they had histological and immunohistochemical features of Langhans' type giant cells. We stress the importance of distinguishing such atypical large plasma cells from neoplastic cells.     

The role of macrophage colony-stimulating factor in hepatic glucan-induced granuloma formation in the osteopetrosis mutant mouse defective in the production of macrophage colony-stimulating factor.

To elucidate the effects of macrophage colony-stimulating factor (M-CSF) on Kupffer cells and monocyte/macrophages in hepatic granuloma formation, we examined granulomas produced by glucan injection in the liver of osteopetrotic mice and littermates with or without M-CSF administration. In the osteopetrotic mice, monocytes were deficient in peripheral blood, and their number did not increase after glucan injection. Hepatic granulomas were formed in the osteopetrotic mice by glucan injection without a supply of blood monocytes. During this process, M-CSF-independent Kupffer cells proliferated, particularly before the granuloma formation, clustered in the hepatic sinusoid, and transformed into epithelioid cells and multinuclear giant cells. In the M-CSF-treated osteopetrotic mice, glucan injection induced an increase in the number of blood monocytes and formed hepatic granulomas at a nearly similar degree to that of littermate mice. Thus, it is concluded that neither monocytes nor M-CSF are necessary for granuloma formation. In contrast, Kupffer cells play a crucial role as granulomas develop in M-CSF-uninjected osteopetrotic mice.     

The role of Kupffer cells in glucan-induced granuloma formation in the liver of mice depleted of blood monocytes by administration of strontium-89

In order to elucidate the role of Kupffer cells in granuloma formation in the liver of mice under a condition of severe monocytopenia induced by administration of strontium-89, granulomas were produced by particulate glucan injection and examined histopathologically, immunohistochemically, by [3H]thymidine autoradiography, and in culture experiments. Hepatic granulomas were smaller, less numerous, and more irregularly shaped in the monocytopenic mice than in the control mice. The granulomas were composed of multinuclear giant cells, epithelioid cells, Kupffer cells, and T lymphocytes, but not monocytes or granulocytes. Kupffer cells were heavily labeled with [3H]thymidine in the monocytopenic mice, particularly just before the stage of granuloma formation, and then clustered in the liver sinusoids. At 8 days, they formed granulomas, transformed into epithelioid cells, and transformed further into multinuclear giant cells. Although the culture of liver cell suspensions prepared from the livers of monocytopenic mice sustained diffuse proliferation of macrophages on a monolayer of mouse stromal cell line (ST2), no monocyte/macrophage colonies were formed. From these results, it is reasonable to conclude that Kupffer cells alone are activated in a condition without a supply of monocytes from peripheral blood; proliferate and cluster in the hepatic sinusoids; transform into peroxidase-negative macrophages, epithelioid cells, and multinuclear giant cells; and participate in granuloma formation in loco together with T lymphocytes.     

Macrophage Polarization Drives Granuloma Outcome during Mycobacterium tuberculosis Infection

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), induces formation of granulomas, structures in which immune cells and bacteria colocalize. Macrophages are among the most abundant cell types in granulomas and have been shown to serve as both critical bactericidal cells and targets for M. tuberculosis infection and proliferation throughout the course of infection. Very little is known about how these processes are regulated, what controls macrophage microenvironment-specific polarization and plasticity, or why some granulomas control bacteria and others permit bacterial dissemination. We take a computational-biology approach to investigate mechanisms that drive macrophage polarization, function, and bacterial control in granulomas. We define a “macrophage polarization ratio” as a metric to understand how cytokine signaling translates into polarization of single macrophages in a granuloma, which in turn modulates cellular functions, including antimicrobial activity and cytokine production. Ultimately, we extend this macrophage ratio to the tissue scale and define a “granuloma polarization ratio” describing mean polarization measures for entire granulomas. Here we coupled experimental data from nonhuman primate TB granulomas to our computational model, and we predict two novel and testable hypotheses regarding macrophage profiles in TB outcomes. First, the temporal dynamics of granuloma polarization ratios are predictive of granuloma outcome. Second, stable necrotic granulomas with low CFU counts and limited inflammation are characterized by short NF-κB signal activation intervals. These results suggest that the dynamics of NF-κB signaling is a viable therapeutic target to promote M₁ polarization early during infection and to improve outcome.     

Cat-scratch disease in a patient with AIDS

A case of cat-scratch disease (CSD) in a patient with acquired immunodeficiency syndrome (AIDS) is reported. The lymph node pathologic characteristics were altered from those usually seen with CSD, showing clusters of vacuolated macrophages admixed with pycnotic nuclear debris instead of the usual suppurative granulomas. Evidence for the diagnosis was provided by Warthin-Starry stain and electron microscopic demonstration of the presumed CSD bacillus. Empiric treatment with antibiotics brought about clinical improvement. This case demonstrates the altered lymph node pathologic characteristics with CSD that may occur in a patient with AIDS.     

Live Mycobacterium avium subsp. paratuberculosis and a killed-bacterium vaccine induce distinct subcutaneous granulomas, with unique cellular and cytokine profiles

Type II (lepromatous) granulomas are characterized by a lack of organization, with large numbers of macrophages heavily burdened with bacilli and disorganized lymphocyte infiltrations. Type II granulomas are a characteristic feature of the enteric lesions that develop during clinical Mycobacterium avium subsp. paratuberculosis infection in the bovine. Considering the poor organization and function of these granulomas, it is our hypothesis that dendritic cell (DC) function within the granuloma is impaired during initial infection. In order to test our hypothesis, we used a subcutaneous M. avium subsp. paratuberculosis infection model to examine early DC function within M. avium subsp. paratuberculosis-induced granulomas. In this model, we first characterized the morphology, cellular composition, and cytokine profiles of subcutaneous granulomas that develop 7 days after subcutaneous inoculation with either vaccine or live M. avium subsp. paratuberculosis. Second, we isolated CD11c(+) cells from within granulomas and measured their maturation status and ability to induce T-cell responses. Our results demonstrate that M. avium subsp. paratuberculosis or vaccine administration resulted in the formation of distinct granulomas with unique cellular and cytokine profiles. These distinct profiles corresponded to significant differences in the phenotypes and functional responses of DCs from within the granulomas. Specifically, the DCs from the M. avium subsp. paratuberculosis-induced granulomas had lower levels of expression of costimulatory and chemokine receptors, suggesting limited maturation. This DC phenotype was associated with weaker induction of T-cell proliferation. Taken together, these findings suggest that M. avium subsp. paratuberculosis infection in vivo influences DC function, which may shape the developing granuloma and initial local protection.     

Mycobacterial granulomas: keys to a long-lasting host-pathogen relationship

Chronic infection with mycobacteria is controlled by the formation of granulomas. The failure of granuloma maintenance results in reactivation of disease. Macrophages are the dominant cell type in granulomas, but CD4+ T cells are the master organizers of granuloma structure and function. Recent work points to an unrecognized role for nonspecific T cells in maintaining granuloma function in the chronic phase of infection. In addition, it has become clear that mycobacteria and host T cells collaborate in formation of granulomas. Further understanding of how nonspecific T cells contribute to granuloma formation, as well as how bacteria and T cells maintain a harmonious relationship over the life of the host, will facilitate the development of new strategies to treat mycobacterial disease.     

Comparison of mycobacterial granulomas guinea-pig lymph nodes

A study was made of mycobacterial-induced granulomas in guinea-pig lymph nodes. Live BCG (Pasteur) induced a granuloma containing epithelioid cells while Cobalt irradiated Mycobacterium leprae induced a granuloma comprised of phagocytic macrophages. The granulomas were quantitated by measurement of lymph node weight and the areas of infiltration in histological sections. The time course of granuloma formation induced by Co-irradiated M. leprae was very different from the time course of the granuloma formation induced by BCG. Collagen synthesis assessed by incorporation of ¹⁴C-proline into collagenase sensitive protein was greater in lymph nodes draining the site of injection of Co-irradiated BCG than those draining the site of injection of Co-irradiated M. leprae during the first 10 weeks. Collagen synthesis was delayed in the nodes from animals injected with live BCG for at least 10 weeks. Single cell suspensions of draining lymph nodes containing granulomas consisted of lymphocytes and large cells (epithelioid cells and macrophages). A high proportion of the large cells were found to be non-adherent in the live BCG-induced epithelioid cell granuloma. In contrast, M. leprae-induced granulomas contained a high percentage of adherent large cells. In both the granulomas, the majority of large cells were esterase positive and showed the presence of fibronectin. Most of the large cells in the granulomas did not carry receptors for the Fc component of IgG or the C3 component of complement and did not exhibit peroxidase activity.     

Th2-type granuloma development in acute murine schistosomiasis is only partly dependent on CD4+ T cells as the source of IL-4

Schistosome granulomas produce IL-4, important for Th2 granuloma expression. We defined the origins of IL-4 within these granulomas and the role of IL-4-producing CD4(+) T cells in Th2 granuloma development. Dispersed granuloma cells spontaneously produced IL-4 independently of T cells, whereas IL-5 production was T cell dependent. Granuloma IL-4 mRNA localized to the non-T cells and IL-5 to T cells. Granuloma CD4(+) T and NK cells, but not B cells produced IL-4 and IL-5 in vitro. B cell-/- mice generated Th2 granulomas that produced IL-4 and IL-5 normally. Granuloma eosinophils expressed no IL-4 or IL-5 mRNA. Granulomas in WWv mast cell-deficient mice lacked mast cells. The dispersed granuloma cells from WWv mice released IL-4 only after T cell stimulation, suggesting that mast cells influenced the constitutive component of IL-4 production. Rag-1 animals (T/B/NK T cell deficient) given schistosomiasis after reconstitution with splenocytes from naive mice produced Th2 granulomas. Mice reconstituted to create selective CD4(+) T cell IL-4 knockout animals developed eosinophilic granulomas that made IL-4. Thus, granulomas contain several cell types that produce IL-4. Mast cells are not needed to form Th2 granulomas, but influence IL-4 release. Th2 granuloma development in schistosomiasis is only partly dependent on IL-4-producing CD4(+) T cells.     

Virulence of Paracoccidioides brasiliensis isolates can be correlated to groups defined by random amplified polymorphic DNA analysis.

Fifteen Paracoccidioides brasiliensis isolates were discriminated by the RAPD analysis into two groups with only 17% of genomic identity. The ability of P. brasiliensis isolates to invade tissues was studied in an experimental model using susceptible B10.A mice. The analysis was performed according to the severity of the lesions including the number and size of the granuloma, the number and dissemination of fungi to different organs. The isolates from two RAPD groups demonstrated a marked difference in their virulence patterns for B10.A mice. The isolates Pb S, 662, Bt and 166 (group I) elicited localized infection restricted to the liver showing compact epithelioid granuloma with few fungi in the early post-infection period (slightly virulent). On the other hand the isolates Pb 01 and 7455 (group II) elicited a disseminated infection with a mixed suppurative and looser granulomatous inflammation, showing extensive areas of necrosis and large numbers of viable fungal cells (highly virulent). These results are strong evidence for correlation between RAPD patterns and the virulence degree of P. brasiliensis.     

伴反应性肉芽肿的乳腺癌

目的：研究伴反应性肉芽肿的乳腺癌的临床病理特点。方法：观察5例伴反应性肉芽肿的乳腺癌的形态学改变。结果：5例病人都没有全身性肉芽肿性疾病。癌组织内均可见非干酪性肉芽肿,肉芽肿沿癌细胞分布,其内有朗汉斯多核巨细胞区域性淋巴结内没有反应性肉芽肿。结论：伴反应性肉芽肿的乳腺癌需要和乳腺伴破骨细胞样巨细胞的瘤、伴破骨细胞样巨细胞的化生性癌、肉芽肿性小叶性乳腺炎、间质反应性巨细胞、伴结节病或伴结核病的癌鉴别。     

Direct evidence for granuloma-inducing activity of interleukin-1. Induction of experimental pulmonary granuloma formation in mice by interleukin-1-coupled beads.

Pulmonary granulomas were induced in BALB/c mice by the intratracheal injection of Sephadex G-50 and latex beads. Very large granulomas developed around Sephadex G-50 beads. Minimal inflammation was produced in mice given latex beads. Aqueous extracts prepared from pulmonary granuloma lesions induced in mice by Sephadex G-50 beads contained high levels of interleukin-1 (IL-1) activity but not interleukin-2 (IL-2) activity. IL-1 activity in the extracts correlated with granuloma size. In a subsequent step, large granulomas were induced by the intratracheal injection of Sepharose 4B beads coupled to fractions of the extracts containing IL-1 activity (ie, granuloma-derived IL-1) prepared from Sephadex G-50-induced granulomatous lungs. In addition, large granulomas were induced by the intratracheal injection of recombinant IL-1-coated Sepharose 4B beads. In contrast, very small granulomas were seen when IL-2-coated or plain Sepharose 4B beads were injected into mice. These results indicate that IL-1 participates in the induction and/or expression of granulomas.     

High sensitivity of transgenic mice expressing soluble TNFR1 fusion protein to mycobacterial infections: Synergistic action of TNF and IFN-γ in the differentiation of protective granulomas

To investigate the role of tumor necrosis factor (TNF) in protective immune responses to Mycobacterium tuberculosis and M. bovis Bacillus Calmette Guérin (BCG), we have used transgenic mice unable to use TNF because of the expression of high amounts of a soluble TNF receptor (R) type I (sTNFR1) fusion protein, and studied resistance of these mice to infection by lethality assays, evaluation of bacterial recovery and histologic examination. These mice showed a strongly increased sensitivity to M. tuberculosis and BCG infections, with bacterial overgrowth and marked inhibition of macrophage differentiation within granulomas; after M. tuberculosis infection, this resulted in extensive lesions of caseous necrosis in the lung. To explore the respective roles of TNF and interferon (IFN)-γ in resistance to BCG and granuloma differentiation, controls and sTNFR1-transgenic mice were compared to IFN-γR mutant mice and mice double defective in TNF and IFN-γ activity (obtained by crossing transgenic and mutant mice). The three groups of deficient mice showed a strongly enhanced susceptibility to BCG infection, with the following decreasing order of sensitivity between groups: TNF + IFN-γ → TNF → IFN-γ-deficient mice. The hepatic granulomas of IFN-γR mutant mice were small and contained eosinophils but few differentiated macrophages; compared to those of sTNFR1-transgenic mice, acid-fast bacilli were less numerous within the macrophages. Granulomas of double-deficient mice were strikingly different by their very large size and cellular content, made up large numbers of polymorphonuclears, eosinophils, and cells undergoing apoptosis, but without detectable differentiated macrophages; acid-fast bacilli were spread in the lesions. These studies show the essential role of both TNF and IFN-γ in the development, during mycobacterial infections, of protective granulomas containing highly differentiated macrophages capable of destroying ingested bacteria, and emphasize that these two cytokines act synergistically in granuloma formation.     

Established T(H1) granulomatous responses induced by active Mycobacterium avium infection switch to T(H2) following challenge with Schistosoma mansoni

Mycobacterium avium established a systemic infection with granulomatous inflammation in mice. Mice chronically infected with M. avium and subsequently co-infected with Schistosoma mansoni developed additional, but morphologically distinct, hepatic granulomas. Schistosome eggs were not deposited in the spleen, and splenic granulomas in co-infected mice contained mycobacteria. In complete contrast to the T(H1) cytokine pattern observed with granuloma lymphocytes from M. avium-infected mice, granuloma lymphocytes from co-infected mice stimulated with PPD elaborated IL-4, but not IFN-gamma. Furthermore, mycobacterial granulomas in concurrently infected mice contained large numbers of eosinophils, a feature never seen in granulomas of M. avium-infected mice. Serum IgG1 and IgE levels in concurrently infected mice were significantly higher, but IgG2a levels significantly lower, than those in M. avium-infected mice, further evidence that the T(H1) component induced by M. avium is modulated subsequent to co-infection with S. mansoni. The dominance of the T(H2) response observed in this model could have clinical implications in areas where parasites and mycobacteria co-exist.     

Immunohistochemical and ultrastructural studies on histiocytosis in children

Thirty-four cases of eosinophilic granulomas, 18 cases of diffuse histiocytosis-X, 2 cases of Letterer-Siwe-like syndrome with immunodeficiency, 4 cases of malignant histiocytosis and virus associated hemophagocytic syndrome were studied. On paraffin section, S100 protein, lysozyme, alpha-1-anti-trypsin, alpha-1-antichymotrypsin, alpha-2-macroglobulin, Transferrin, Ferritin, peanuts agglutinin, Concanavalin-A, and dolichos biflorus associated antigen were stained by the immunoperoxidase method. In a few fresh materials, T-cell subpopulation by use of monoclonal antibodies (OKT-3, 4, 6, and OK-M1) was examined by the immunoperoxidase method. Two types of Langerhans' cells were found, one is positive for Ferritin and alpha-2-macroglobulin in diffuse histiocytosis-X cells, and another is negative for them in both eosinophilic granulomas. Diffuse histiocytosis-X cell resembled the transformed type of Langerhans cell more than eosinophilic granuloma cells in cellular differentiation. It seemed that the term prolangerhans' cell proliferation disorder might be responsible for it.     

In situ analysis of B7-2 costimulatory,major histocompatibility complex class II,and adhesion molecule expression in schistosomal egg granulomas.

Immunopathological damage in schistosomiasis consist of egg granuloma formation, which is a hypersensitivity reaction mediated by antigen-specific CD4+ T helper (Th) lymphocytes. Th cells are dependent on accessory cell-bound co-stimulatory signals for activation. The B7 molecules provide critical costimulation, as in their absence T cells are rendered unresponsive. We investigated the kinetics of B7-2 molecule expression in schistosomal egg granulomas by immunocytochemical analysis in situ. We also monitored major histocompatibility complex (MHC) class II, intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) expression, as well as the presence of macrophages, T cells, and B cells. B7-2 expression was elevated in cells of the large granulomas of the acute (6-1/2 week) infection, and sharply declined thereafter. MHC class II expression was similarly elevated in the acute granulomas, but in contrast to B7-2, remained relatively constant throughout 12 1/2 weeks of infection. Moreover, ICAM-1 and VCAM-1 were expressed in acute and chronic granulomas. However, whereas ICAM-1 was constitutively expressed in normal liver, VCAM-1 was dramatically induced in the livers upon onset of disease. The findings suggest that the acute granulomas are rich in accessory cells with overall phenotypic configurations that support Th-cell activation, although at subsequent times, granulomas contain increasing numbers of B7-poor accessory cells capable of inducing Th-cell unresponsiveness. Thus, cellular interactions in early granulomas may be able to amplify egg-induced immunopathology, whereas interactions taking place in succeeding granulomas appear to precipitate its down-regulation.