Formation of Multinucleated Giant Cells (MGC) in Vitro by Sensitized Macrophages Exposed to Mycobacterium Bovis

Adult rabbits were injected intravenously with heat killed Mycobacterium bovis strain Bacillus Calmette-Guerin (BCG) suspended in oil. As reported previously (1) the rabbits develop a strong infiltration of mononuclear cells which can easily be demonstrated in the lungs 40-50 days post-sensitization. Cells lavaged from these lungs consists of about 82% macrophages, 14% lymphocytes, 3% polymorphonuclear leukocytes and 1% giant cells. Within five to six hours after in vitro cultivation with heat-killed BCG, the macrophages which have already attached to the plastic substrate form clusters apparently by migration. When the mixed population of lavaged cells is cultured for 24 hours many macrophages seem to fuse to form multinucleated giant cells (MGC), whereas cells collected from unsensitized rabbits do not form significant numbers of MGC. A typical MGC containing at least 35 nuclei in the central area of the syncytium is shown in Fig. 1. The mechanism involved in this cell fusion process is not understood.

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Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

Infection and Immunity ◽

10.1128/iai.67.1.395-402.1999 ◽

1999 ◽

Vol 67 (1) ◽

pp. 395-402 ◽

Cited By ~ 37

Author(s):

Annette Gasser ◽

Johannes Möst

Keyword(s):

Mycobacterium Bovis ◽

Giant Cells ◽

Fusion Process ◽

Human Monocytes ◽

Multinucleated Giant Cells ◽

Necrosis Factor Alpha ◽

Human T Cell ◽

Cell Clones ◽

Membrane Bound

ABSTRACT Multinucleated giant cells (MGC), a characteristic feature of tuberculous granulomas, form by fusion of monocytes or macrophages, but little is known about the mechanism of the fusion process itself. Several studies report an indirect effect of mycobacteria, i.e., induction of a soluble lymphocyte-derived fusion factor following stimulation by mycobacteria or mycobacterial products. The aim of our study was to determine whether contact with mycobacteria can induce MGC formation from human monocytes in vitro. Stimulation of monocytes withMycobacterium bovis bacillus Calmette-Guérin (BCG) in combination with cytokine-containing supernatants of herpesvirus saimiri-transformed human T-cell clones (T-SN) led to MGC formation with fusion rates of about 27%. In contrast, stimulation with one component alone induced only low fusion rates of up to 10%. Heat-killed BCG in combination with T-SN induced monocyte fusion to the same extent as live mycobacteria. BCG culture supernatant, BCG lysate, or inert particles in combination with T-SN did not induce MGC formation. Experiments using transwell plates containing a semipermeable membrane revealed that induction of the fusion process is dependent on direct contact of monocytes and mycobacteria. MGC formation induced by BCG plus T-SN could be inhibited by addition of monoclonal antibodies to gamma interferon (but not tumor necrosis factor alpha) as well as to the β chain (CD18) of β2-integrins. These results demonstrate that contact with mycobacteria in combination with cytokine-containing supernatants is able to induce human monocytes to form MGC and that membrane-bound molecules of mycobacteria and monocytes are involved in the fusion process.

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Impact of in vitro evolution on antigenic diversity of Mycobacterium bovis bacillus Calmette-Guerin (BCG)

Vaccine ◽

10.1016/j.vaccine.2014.07.113 ◽

2014 ◽

Vol 32 (45) ◽

pp. 5998-6004 ◽

Cited By ~ 42

Author(s):

Richard Copin ◽

Mireia Coscollá ◽

Efstratios Efstathiadis ◽

Sebastien Gagneux ◽

Joel D. Ernst

Keyword(s):

Mycobacterium Bovis ◽

Bacillus Calmette Guerin ◽

In Vitro Evolution ◽

Antigenic Diversity

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Giant Cell Granuloma of the Temporal Bone: A Case Report With Immunohistochemical, Enzyme Histochemical, and In Vitro Studies

Archives of Pathology & Laboratory Medicine ◽

10.5858/2003-127-1217-gcgott ◽

2003 ◽

Vol 127 (9) ◽

pp. 1217-1220 ◽

Cited By ~ 1

Author(s):

Xue-Fei Tian ◽

Tie-Jun Li ◽

Shi-Feng Yu

Keyword(s):

Giant Cell ◽

Temporal Bone ◽

Mononuclear Cells ◽

Giant Cells ◽

Tartrate Resistant Acid Phosphatase ◽

Giant Cell Granuloma ◽

Ki 67 ◽

The Right ◽

Histochemical Reactivity

Abstract A case of giant cell granuloma (GCG) that occurred in the right temporal bone is reported. The lesion showed histologic features identical to GCG. The multinuclear giant cells (MGCs) in the lesion showed strong reactivity with CD68, but patchy staining for myeloid/histiocyte antigen, α-1-antitrypsin, α-1-antichymotrypsine, and lysozyme. Activity of tartrate-resistant acid phosphatase was also consistently detected in the MGCs. Some of the mononuclear cells of the lesion exhibited similar immunocytochemical and histochemical reactivity as the MGCs. Ki-67 staining, however, was only detected in the mononuclear cells. The MGCs isolated from the lesion presented characteristic morphology of osteoclasts and possessed the ability to excavate bone in vitro. Thus, the MGCs in GCG appeared to express both macrophage- and osteoclast-associated phenotypes. The mononuclear cells were the major proliferative elements in the lesion and a subpopulation of these cells may represent precursors of the MGCs.

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The in vitro immunomodulatory effects of sulfasalazine on human polymorphonuclear leukocytes, mononuclear cells, and cultured glomerular mesangial cells

Life Sciences ◽

10.1016/s0024-3205(00)00708-6 ◽

2000 ◽

Vol 67 (10) ◽

pp. 1149-1161 ◽

Cited By ~ 4

Author(s):

Chang-Youh Tsai ◽

Tsai-Hung Wu ◽

Chia-Li Yu ◽

Chung-Tei Chou

Keyword(s):

Polymorphonuclear Leukocytes ◽

Mononuclear Cells ◽

Mesangial Cells ◽

Glomerular Mesangial Cells ◽

Immunomodulatory Effects ◽

Human Polymorphonuclear Leukocytes

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Mesenchymal stromal cells' secretome enhances osteoclastogenesis but reduces multinucleated giant cells formation in vitro

Bone Reports ◽

10.1016/j.bonr.2020.100371 ◽

2020 ◽

Vol 13 ◽

pp. 100371

Author(s):

Paul Humbert ◽

Julien De Lima ◽

Meadhbh Á. Brennan ◽

Frédéric Blanchard ◽

Pierre Layrolle

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Giant Cells ◽

Multinucleated Giant Cells

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Demonstration of HIV-encoded Proteins in Cultured and in Uncultured CD 4 Positive Mononuclear Cells from Hemophilia Patients Employing Monoclonal Antibodies against p 15, p 24, GP 41, GP 120, and Reverse Transcriptase

10.1055/s-0038-1644683 ◽

1987 ◽

Author(s):

P Wernet ◽

E M Scheider ◽

P Sarin ◽

P Chandra ◽

H H Brackmann ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Reverse Transcriptase ◽

Antigen Processing ◽

Mononuclear Cells ◽

Interleukin 2 ◽

In Vitro Cultivation ◽

Success Rates ◽

Culture Techniques ◽

Gp 120

In the light of the large percentage of hemophilia patients with antibodies to HIV the identification of a specific virus infection in comparison to HIV antibody negative hemophilia patients has reached crucial importance. The low success rates of direct virus culture techniques together with the as yet low AIDS-di-sease rate observed in these patients separate these patients from the other main risk groups. Within this context, we studied the expression of CD3, CD4, CD8, and HLA class II antigens on fixed cells after PHA stimulation and Interleukin 2 propagation as well as on untreated blood mononuclear cells from healthy individuals and from hemophilia patients by fluorescence activated flow cytometry. Monoclonal antibodies thought to be specific for p 15, p 24, GP 41, GP 120, and for reverse transcriptase revealed a certain number of positive cells on all defined subpopulations analysed. From cell specimen of HIV antibody positive hemophilia patients cells with specific HIV antigens could be enriched by in vitro cultivation. Importantly the expression of virus-encoded antigens preceedes a cytopathic effect for several daVs. Current analyses aim at the prognostic relevance of low amounts of such viral HIV proteins selectively detectable by moAbs.directed to either p 24, GP 41, GP 120, and RT. The reliability, high sensitivity and monoclonal antibody dependent specificity of this newly developed method for the demonstration of HIV specific proteins make it applicable also for longitudinal surveys of hemophilia patients to assess a potential state of viremia or virus antigen processing in their mononuclear cells.

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Osseoinduction Evaluation of Hydroxyapatite and Zinc Containing Hydroxyapatite Granules in Rabbits

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.493-494.252 ◽

2011 ◽

Vol 493-494 ◽

pp. 252-257 ◽

Cited By ~ 1

Author(s):

L. Nascimento ◽

M. Medeiros ◽

J. Calasans-Maia ◽

A. Alves ◽

Antonella M. Rossi ◽

...

Keyword(s):

Histological Analysis ◽

Bone Matrix ◽

Fibrous Tissue ◽

Particle Diameter ◽

Inflammatory Cells ◽

Giant Cells ◽

Multinucleated Giant Cells ◽

Ethics Commission

This study investigated the osteoinductive potential of granules of stoichiometric hydroxyapatite (HA) and 0.5% zinc containing hydroxyapatite (ZnHA) in intramuscular (IM) site of rabbit’s abdomen. The biomaterials were both used in granular form, with 75% porosity and particle diameter between 450 and 500μm, sintered at 1100°C. Both materials performed adequately on a multiparametric in vitro cytocompatibility assay, indicating their suitability for in vivo testing. After approval by the Ethics Commission on Teaching and Research in Animals, fifteen rabbits were submitted to general anesthesia, incision and tissue dilatation, and a small site was created for HA (right incision) and ZnHA (left incision) intramuscular implantation. The animals were killed after 2, 4 and 12 weeks for biomaterials and surrounding tissues removal. Histological analysis after 2 weeks revealed the presence of granulation tissue surrounding biomaterials with multinucleated giant cells and no newly formed bone for both materials. After 4 weeks there was fibrous tissue involving the material and few inflammatory cells. Following 12 weeks it was observed the presence of connective tissue surrounding the biomaterial, cellularized enough for the two experimental groups, but it was not observed the presence of bone matrix associated with the biomaterials. We conclude that both biomaterials are cytocompatible and did not present the property of osseoinduction after 12 weeks of implantation.

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