35 results on '"Ferrarini, M"'
Search Results
2. CD5+ B cells with the features of subepithelial B cells found in human tonsils.
- Author
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Dono M, Burgio VL, Colombo M, Sciacchitano S, Reverberi D, Tarantino V, Cutrona G, Chiorazzi N, and Ferrarini M
- Subjects
- Adolescent, B-Lymphocyte Subsets metabolism, B-Lymphocytes cytology, CD5 Antigens metabolism, Cells, Cultured, Child, Child, Preschool, Flow Cytometry, Fluorescent Antibody Technique, Genes, Immunoglobulin, Humans, Immunoglobulin Heavy Chains genetics, Immunoglobulin Variable Region genetics, Immunohistochemistry, Lymphocyte Subsets immunology, Mutation, Palatine Tonsil metabolism, Polymerase Chain Reaction, B-Lymphocyte Subsets immunology, B-Lymphocytes immunology, CD5 Antigens immunology, Palatine Tonsil immunology
- Abstract
This study describes a CD5+ B cell that differs from the majority of the CD5+ B cells from human tonsils. This cell, isolated from in vivo activated B cells, expressed activation markers and featured a CD23-, IgMhigh, IgDlow surface phenotype, responded to T cell-independent type-2 antigens in vitro, and was detected in the subepithelial (SE) areas, the tonsil equivalent of the splenic marginal zone (MZ). Most of the cells utilized unmutated Ig VH genes, although cells with mutated genes also were found, a finding confirmed by single-cell studies. Mutated sequences were more frequent in suspensions enriched for CD27+ cells. Repeated VDJ gene sequences were observed in different molecular clones from the same cell suspension, suggesting in situ expansion. These CD5+ B cells seem to share features with previously characterized tonsil CD5- SE B cells and differ from the majority of tonsil CD5+ B cells, which have the surface phenotype of follicular mantle B cells, lack activation markers, do not respond to T cell-independent antigens, and utilize unmutated VH genes. These data are discussed considering the present views on the origin of B cell subset populations and the relationships between MZ and B1 cells.
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- 2007
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3. B lymphocytes in humans express ZAP-70 when activated in vivo.
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Cutrona G, Colombo M, Matis S, Reverberi D, Dono M, Tarantino V, Chiorazzi N, and Ferrarini M
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- ADP-ribosyl Cyclase 1 immunology, B-Lymphocytes cytology, B-Lymphocytes enzymology, CD5 Antigens immunology, Cells, Cultured, Germinal Center cytology, Germinal Center immunology, Humans, Immunoglobulin D immunology, Killer Cells, Natural cytology, Killer Cells, Natural enzymology, Killer Cells, Natural immunology, Leukemia, Lymphocytic, Chronic, B-Cell enzymology, Leukemia, Lymphocytic, Chronic, B-Cell immunology, Leukemia, Lymphocytic, Chronic, B-Cell pathology, Membrane Glycoproteins immunology, Palatine Tonsil cytology, Palatine Tonsil immunology, T-Lymphocytes cytology, T-Lymphocytes enzymology, T-Lymphocytes immunology, ZAP-70 Protein-Tyrosine Kinase biosynthesis, B-Lymphocytes immunology, Gene Expression Regulation, Enzymologic immunology, Lymphocyte Activation immunology, ZAP-70 Protein-Tyrosine Kinase immunology
- Abstract
ZAP-70 is a protein tyrosine kinase initially found in T and NK cells. Recently, this important signaling element was detected in leukemic B cells from a subgroup of patients with B cell chronic lymphocytic leukemia (B-CLL). In this study, ZAP-70 was detected in normal B cells from human tonsils, but not from peripheral blood. The cDNA sequence of B cell ZAP-70 was the same as that in T cells. Germinal center B cells and plasma cells had a substantial proportion of ZAP-70+ cells, while memory and follicular mantle B cells, which contain low numbers of activated B cells, expressed relatively little ZAP-70. A cell fraction of IgD+, CD38+ B cells, which are comprised of many in vivo activated B cells, exhibited the highest levels of ZAP-70. Density gradient fractionation of tonsil B cells confirmed that ZAP-70 was not expressed by resting B cells, but was expressed by buoyant, in vivo activated B cells. In these B cells, the expression of ZAP-70 correlated with that of CD38 and not with that of CD5, a hallmark of B-CLL cells. B-CLL cells are activated cells and their ZAP-70 expression reflects a normal property of activated B cells populations rather than a neoplastic aberration.
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- 2006
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4. Maintenance of B lymphocyte-related clones in the cerebrospinal fluid of multiple sclerosis patients.
- Author
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Colombo M, Dono M, Gazzola P, Chiorazzi N, Mancardi G, and Ferrarini M
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- Amino Acid Sequence, Base Sequence, Gene Rearrangement, Humans, Immunoglobulin Heavy Chains chemistry, Immunoglobulin Variable Region chemistry, Longitudinal Studies, Molecular Sequence Data, Time Factors, B-Lymphocytes immunology, Immunoglobulin Heavy Chains genetics, Immunoglobulin Variable Region genetics, Multiple Sclerosis cerebrospinal fluid, Multiple Sclerosis immunology
- Abstract
A longitudinal study of Ig V gene segments utilized by B cells from the cerebrospinal fluid (CSF) of two patients with multiple sclerosis (MS) was carried out using RT-PCR methodologies. One patient with a relapsing-remitting (RR)-MS was investigated at onset and at relapse, 1 year later. A patient with secondary-progressive (SP)-MS was tested 9 and 13 years after disease onset. Sequence analyses of V(H)DJ(H) segments bearing V(H)3 and V(H)4 that were obtained from Cgamma cDNA genes demonstrated a substantial proportion of shared clones in the samples taken at different times; these clones were identical or closely related, i.e. had the same third complementary determining region (CDR) of the H chain variable region gene (HCDR3) with different mutations in the V(H) segment. Collectively, these data demonstrate that in MS patients there is a strong selective pressure, which could be exerted by antigen (or autoantigen) stimulation, for the maintenance and partial diversification of certain V(H)DJ(H) Cgamma sequences.
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- 2003
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5. CD30 ligation differentially affects CXCR4-dependent HIV-1 replication and soluble CD30 secretion in non-Hodgkin cell lines and in gamma delta T lymphocytes.
- Author
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Biswas P, Mantelli B, Delfanti F, Ferrarini M, Poli G, and Lazzarin A
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- Humans, Lymphoma, Non-Hodgkin metabolism, NF-kappa B metabolism, Up-Regulation, HIV-1 metabolism, Ki-1 Antigen metabolism, Receptors, Antigen, T-Cell, gamma-delta metabolism, Receptors, CXCR4 metabolism, T-Lymphocytes metabolism
- Abstract
We studied whether signaling through CD30, a member of the TNF receptor family, affected acute infection with HIV-1, encompassing its entire replicative cycle. Several non-Hodgkin cell lines, targets of CXCR4-dependent (X4) HIV-1 infection, were positive for CD30 expression. CD30 ligation induced up-regulation of viral replication only in certain CD30+ cell lines. Enhancement of X4 virus replication by CD30 engagement inversely correlated with both CD30 surface density and constitutive NF-kappaB activation. Conversely, expression of CD30, but not of other members of the TNF receptor family, was proportional to constitutive NF-kappaB binding. Concomitantly, secretion of soluble (s) CD30 increased in all cell lines by CD30 ligation. sCD30 release was enhanced by engagement of CD30 alone and, to a greater extent, by co-engagement of CD3 also in primary gamma delta T lymphocytes, along with complementary modulations of their surface CD30 expression. sCD30-containing supernatant specifically inhibited HIV-1 expression induced by CD30 engagement in chronically infected ACH-2 T cells; thus sCD30 may act as a negative feed-back molecule. In conclusion, we have delineated novel features of CD30 biology and underline the peculiar link of CD30 expression to constitutive NF-kappaB activation which is pivotal to both HIV replication and cell survival.
- Published
- 2003
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6. Double-edged effect of Vgamma9/Vdelta2 T lymphocytes on viral expression in an in vitro model of HIV-1/mycobacteria co-infection.
- Author
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Biswas P, Ferrarini M, Mantelli B, Fortis C, Poli G, Lazzarin A, and Manfredi AA
- Subjects
- Apoptosis, Cell Division, Cell Line, Flow Cytometry, HIV Infections virology, HIV-1 physiology, Humans, Interferon-gamma metabolism, Models, Biological, Mycobacterium Infections immunology, Organophosphorus Compounds pharmacology, Receptors, Antigen, T-Cell metabolism, T-Lymphocytes drug effects, T-Lymphocytes metabolism, Tumor Necrosis Factor-alpha metabolism, U937 Cells, Virus Replication, fas Receptor metabolism, Gene Expression Regulation, Viral, HIV Infections complications, HIV Infections immunology, HIV-1 genetics, Hemiterpenes, Mycobacterium Infections complications, Receptors, Antigen, T-Cell immunology, T-Lymphocytes immunology
- Abstract
A reciprocal influence exists between mycobacteria and HIV: HIV-infected individuals are more susceptible to mycobacterial infections and, on the other hand, mycobacterial infection results inacceleration of HIV disease progression. Vgamma9/Vdelta2 T lymphocytes are known to participate in the defense against intracellular pathogens, including Mycobacterium tuberculosis. Indeed, they kill mycobacteria-infected macrophages and, upon recognition of mycobacterial Ag, release TNF-alpha and IFN-gamma, which are also up-regulators of HIV expression. To assess whether mycobacteria-activated gamma delta T lymphocytes contribute to the enhancement of HIV replication, we established an in vitro model mimicking HIV and mycobacteria co-infection with the latently HIV-infected promonocytic U1 cell line and Vgamma9/Vdelta2 peripheral lymphocytes stimulated with mycobacterial Ag. gamma delta T cell activation determined two distinct, but connected effects, namely U1cell death and HIV expression. Both effects were mainly mediated by release of TNF-alpha and IFN-gamma from activated gamma delta lymphocytes, although Fas-FasL interaction also contributed to U1 apoptosis. The final outcome on U1 survival, and thus, on HIV expression, highly depended on mycobacterial Ag concentration coupled to the differential secretory potency of gamma delta cells. In particular, the induction of viral expression prevailed at low Ag concentration and with lower cytokine production by mycobacteria-activated gamma delta cells. Notably, during the course of HIV infection, Vgamma9/Vdelta2 lymphocytes are reported to be functionally impaired and may thus indirectly influence the progression of HIV disease. In addition, a predominant inhibition of viral replication was encountered when mycobacteria-activated gamma delta T cells were co-cultured with primary HIV-infected macrophages. Thus, we suggest that specific recognition of mycobacterial Ag by gamma delta T lymphocytes in co-infected individuals may modulate viral replication through the complex array of soluble factors released.
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- 2003
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7. Skewing of cytotoxic activity and chemokine production, but not of chemokine receptor expression, in human type-1/-2 gamma delta T lymphocytes.
- Author
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Dagna L, Iellem A, Biswas P, Resta D, Tantardini F, Fortis C, Sabbadini MG, D'Ambrosio D, Manfredi AA, and Ferrarini M
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- Cell Differentiation, Humans, Interferon-gamma biosynthesis, Interleukin-4 biosynthesis, Ki-1 Antigen analysis, Membrane Glycoproteins biosynthesis, Organophosphorus Compounds immunology, Perforin, Pore Forming Cytotoxic Proteins, U937 Cells, Chemokines biosynthesis, Cytotoxicity, Immunologic, Hemiterpenes, Receptors, Antigen, T-Cell, gamma-delta analysis, Receptors, Chemokine analysis, Th1 Cells immunology, Th2 Cells immunology
- Abstract
Human Vgamma9/Vdelta2(+) T lymphocytes participate in the immune response against intracellular pathogens through the secretion of type-1 cytokines and chemokines and by killing of infected cells. Little is known of the effects by type-2 differentiation of gamma delta cells on these functions. Here, we report that bona fide naive cord blood-derived gamma delta lymphocytes expanded in vitro with the mycobacterial antigen isopentenyl pyrophosphate (IPP) can be differentiated as either type-1 or type-2 cells, in the presence of an appropriate cytokine milieu. Instead, peripheral gamma delta cells from PPD-negative healthy adults displayed a type-1 cytokine profile, i.e. IPP-stimulated secretion of IFN-gamma, but not of IL-4 and IL-10. Moreover, they released the macrophage inflammatory protein (MIP)-1beta, but not IL-8 nor the Th2 chemoattractants I-309 and TARC (thymus and activation-regulated chemokine). This cytokine profile was not significantly affected by in vitro culture in Th2 polarizing conditions. Only in one case out of seven were peripheral gamma delta cells fully differentiated to type-2 lymphocytes, characterized by sustained IL-4 and IL-10 production, along with secretion of substantial amounts of IL-8, I-309 and TARC. Type-2 gamma delta T lymphocytes preferentially expressed the co-stimulatory molecule CD30; conversely, no skewing in chemokine receptor expression was observed. Both polarized populations displayed high levels of CXCR3 in the absence of CCR3, CCR4 and CCR5. Finally, type-1, but not type-2, gamma delta T lymphocytes killed IPP-pulsed U937 cells and displayed elevated perforin content. Overall, our data suggest that type-2 differentiation of gamma delta T lymphocytes profoundly affects both their effector functions and their potential to recruit the appropriate leukocyte subsets to the sites of inflammation.
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- 2002
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8. In vitro stimulation of human tonsillar subepithelial B cells: requirement for interaction with activated T cells.
- Author
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Dono M, Zupo S, Massara R, Ferrini S, Melagrana A, Chiorazzi N, and Ferrarini M
- Subjects
- CD40 Antigens immunology, Cells, Cultured, Epithelium immunology, Humans, Immunoglobulins biosynthesis, Lymphocyte Function-Associated Antigen-1 immunology, B-Lymphocytes immunology, Lymphocyte Activation, Lymphocyte Cooperation, Palatine Tonsil immunology, T-Lymphocytes immunology
- Abstract
Human tonsillar subepithelial B cells, which are a marginal zone-equivalent B cell subset, respond readily to T-independent type 2 antigens, but not to polyclonal B cell activators in vitro. In this study, subepithelial (SE) B cells were induced to proliferate and mature into plasma cells when co-cultured with activated T cells. The response of SE B cells was not observed when co-cultures were carried out in transwell chambers or in the presence of blocking anti-LFA-1 antibodies, demonstrating the need for a close T-B cell interaction. The presence of soluble CD40 also prevented the B cell response in vitro suggesting a pivotal role of CD40-CD40 ligand interactions. The data are discussed in terms of the T cell dependence of marginal zone (MZ) B cell response and the possible existence of various MZ B cell subsets.
- Published
- 2001
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9. Engagement of CD30 shapes the secretion of cytokines by human gamma delta T cells.
- Author
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Biswas P, Rovere P, De Filippi C, Heltai S, Smith C, Dagna L, Poli G, Manfredi AA, and Ferrarini M
- Subjects
- Animals, CD3 Complex physiology, Calcium metabolism, Humans, Mice, Cytokines biosynthesis, Ki-1 Antigen physiology, Receptors, Antigen, T-Cell, gamma-delta analysis, T-Lymphocytes metabolism
- Abstract
CD30 is a member of the TNF receptor superfamily, previously shown to be expressed on Hodgkin's lymphoma cells and on normal activated lymphocytes. We here show that CD30 is highly expressed on recently activated human gamma delta T cells. Elevated surface levels of this molecule persisted in long-term cultures of gamma delta cells, without further cell stimulation. CD30 acted as a co-stimulus in gamma delta T cells by potentiating the intracellular Ca(2+) fluxes induced by CD3 cross-linking. The engagement of CD30 enhanced the expression of several cytokines induced upon CD3 stimulation such as IL-4 and IFN-gamma but not IL-10. The CC chemokines RANTES and macrophage inflammatory protein-1beta were constitutively expressed and not affected by stimulation. The inducible expression of the neutrophil chemoattractant IL-8 was enhanced by CD30 co-stimulation, as well as that of the CC chemokines I-309 and MDC, whereas the secretion of the monocyte chemotactic protein-1 was not detected. Triggering of CD30 may therefore modulate the expression of several cytokines released by gamma delta cells; the expression of its physiologic ligand by APC and neutrophils at the site of infection may contribute to determine the outcome of an immune response.
- Published
- 2000
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10. Identification of HSP-60 as the specific antigen of IgM produced by BRG-lymphoma cells.
- Author
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Musante L, Ulivi M, Cutrona G, Chiorazzi N, Roncella S, Candiano G, and Ferrarini M
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- Amino Acid Sequence, Antibodies, Monoclonal immunology, Apoptosis immunology, Epitope Mapping, Epitopes, B-Lymphocyte immunology, Humans, Molecular Sequence Data, Tumor Cells, Cultured, Antibodies, Neoplasm immunology, Antigens, Neoplasm immunology, Burkitt Lymphoma immunology, Chaperonin 60 immunology, Electrophoresis, Gel, Two-Dimensional methods, Immunoglobulin M immunology, Lymphoma, AIDS-Related immunology
- Abstract
In previous studies we described a patient with Burkitt's lymphoma and AIDS, whose cells recognized a molecule expressed by normal and malignant breast cells. In the present study, we identified this antigen by two-dimensional (2-D) electrophoresis and Western blotting using the antibody produced by lymphoma cells. The antigen so identified consisted of two clusters of spots with a molecular mass (Mr) of 60 and 50 kDa, respectively. Preparative immobilized pH gradient (IPG) was subsequently used to isolate the clusters of spots of higher molecular masses, from which peptide fragments of approximately 10 aa were separated on reverse-phase chromatography and sequenced. This procedure enabled the identification of the antigen recognized by the lymphoma cells as HSP-60. By means of serological analyses it was possible to identify the lower molecular mass cluster of spots as a molecule related to HSP-60. It is hypothesized that this molecule is a membrane form of HSP-60 that differs from HSP-60 in a COOH terminal portion.
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- 1999
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11. Mycobacterium tuberculosis exploits the CD95/CD95 ligand system of gammadelta T cells to cause apoptosis.
- Author
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Manfredi AA, Heltai S, Rovere P, Sciorati C, Paolucci C, Galati G, Rugarli C, Vaiani R, Clementi E, and Ferrarini M
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- Cell Line, Fas Ligand Protein, Humans, T-Lymphocytes microbiology, Apoptosis, Membrane Glycoproteins immunology, Mycobacterium tuberculosis immunology, Receptors, Antigen, T-Cell, gamma-delta immunology, T-Lymphocytes immunology, fas Receptor immunology
- Abstract
Vgamma9/Vdelta2+ T cells specifically recognize Mycobacterium tuberculosis in vitro and are precociously recruited in early mycobacterial lesions. Even if gammadelta T cells are only fortuitously detected in granulomas or bronchoalveolar lavages of patients with active pulmonary tuberculosis, a role in shaping the mature alphabeta T cell response against M. tuberculosis is substantiated. Here we provide a molecular explanation for this paradox: the engagement of the gammadelta TCR by mycobacterial antigens induced the expression of CD95 ligand (CD95L) by chronically activated CD95+/CD95L- gammadelta T lymphocytes. The receptor was functional, as CD95/CD95L interaction triggered the bystander death of CD95+ cells by apoptosis. Cell death was abolished by CD95-blocking antibodies. The transient accumulation at the site of infection of CD95L+ gammadelta lymphocytes, capable of interacting with CD95+ leukocytes attracted by the response towards the pathogen, may determine the characteristics of the ensuing granulomatous disease.
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- 1998
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12. The propensity to apoptosis of centrocytes and centroblasts correlates with elevated levels of intracellular myc protein.
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Cutrona G, Dono M, Pastorino S, Ulivi M, Burgio VL, Zupo S, Roncella S, and Ferrarini M
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- Cell Cycle, Gene Expression, Humans, Palatine Tonsil cytology, RNA, Messenger genetics, Trihexosylceramides analysis, Apoptosis, B-Lymphocytes immunology, Germinal Center cytology, Proto-Oncogene Proteins c-myc genetics
- Abstract
In this study, we investigated the c-myc expression by tonsillar germinal center (GC) B cells using reverse transcriptase-polymerase chain reaction, flow cytometry, Western blot and in situ immunohistochemical methods. The results obtained demonstrate elevated levels of c-myc mRNA and of Myc protein in GC B cells compared to those of the other resting or activated tonsillar B cells. Separation of GC B cells into centroblasts and centrocytes revealed that, while differing in their cell cycle status, surface marker expression and morphology, the two cell types had the same propensity to apoptosis and elevated Myc protein expression, thus reinforcing the notion of a close correlation between these two events. Based upon these observations and other considerations it is proposed that elevation of Myc proteins confers to GC B cells a particular propensity to apoptosis, while the subsequent decision between progression into the cell cycle or programmed cell death is dictated by other signals that are delivered in the GC and perhaps operate at the level of other proto-oncogenes.
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- 1997
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13. Subepithelial B cells in the human palatine tonsil. I. Morphologic, cytochemical and phenotypic characterization.
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Dono M, Burgio VL, Tacchetti C, Favre A, Augliera A, Zupo S, Taborelli G, Chiorazzi N, Grossi CE, and Ferrarini M
- Subjects
- Alkaline Phosphatase metabolism, B-Lymphocytes ultrastructure, CD5 Antigens analysis, Child, Humans, Immunohistochemistry, Immunophenotyping, Palate immunology, B-Lymphocytes immunology, Palatine Tonsil immunology
- Abstract
This study describes the purification of a subset of tonsillar B cells which share phenotypic, morphologic and cytochemical features with subepithelial (SE) B cells. These cells, which represented the 5-10% of the total tonsillar B cells, were found in the Percoll gradient fraction of highest density, together with resting follicular mantle (FM) B cells. The latter B cells, however, expressed surface CD5 and could be removed by an immune rosetting procedure. The remaining small CD5- B cells had a surface phenotype (IgM+, IgD+, CD23-, CD38+/-, CD10-, CD44+) that was different from that of FM (IgM+, IgD+, CD23+, CD39+, CD38-, CD10-, CD44+2) and of germinal center (GC) (CD23-, CD39-, CD38+, CD10+, CD44+/-, IgG+) B cells isolated from the same cell suspensions. Furthermore, the absence of surface activation markers (CD71 and CD69) and of surface IgG allowed us to distinguish small CD5- B cells from activated and memory cells migrating within Percoll fractions of lower density. In situ immunohistochemical studies revealed that B cells with an identical phenotype as that of small CD5- B cells could be detected predominantly in the SE region (lamina propria) of the tonsil, and also within the epithelium lining the cryptae. This area was also comprised of a relatively minor proportion of activated B cells, not found in the small CD5- B cell fraction owing to the separation procedure used. Consistent with the notion that the SE area could be a site of B cell activation was also the presence of activated macrophages and of plasma cells. Thirty to forty percent of small CD5- B cells isolated in suspension were positive for the endogeneous alkaline phosphatase (ALP) activity. In contrast, only a few FM B cells were ALP+, while GC cells were consistently ALP-. In situ studies also demonstrated a prevalent expression of ALP activity by the B cells in the SE area. At the ultrastructural level, small CD5- B cells were clearly different from both FM and GC B cells. They displayed a cytoplasm more extended than that of FM B cells with abundant endosomes and plasma membrane projections, and a speckled pattern of nuclear heterochromatin distribution. When fixed tissue sections were examined, cells with identical ultrastructural features could be demonstrated in the tonsillar lamina propria. Collectively, the above data demonstrate an identity of features between the small CD5- B cells isolated in suspension and SE B cells analyzed in situ. Since tonsillar SE B cells are generally thought to represent the homolog of the extrafollicular B cells (including those of the splenic marginal zone), these studies may provide new opportunities for functional studies on this so far incompletely characterized B cell subset.
- Published
- 1996
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14. Subepithelial B cells in the human palatine tonsil. II. Functional characterization.
- Author
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Dono M, Zupo S, Augliera A, Burgio VL, Massara R, Melagrana A, Costa M, Grossi CE, Chiorazzi N, and Ferrarini M
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- Animals, Antibody Formation, Antigens, CD analysis, Apoptosis, B-Lymphocytes immunology, Humans, Lymphocyte Activation, Lymphocyte Culture Test, Mixed, Mice, Palate immunology, Proto-Oncogene Proteins analysis, Proto-Oncogene Proteins c-bcl-2, Trinitrobenzenes immunology, B-Lymphocytes physiology, Palatine Tonsil immunology
- Abstract
This study investigates the main functional features of subepithelial (SE) B cells and compares them with those of purified germinal center (GC) and follicular mantle (FM) B cells isolated from the same tonsils. Unlike FM B cells, SE B cells failed to produce polyspecific antibodies in vitro; unlike GC B cells, SE B cells expressed high levels of Bcl-2 and failed to undergo spontaneous apoptosis in vitro. The most striking function of SE B cells was their ability to produce IgM antibodies to T cell-independent type-2 (TI-2) (but not to TI-1) antigens (Ag). These antibodies could not be detected when both FM and GC B cells were stimulated with TI-2 Ag in vitro. Moreover, B cells isolated from peripheral blood were unable to mount a response to TI-2 Ag. The latter finding is consistent with the observation that B cells with the phenotypic features of SE B cells were virtually absent in the peripheral blood and emphasizes the notion that SE B cells belong to a subset of non-recirculating B cells. SE B cells were by far superior to FM B cells in mixed lymphocyte reaction (MLR) stimulation of allogeneic T cells in vitro, although they were not as efficient as dendritic cells (DC). In order to stimulate T cells efficiently, SE B cells had to be exposed to anti-mu antibody, a treatment which induced expression of activation markers such as CD80, CD86, CD69 and CD39, usually absent in resting SE B cells. CD80 and CD86 molecules expressed by SE B cells participated in the chain of events required to promote the proliferation of allogeneic T cells as demonstrated by inhibition tests with the appropriate mAb. The expression of CD80 and CD86 by anti-mu-treated SE B cells was not, however, the sole explanation for their good antigen presenting capacities since the exposure of FM B cells to anti-mu antibody also induced expression of these surface structures. Nevertheless, these cells failed to become good MLR stimulators. Collectively, the above data contribute further to the characterization of a distinct subset of tonsillar B cells which resemble, both phenotypically and functionally, the B cells of the splenic marginal zone.
- Published
- 1996
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15. Expression of CD5 and CD38 by human CD5- B cells: requirement for special stimuli.
- Author
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Zupo S, Dono M, Massara R, Taborelli G, Chiorazzi N, and Ferrarini M
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- ADP-ribosyl Cyclase, ADP-ribosyl Cyclase 1, Antibodies, Monoclonal, Antigens, CD physiology, Antigens, Differentiation physiology, CD5 Antigens, Cells, Cultured, Down-Regulation, Epitopes physiology, Flow Cytometry, Fluorescent Antibody Technique, Humans, Interleukin-4 physiology, Lymphocyte Activation, Membrane Glycoproteins, Palatine Tonsil cytology, Tetradecanoylphorbol Acetate, Antigens, CD biosynthesis, Antigens, Differentiation biosynthesis, B-Lymphocyte Subsets immunology
- Abstract
In this study the mode of expression of CD5 by human tonsillar CD5- B cells after stimulation with different agents was investigated. Resting B cells were separated into CD5+ and CD5- cells and the two cell fractions exposed to phorbol 12-myristate 13-acetate (PMA). CD5- B cells expressed CD5 and maximum CD5 expression was achieved after approximately 60 h of culture. Based upon the proportions of cells that express CD5 as well as those of the cells surviving in culture, it was calculated that 15-25% of the total CD5- B cells were induced to express CD5. Unlike CD5- B cells, CD5+ B cells proliferated vigorously in response to PMA as assessed by [3H]thymidine incorporation and cell cycle analysis in vitro. However, the expression of CD5 by CD5- B cells was not related to the selective expansion of some CD5+ B cells left over as contaminant cells since this occurred in the absence of cell proliferation. Upon exposure to PMA, CD5- B cells remained in the G0-G1 phases of the cell cycle and did not express the Ki67 antigen or incorporate [3H]thymidine. Furthermore, mitomycin C treatment of the CD5- B cells did not prevent CD5 expression. Phenotypic studies disclosed that CD5+ B cells but not CD5- B cells expressed CD39. This finding offered the opportunity to carry out an additional control experiment. Separation of the two populations according to the expression of CD39 confirmed the finding obtained by fractionating the cells into CD5+ and CD5- B cells. The cells induced to express CD5 also expressed CD38 that was not detected on resting CD5- B cells. In this respect, the CD5- B cells that converted into CD5+ cells (inducible CD5+ B cells) resembled the cells from the CD5+ B cell fractions that up-regulated CD5 and also expressed CD38 upon exposure to PMA alone. Another example of coordinate expression of these two antigens was the finding that exposure to PMA in the presence of recombinant interleukin-4 (rIL-4) resulted in inhibition of the expression of CD5 and CD38. Although virtually all of the tonsillar CD5- B cells expressed the CD69 activation marker, no cells other than those co-expressing CD5 and CD38 were induced to express CD5 by PMA alone. Resting CD5- B cells failed to express CD5 and/or CD38 when cultured with PMA in the presence of EL4 T cells and IL-4-free T cell supernatants.(ABSTRACT TRUNCATED AT 400 WORDS)
- Published
- 1994
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16. CD38 signaling by agonistic monoclonal antibody prevents apoptosis of human germinal center B cells.
- Author
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Zupo S, Rugari E, Dono M, Taborelli G, Malavasi F, and Ferrarini M
- Subjects
- ADP-ribosyl Cyclase, ADP-ribosyl Cyclase 1, Antibodies, Monoclonal immunology, Blotting, Western, Cells, Cultured, Humans, Lymphocyte Activation, Membrane Glycoproteins, Proto-Oncogene Mas, Proto-Oncogene Proteins biosynthesis, Proto-Oncogene Proteins c-bcl-2, Antigens, CD, Antigens, Differentiation physiology, Apoptosis immunology, B-Lymphocytes cytology, Palatine Tonsil cytology
- Abstract
The present study demonstrates that an agonistic anti-CD38 monoclonal antibody (mAb) (IB4) is capable of preventing apoptosis of human tonsillar germinal center (GC) B cells as measured by either morphological methods on Giemsa-stained cytospin preparations or flow cytometry on propidium iodide-stained cells. Two other anti-CD38 mAb (Leu-17 and OKT10) consistently failed to prevent apoptosis in the same cells, even when tested over a wide range of concentrations. Furthermore, exposure of GC B cells to IB4 mAb up-regulates the bcl-2 proto-oncogene product in a manner similar to that observed with CD40 ligand (CD40L). The ability of IB4 mAb to prevent apoptosis of GC B cells was inferior to that of both anti-CD40 mAb and CD40L. No synergistic or additive effects were observed when IB4 mAb was used together with CD40L. Unlike anti-CD40 mAb or CD40L, IB4 mAb neither induced a proliferation of GC B cells nor increased their proliferative response to anti-CD40, CD40L or recombinant interleukin-4, used alone or in combination. The present results are consistent with the recent findings on either the feature of the CD38 molecules to deliver activation signals and on the mechanisms of selection of B cells that operates in the GC.
- Published
- 1994
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17. Identification of two distinct CD5- B cell subsets from human tonsils with different responses to CD40 monoclonal antibody.
- Author
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Dono M, Zupo S, Masante R, Taborelli G, Chiorazzi N, and Ferrarini M
- Subjects
- Antibodies, Monoclonal immunology, Antigens, CD analysis, Antigens, Differentiation, B-Lymphocyte analysis, Antigens, Differentiation, T-Lymphocyte analysis, CD40 Antigens, Cell Separation, Humans, Immunoglobulin G biosynthesis, Immunoglobulin M biosynthesis, Immunoglobulin mu-Chains immunology, Interleukin-4 pharmacology, Lectins, C-Type, Lymphocyte Activation, Palatine Tonsil cytology, Receptors, Antigen, B-Cell immunology, Antigens, CD immunology, Antigens, Differentiation, B-Lymphocyte immunology, B-Lymphocyte Subsets immunology
- Abstract
This study investigated the response of different CD5- B cell subsets to CD40 monoclonal antibody (mAb) in various combinations with interleukin (IL)-4 or rabbit anti-human mu chain antibody (a-mu-Ab). The different CD5- B cell subsets were isolated from tonsillar B cell suspensions depleted of CD5+ B cells and subsequently fractionated on Percoll density gradients. While resting CD5+ B cells proliferated and produced IgM molecules in response to a-mu-Ab, IL-4 and CD40 mAb as well as to Staphylococcus aureus Cowan strain I (SAC) and IL-2, resting CD5- B cells, which were co-purified in the same 60% Percoll fractions, consistently failed to respond. These cells were, however, activated by the stimuli employed, as demonstrated by their capacity to express the surface activation markers CD69, CD25 and CD71. Resting CD5+ B cells had the typical phenotype of mantle zone B cells (IgM+ IgD+ CD39+ CD38- CD10- CDw75dim), whereas resting CD5- B cells were CD38- CD39- CD10- CDw75 intermediate and expressed surface IgM but relatively little surface IgD and could not be classified as mantle zone or germinal center cells. The finding that purified germinal center cells (CD38+ CD10+ CD39- CDw75bright, IgG+) responded to CD40 mAb and IL-4 and also to SAC plus IL-2 further underlined the differences to resting CD5- B cells. However, some of the data collected suggest possible relationships between CD5- B cells and germinal center cells. The CD5- B cells isolated from the 50% Percoll fraction proliferated in response to a-mu-Ab, CD40 mAb and IL-4 as well as to SAC and IL-2. These cells had the same mantle zone B cell phenotype as the CD5+ B cells, but their capacity to respond to the stimuli in vitro was unrelated to a possible contamination with CD5+ B cells, as documented by the appropriate controls. Furthermore, upon exposure to SAC or phorbol esters, the large majority of CD5- B cells from the 50% Percoll fraction did not express surface CD5 and there was very little if any accumulation of CD5 mRNA. Finally, most of the cycling cells in the stimulated CD5- B cells did not express CD5. The CD5- B cells from the 50% Percoll fraction were comprised of a consistent proportion of cells that expressed surface activation markers.(ABSTRACT TRUNCATED AT 400 WORDS)
- Published
- 1993
- Full Text
- View/download PDF
18. Evidence for differential responsiveness of human CD5+ and CD5- B cell subsets to T cell-independent mitogens.
- Author
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Zupo S, Dono M, Azzoni L, Chiorazzi N, and Ferrarini M
- Subjects
- Antibodies, Anti-Idiotypic, Antigens, Differentiation, B-Lymphocyte biosynthesis, Autoantibodies biosynthesis, B-Lymphocyte Subsets immunology, CD5 Antigens, Cell Cycle, Humans, Immunoglobulin G biosynthesis, Immunoglobulin M biosynthesis, Immunoglobulin mu-Chains physiology, In Vitro Techniques, Lymphocyte Activation immunology, Staphylococcus aureus immunology, Thymidine metabolism, Tritium, Antigens, CD physiology, Antigens, Differentiation physiology, B-Lymphocyte Subsets drug effects, Mitogens pharmacology
- Abstract
Tonsillar resting B cells were separated into CD5+ and CD5- cell subsets and stimulated with the thymus-independent mitogens, Staphylococcus aureus Cowan strain I (SAC) or insolubilized anti-mu monoclonal antibodies (a mu Ab). CD5+ cells incorporated [3H]thymidine more efficiently than unfractionated cells when stimulated with SAC and their response was augmented by the addition of interleukin (IL) 2 to the cultures. CD5+ cells also proliferated in response to a mu Ab provided that IL 2 was present, SAC-, but not a mu Ab-stimulated CD5+ cells produced IgM and IgG molecules when IL 2 was added to the cultures and also secreted autoantibodies with rheumatoid factor activity and sometimes also with anti-single-stranded, but not double-stranded, DNA activity. The efficient response of CD5+ cells was not explained by the fact that they contained cells already activated in vivo. Thus, they did not express the CD23, CD69, CD71 and CD39 activation markers, failed to incorporated [3H]thymidine and to secrete Ig spontaneously or in response to IL 2 and were found to be in a quiescent state by cell cycle flow cytometric analysis. In contrast to CD5+ cells, CD5- cells displayed very little or no [3H]thymidine incorporation in response to SAC or to a mu Ab and their poor responsiveness was not altered by changing either the doses of the stimulants, the timing of the cultures, by co-culturing the cells together with CD5+ cells, or by adding IL 2 or IL 4. Immunofluorescence studies showed that freshly prepared CD5- cells did not have surface activation markers but that they expressed them following SAC stimulation. Thus, unlike that observed for CD5+ cells, SAC seems to be capable of activating CD5- cells but does not appear to be a sufficient stimulus for driving the cells into the subsequent phases of the cell cycle. The above findings, that demonstrate marked differences in the response to CD5+ and CD5- cells to thymus-independent stimuli, may bear relevance for the understanding of the normal clonal expansion of CD5+ cells as well as for the pathogenesis of autoimmune diseases.
- Published
- 1991
- Full Text
- View/download PDF
19. Selective lysis of the autologous tumor by delta TCS1+ gamma/delta+ tumor-infiltrating lymphocytes from human lung carcinomas.
- Author
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Zocchi MR, Ferrarini M, and Rugarli C
- Subjects
- Antibodies, Monoclonal, Antigens, CD analysis, Antigens, CD1, Antigens, Differentiation immunology, Cell Line, Cytotoxicity, Immunologic, Histocompatibility Antigens Class I immunology, Humans, Immunity, Cellular, In Vitro Techniques, Precipitin Tests, Receptors, Antigen, T-Cell classification, Carcinoma immunology, Lung Neoplasms immunology, Lymphocytes, Tumor-Infiltrating immunology, Receptors, Antigen, T-Cell immunology, T-Lymphocyte Subsets immunology
- Abstract
Two distinct non-overlapping populations of TcR1+ (gamma/delta) T cells have been described: the first, bearing the disulfide-linked gamma/delta heterodimer, is predominant in the peripheral blood; the second, expressing the non-disulfide-linked form of TcR1, is mostly confined to epithelial tissues (lung, gut, skin). TcR1+ lymphocytes may be cytotoxic and could be involved in anti-tumor immunity, especially against tumors at epithelial sites. Freshly derived tumor-infiltrating lymphocytes (TIL) obtained from two patients with lung cancer were enriched in CD3+WT31- cells. The percentage of this subset substantially increased upon culture in the presence of interleukin 2. These cells were TcR1+ as demonstrated by immunofluorescence and immunoprecipitation. In one case only 40% of this population reacted with delta TCS1 mAb, that recognizes the non-disulfide-linked form of TcR1, and co-expressed the CD8 antigen. Cultured TcR1+ TIL were able to kill fresh autologous tumor cells, K-562 and, to a lesser extent, some natural killer-resistant cell lines and allogeneic lung tumor cells in 4-h 51Cr-release cytotoxicity assays. The fractionated delta TCS1+ TIL lysed only autologous tumor cells and K-562, whereas the lytic activity against all the other targets was confined to the delta TCS1- subset. Moreover, the autotumor cytotoxicity was inhibited by anti-HLA class I but not by anti-CD1c or anti-LFA-1 mAb, suggesting that killing of the autologous tumor cells and non-major histocompatibility complex-restricted cytotoxicity are mediated by different mechanisms.
- Published
- 1990
- Full Text
- View/download PDF
20. Infection of Epstein-Barr virus-transformed lymphoblastoid B cells by the human immunodeficiency virus: evidence for a persistent and productive infection leading to B cell phenotypic changes.
- Author
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De Rossi A, Roncella S, Calabro ML, D'Andrea E, Pasti M, Panozzo M, Mammano F, Ferrarini M, and Chieco-Bianchi L
- Subjects
- Antigens, Differentiation, B-Lymphocyte analysis, B-Lymphocytes immunology, Cell Line, Genotype, HIV Infections complications, HIV Infections immunology, Humans, Lymphocyte Function-Associated Antigen-1 analysis, Lymphoma, Non-Hodgkin etiology, Phenotype, Proto-Oncogene Mas, Receptors, Fc analysis, Receptors, IgE, B-Lymphocytes microbiology, CD4 Antigens analysis, Cell Transformation, Viral, HIV-1 growth & development, Herpesvirus 4, Human genetics
- Abstract
Four Epstein-Barr virus-positive lymphoblastoid cell lines (LCL) were successfully infected in vitro with immunodeficiency virus type 1 (HIV-1) as demonstrated by reverse transcriptase activity and p24 HIV antigen in culture supernatants, positive cell staining for gag-encoded HIV proteins, presence of viral HIV genome by Southern blot analysis and ulstrastructural observations. In addition, both HIV-1-infected B cells and their supernatants efficiently transactivated the chloramphenicol acetyl transferase reporter gene which is under the control of the HIV-1 long terminal repeat. The LCL cells displayed long-term HIV-1 infection and production, but no cytopathic effects were observed. Cytofluorimetric analysis did not detect membrane CD4 presence in the LCL cells before and after HIV-1 infection; moreover, a minute amount of CD4 mRNA was observed only in one of the LCL. A monoclonal antibody specific for the viral binding site of the CD4 molecule delayed, but did not block, HIV-1 infection of the LCL cells. Following HIV-1 infection, changes in LCL phenotype were observed, consisting of a decrease in CD23- and CD39-positive cells, and a concomitant increase of cells with surface CD10 and Bac-1. Furthermore, HIV-1-infected LCL cells did not grow in tight clumps, as usually observed in uninfected LCL, but as disperse suspensions, and formed more agar colonies than control LCL. However, despite this apparent acquisition of a malignant-like phenotype, c-myc proto-oncogene rearrangement was not detected. The appearance of cells with new characteristics did not seem due to clone selection by HIV-1 infection, since all the LCL conserved their clonotypic pattern of IgH chain rearrangement. The acquisition of malignant-like features by HIV-infected B cells might be clinically significant in terms of the pathogenesis of non-Hodgkin's B cell lymphomas, which occur frequently in AIDS patients.
- Published
- 1990
- Full Text
- View/download PDF
21. Receptors for IgG molecules on human lymphocytes forming spontaneous rosettes with sheep red cells.
- Author
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Ferrarini M, Moretta L, Abrile R, and Durante ML
- Subjects
- Binding Sites, Cell Adhesion, Cell Membrane immunology, Erythrocytes immunology, Humans, Leukemia, Lymphoid immunology, T-Lymphocytes ultrastructure, Immunoglobulin G, T-Lymphocytes immunology
- Abstract
A mixed rosette technique with sheep red blood cells (SRBC) and ox erythrocytes heavily coated with rabbit antibody (EA) was employed to simultaneously identify human peripheral blood T lymphocytes and IgG receptor-bearing cells. The findings of a noticeable proportion of "mixed rosettes" in peripheral blood lymphocytes freshly drawn from normal individuals and of an even higher number of these mixed rosettes in cell suspension kept in culture media supplemented with fetal calf serum provide evidence for the capacity of human T cells to express membrane receptors for antigen-antibody complexes.
- Published
- 1975
- Full Text
- View/download PDF
22. Ultrastructure and cytochemistry of human peripheral blood lymphocytes. Similarities between the cells of the third population and TG lymphocytes.
- Author
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Ferrarini M, Cadoni A, Franzi AT, Ghigliotti C, Leprini A, Zicca A, and Grossi CE
- Subjects
- Histocytochemistry, Humans, Hydrolases, Lymphocytes classification, Monocytes immunology, Receptors, Antigen, B-Cell, Receptors, Fc, Immunoglobulin G, Lymphocytes ultrastructure, Receptors, Immunologic, T-Lymphocytes
- Abstract
The ultrastructural and cytochemical features of human peripheral blood TG cells (T cells with receptors IgG) and of the cells of the so-called third population (non-T, non-B cells with high avidity receptors for IgG) have been investigated and compared. Both TG and third-population cells (TPC) contained acid hydrolases with a paranuclear localization of alpha-naphthyl acid esterase, beta-glucuronidase or acid phosphatase. At the electron microscopy level, TG and TPC were indistinguishable and displayed rough cell surface, indented nuclei, abundant cytoplasm with predominance of the smooth over the rough membranes and peroxidase-negative granules. A large proportion of cells of the TPC could form rosettes with sheep erythrocytes after treatment with neuraminidase. The observed close similarities between TG and TPC may suggest that both cell types belong to a special subset of T cells. However, the alternative hypothesis that both TG and TPC are part of a subset unrelated to T cells, such as a new non-T, non-B cell population, or even of the monocytic-macrophage lineage, is also discussed.
- Published
- 1980
- Full Text
- View/download PDF
23. Lymphocyte membrane receptors in human lymphoid leukemias.
- Author
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Ferrarini M, Tonda GP, Risso A, and Viale G
- Subjects
- Binding Sites, Cell Adhesion, Cell Membrane immunology, Complement System Proteins metabolism, Dermatitis, Exfoliative pathology, Erythrocytes immunology, Humans, Immunoglobulin G metabolism, Leukemia, Lymphoid immunology, Receptors, Antigen, B-Cell analysis, Syndrome, Time Factors, Leukemia, Lymphoid pathology, Lymphocytes immunology
- Abstract
Membrane-bound immunoglobulins, receptors for the Fc fragment of IgG and receptors for the third component of human or murine complement were used as B cell membrane markers to study peripheral blood lymphocytes from twenty-two patients with chronic lymphatic leukemia (CLL), five patients with acute lymphoblastic leukemia (ALL) and one patient with Sézary syndrome. The capacity of human T cells of forming "spontaneous rosettes" with sheep erythrocytes was employed as T cell membrane marker. In nineteen out of twenty-seven CLL or ALL cases tested a larger percentage of cells than that found in normal individuals expressed at least one of the three B cell membrane markers studied. In the patient with Sézary syndrome the percentage of cells forming "spontaneous rosettes" with sheep erythrocytes was larger than the normal, while cells bearing B cells markers were below the normal values.
- Published
- 1975
- Full Text
- View/download PDF
24. Membrane Ig on MPC11 myeloma cells: correlation between the expression of membrane Ig, a receptor for Ig and the process of secretion.
- Author
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Corte G, Risso A, Ferrarini M, and Bargellesi A
- Subjects
- Binding Sites, Antibody, Cell Line, Cycloheximide pharmacology, Immunoglobulins metabolism, Kinetics, Lymph Nodes immunology, Pronase, Protein Binding, Immunoglobulin G metabolism, Myeloma Proteins metabolism, Receptors, Antigen, B-Cell metabolism
- Abstract
The expression of membrane immunoglobulin (mIg) was examined in three cloned MPC11-derived mouse myeloma cell lines. Membrane immunofluorescence studies demonstrated that IgG2b producer cells (P1) had complete IgG molecules, L-chain producer (L1) had only L-chain determinants and nonproducer (NP2) did not have any Ig determinants on the cell surface. An Ig receptor, with characteristics different from B lymphocyte Fc receptor, has been found to be present on secreting cells (P1 or L1), but not on the NP2 cell variant. The data reported in the present paper indicate that the expression of mIg and of the Ig receptor molecule is clearly correlated with the process of secretion. In the light of previous data reported on Ig secretion, a model is proposed which correlates the process of secretion with the expression not only of mIg, but also of the receptor for Ig.
- Published
- 1976
- Full Text
- View/download PDF
25. Human T cell receptor for IgM: specificity for the pentameric Fc fragment.
- Author
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Ferrarini M, Moretta L, Mingari MC, Tonda P, and Pernis B
- Subjects
- Binding Sites, Humans, Immunoglobulin Fc Fragments, Immunoglobulin M, T-Lymphocytes immunology
- Abstract
The specificity of the IgM receptor expressed by human T cells cultured in IgM-free media has been investigated. IgM receptors have been detected using a rosette system with ox erythrocytes coated with rabbit antibody (EA(IgM)), and the inhibitory capacity of different IgM fragments in the rosette system has been tested. It was found that F(c)5mu but not F(ab')2mu, nor monomeric IgM (8 S IgM) inhibited EA (IgM) rosette-forming cells. This indicates that the receptor present on the surface of T cells has affinity for a structure located in the Fc portion of the pentameric IgM.
- Published
- 1976
- Full Text
- View/download PDF
26. Human T cell lines with antigen specificity and helper activity.
- Author
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Lanzavecchia A, Ferrarini M, and Celada F
- Subjects
- Antibody-Producing Cells immunology, Antigens, Surface immunology, B-Lymphocytes cytology, B-Lymphocytes immunology, Cell Line, Humans, Immunoglobulin M biosynthesis, Lymphocyte Activation, Plasma Cells cytology, T-Lymphocytes cytology, Tetanus Toxoid pharmacology, Epitopes, T-Lymphocytes immunology, T-Lymphocytes, Helper-Inducer metabolism
- Abstract
Human T blasts, obtained by stimulation of peripheral blood mononuclear cells (PBM) with tetanus toxoid, diphtheria toxoid or Candida albicans, were expanded in long-term culture using alternate periods of antigen restimulation and growth in media containing interleukin 2. The cells gave a proliferative response only to the antigen originally used for stimulation. Such as response was strictly dependent upon the presence of autologous but not of allogeneic mitomycin C-treated mononuclear cells. When added to autologous PBM depleted of E-rosetting cells together with the specific antigen, the T blasts induced a polyclonal proliferation and differentiation of B cells. Allogeneic B cells were activated by antigen-stimulated T blasts only in the presence of irradiated mononuclear cells autologous to the responding T blasts. The above responses seemed not to be regulated solely by the release of soluble factors; apparently cell to cell interactions had to take place to obtain an efficient B cell activation.
- Published
- 1982
- Full Text
- View/download PDF
27. CD8+CD11b+ peripheral blood T lymphocytes contain lymphokine-activated killer cell precursors.
- Author
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Dianzani U, Zarcone D, Pistoia V, Grossi CE, Pileri A, Massaia M, and Ferrarini M
- Subjects
- Acid Phosphatase metabolism, Antibodies, Monoclonal, CD8 Antigens, Cell Differentiation, Cells, Cultured, Clone Cells, Cytotoxicity, Immunologic, Humans, In Vitro Techniques, Interleukin-2 pharmacology, Lymphocyte Activation, Macrophage-1 Antigen, Microscopy, Electron, Phytohemagglutinins pharmacology, T-Lymphocytes classification, T-Lymphocytes cytology, Antigens, Differentiation analysis, Antigens, Differentiation, T-Lymphocyte analysis, Killer Cells, Natural immunology, T-Lymphocytes immunology
- Abstract
CD3+CD8+ cells, purified from peripheral blood T cells by depletion of CD4+ lymphocytes, were tested for their cytotoxic activity against K-562 or HL-60 cells. Freshly prepared cells had no cytotoxic function, but upon exposure to recombinant interleukin 2 (rIL2) in vitro they acquired a lymphokine-activated killer (LAK) activity. Fractionation of CD3+CD8+ cells into CD11b+ and CD11b- cells demonstrated that all the cells with the potential of developing LAK cell functions were within the CD8+CD11b+ subset. These cells lacked natural killer cell markers such as CD16 or NKH1, but a proportion of them stained for Leu-7. Furthermore, they expressed the alpha/beta chains, but not the gamma/delta chains, of the T cell receptor, as could be determined by staining with the appropriate monoclonal antibodies. CD8+CD11b+ cells had a large granular lymphocyte morphology, as shown by both cytochemical and electron microscopy analyses. They proliferated in response to IL2 in vitro and developed cytotoxic functions against a number of natural killer resistant targets. Their response to phytohemagglutinin or pokeweed mitogen was very weak or absent. By contrast, CD8+CD11b- cells failed to generate LAK cells in response to rIL2, did not show a large granular lymphocyte morphology, but responded vigorously to phytohemagglutinin or pokeweed mitogen. Purified CD8+CD11b+ cells were cloned by limiting dilution in the presence of rIL2. The observed cloning efficiency of 19 +/- 0.3% indicates that a fraction of the cells only could respond to IL2. Furthermore, only 50% of the clones obtained had a LAK cell function. These data show that CD8+CD11b+ cells represent a heterogeneous cell population. Nevertheless this cell subset probably represents the major source of LAK cell progenitors within the circulating T cells.
- Published
- 1989
- Full Text
- View/download PDF
28. A study of immunoglobulin classes present on the membrane and in the cytoplasm of human tonsil plasma cells.
- Author
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Ferrarini M, Viale G, Risso A, and Pernis B
- Subjects
- Fluorescent Antibody Technique, Humans, Immunoglobulin A, Immunoglobulin D, Immunoglobulin G, Immunoglobulin Light Chains, Immunoglobulin M, Receptors, Antigen, B-Cell analysis, Cytoplasm immunology, Immunoglobulins, Palatine Tonsil immunology, Plasma Cells immunology
- Abstract
The problem of whether immunoglobulin (Ig)-containing plasma cells expressed membrane Ig has been investigated using cells from human tonsils. In tonsils, IgG-containing cells are predominant, but a certain number of IgM, IgA and IgD-containing cells are also present. By using a double staining immunofluorescent technique for the simultaneous detection of membrane and intracytoplasmic Ig, it has been possible to ascertain that the large majority of IgA, IgM and IgD-containing cells had membrane immunoglobulin (mIg) of a class coincident with that of intracytoplasmic Ig. In addition a noticeable proportion of IgM-containing cells expressed membrane IgD, thus indicating that a certain number of these cells bore both membrane IgM and IgD. About 60% of IgG-containing cells had membrane IgG, while the remaining cells did not express mIg. Furthermore the surface staining of these cells was generally fainter than that of the cells containing other Ig classes. Experiments on the surface light chain type expressed by the single Ig-containing cells (IgCC) as compared to that found in the cytoplasm have shown that in the large majority of IgCC the light chain type of mIg coincided with that of intracytoplasmic Ig. Discordant light chain types of membrane and cytoplasmic Ig were found on about 12% of IgCC only. These values can be taken as a measure of how many IgCC had passively acquired mIg.
- Published
- 1976
- Full Text
- View/download PDF
29. Expression of a receptor for IgM by human T cells in vitro.
- Author
-
Moretta L, Ferrarini M, Durante ML, and Mingari MC
- Subjects
- Antigen-Antibody Complex, B-Lymphocytes immunology, Binding Sites, Blood, Cell Membrane immunology, Cells, Cultured, Culture Media, Humans, Immunoglobulin G metabolism, Membrane Proteins metabolism, Pronase, Immunoglobulin M metabolism, T-Lymphocytes immunology
- Abstract
The capacity of human peripheral blood lymphocytes (PBL) to bind antigen-IgG antibody (EA(IgG)) and antigen-IgM antibody (EA(IgM)) complexes was investigated using a rosette technique with ox erythrocytes coated with rabbit IgG or IgM antibody. It was found that while EA(IgG)-rosette-forming cells (RFC) were detected on PBL freshly drawn from normal individuals, EA(IgM)-RFC were present only in suspensions kept in culture for 24 h in mediá supplemented with sera containing very low or no amounts of IgM. Experiments of simultaneous detection of EA(IgG)-RFC or EA(IgM)-RFC and other membrane markers for human T or B cells together with experiments on purified T or B cell populations indicated that EA(IgG)-RFC were formed by both T and B cells, while T cells only were capable of EA(IgM) rosette formation. The specificity of the receptors for IgG and IgM was determined by studying the inhibitory capacity of purified human IgM and IgG in the rosette assay. The receptor for IgG was inhibited by IgG and not by IgM, while the reverse was true for the receptor for IgM.
- Published
- 1975
- Full Text
- View/download PDF
30. Membrane Ig on human lymphocytes: rate of turnover of IgD and IgM on the surface of human tonsil cells.
- Author
-
Ferrarini M, Corte G, Viale G, Durante ML, and Bargellesi A
- Subjects
- Cells, Cultured, Humans, Iodine Radioisotopes, Kinetics, Pronase, Immunoglobulin D metabolism, Immunoglobulin M metabolism, Lymphocytes immunology, Palatine Tonsil immunology, Receptors, Antigen, B-Cell metabolism
- Abstract
The turnover of IgM and IgD molecules present on the membrane of human tonsil cells has been studied using immunofluorescence and peroxidase-catalyzed membrane radioiodination. With the first of the two techniques cells were treated with pronase to remove membrane immunoglobulin (mIg), placed in culture and stained at intervals to check the reappearance of membrane IgD and IgM on the cell membrane. These experiments showed that membrane IgD (in contrast to membrane IgM) are extremely susceptible to proteolysis. Furthermore, cells treated with a concentration of pronase found to be optimal to remove membrane IgM failed to re-express membrane IgD in vitro. The large majority of tonsil lymphocytes has both membrane IgM and IgD. Due to the different behavior of reappearance of the two membrane molecules after treatment with pronase, it was not possible to obtain the simultaneous re-expression of membrane IgM and IgD by the cells "stripped" with pronase. However, the two molecules were re-expressed in vitro by the cells treated with different pronase concentrations with a similar timing, i.e. 50% or more of the cells re-expressed membrane IgD and IgM after 8 h in culture. 131I-radioiodinated membrane IgD and IgM were also released from the cell surface with a similar timing, the half-life of permanence on the cell membrane being about 4 h for both molecules. These findings thus indicate that IgM and IgD molecules have a similar turnover and that a cell is capable of placing two different Ig molecules at a time on its surface.
- Published
- 1976
- Full Text
- View/download PDF
31. Presence of serum IgD and IgD-containing plasma cells in the mouse.
- Author
-
Bargellesi A, Corte G, Cosulich E, and Ferrarini M
- Subjects
- Animals, Electrophoresis, Polyacrylamide Gel, Fluorescent Antibody Technique, Immune Sera, Immunoglobulin Light Chains, Immunoglobulin delta-Chains, Mice, Mice, Inbred BALB C, Molecular Weight, Peyer's Patches immunology, Spleen immunology, Immunoglobulin D, Plasma Cells immunology
- Abstract
A new technique which makes use of anti-delta-specific allo-or heteroantisera coupled to Sepharose 4 B has been employed to investigate the presence of IgD molecules in the sera of three mouse strains. The sera were reacted with the insoluble immunosorbents, and the eluted material was labeled with 125I prior to antigenic or sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. This material reacted with anti-delta or anti-L chain reagents, but not with other anti-class-specific antisera and, after reduction, was resolved in three discrete bands in SDS-PAGE. The fastest of these bands comigrated with L chain, whereas the other two moved identically to mouse lymphocyte surface delta chains. Immunofluorescence studies on fixed smears of spleen and Peyer's patches' cells revealed the presence of a small but consistent number IgD-containing plasma cells.
- Published
- 1979
- Full Text
- View/download PDF
32. Lymphocyte membrane immunoglobulins: similarities between human IgD and mouse IgD-like molecules.
- Author
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Sitia R, Corte G, Ferrarini M, and Bargellesi A
- Subjects
- Animals, Electrophoresis, Polyacrylamide Gel, Fluorescent Antibody Technique, Humans, Iodine Radioisotopes, Mice, Mice, Inbred BALB C, Molecular Conformation, Molecular Weight, Pronase pharmacology, Immunoglobulin D metabolism, Lymphocytes immunology, Receptors, Antigen, B-Cell
- Published
- 1977
- Full Text
- View/download PDF
33. Cytophilic antibody: correlation of its distribution with activation of basophils and macrophages.
- Author
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Ferrarini M, Munro A, and Wilson AB
- Subjects
- Animals, Binding Sites, Antibody, Epitopes, Female, Fluorescent Antibody Technique, Guinea Pigs, Immune Sera, Immunity, Cellular, Immunoglobulin Fab Fragments, Immunoglobulin G, Immunologic Techniques, Lymphocytes immunology, Male, Microscopy, Phase-Contrast, Peritoneal Cavity immunology, Rabbits immunology, Antibodies, Basophils immunology, Macrophages immunology
- Published
- 1973
- Full Text
- View/download PDF
34. The detection of theta antigen on cells by immunofluorescence: failure to detect theta antigen on rosette-forming cells.
- Author
-
Russell SM, Ferrarini M, Munro A, and Lachmann PJ
- Subjects
- Animals, Bone Marrow immunology, Bone Marrow Cells, Complement System Proteins, Erythrocytes immunology, Immune Sera, Mice, Mice, Inbred BALB C, Mice, Inbred CBA, Sheep immunology, Spleen cytology, Fluorescent Antibody Technique, Histocompatibility Antigens analysis, Immune Adherence Reaction, T-Lymphocytes immunology
- Published
- 1972
- Full Text
- View/download PDF
35. Allotypic determinants on the surface of rosette-forming cells in the rabbit.
- Author
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Ferrarini M, Kent SP, Munro A, Kelus AS, Catty D, and Coombs RR
- Subjects
- Animals, Antibodies, Anti-Idiotypic, Erythrocytes immunology, Female, Immune Sera, Immunization, Immunoglobulin G, Male, Rabbits, Sheep immunology, Epitopes, Immune Adherence Reaction, Isoantigens
- Published
- 1973
- Full Text
- View/download PDF
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