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S Ferrone

Publications and source records attributed to S Ferrone.

At least 271 records · Page 15Linked to original sources
PubMed1985

Distribution of a cross-species melanoma-associated antigen in normal and neoplastic human tissues.

In previous studies the monoclonal antibody (MoAb) M2590 elicited in C57/BL6 mice with the syngeneic melanoma cell line B16 has been shown to recognize a 31K glycoprotein expressed by human melanoma cell lines. The present study has shown that the MoAb M2590 cross-reacts with surgically removed benign and malignant lesions of melanocyte origin. The reactivity pattern of the MoAb M2590 with these lesions is different from that of the anti-high-molecular weight-melanoma associated antigen (HMW-MAA) MoAb 225.28S, of the anti-115K MAA MoAb 345.134S, and of the anti-100K MAA MoAb 376.96S, which were elicited with human melanoma cell lines. In particular, the MoAb M2590 reacts with blue nevi. The MoAb M2590-defined MAA, like other types of MAA, is heterogeneous in lesions removed from different patients, in autologous lesions removed from different anatomic sites, and in cells within a lesion. The distribution of the MoAb M2590-defined MAA in normal tissues and in tumors of nonmelanocyte origin is broader than that of the HMW-MAA, but is similar to that of the 115K MAA and of the 100K MAA. The results of this investigation suggest that immunization with xenogeneic melanoma cells may broaden the range of specificity of antihuman MAA MoAbs and provide information about the phylogenetic evolution of MAAs.

Adult↗
PubMed1985

The monoclonal antibody AC1.59 defines a new polymorphic determinant on HLA-DR molecules.

2-dimensional gel electrophoresis analysis and sequential immunoprecipitation studies have shown that the monoclonal antibody (MoAb) AC1.59 recognizes a gene product of the HLA-DR locus. The determinant defined by the MoAb AC-1.59 has an unusual distribution on HLA-DR allospecificities, since it is expressed by HLA-DR1, DRw8, DRw9 antigens and by subtypes of HLA-DRw6 and HLA-DR4 antigens. In the latter allospecificity the expression of the MoAb AC1.59 defined determinant correlates with the cellularly defined HLA-Dw4 alloantigen. The present investigation provides evidence of serological heterogeneity within HLA-DR alloantigens. Furthermore these results suggest that monoclonal antibodies can sharpen the discriminatory power of serological assays to dissect the heterogeneity and complexity of HLA-DR alloantigens.

Antibodies, Monoclonal↗
PubMed1985

Regulatory role of a monomorphic determinant of HLA Class I antigens in T cell proliferation.

The role of HLA Class I antigens in T cell proliferation was investigated by using the anti-HLA Class I monoclonal antibodies (MoAb) CR10-215, CR10-325, and CR11-115. MoAb CR10-215 and CR11-115 recognize the same (or spatially close) monomorphic determinant, which is distinct and spatially distant from that reacting with MoAb CR10-325. Addition of MoAb CR10-215 and CR11-115 to cultures of peripheral blood mononuclear cells stimulated with MoAb OKT3, MoAb Pan T2, PHA, or PPD inhibited cell proliferation. The blocking is specific in that the anti-HLA Class I MoAb CR10-325 and the Pan T MoAb Pan T1 had no effect on the proliferation. The inhibitory activity of MoAb CR10-215 and CR11-115 does not reflect i) toxic effects, ii) induction of suppressor cells and factors, iii) blocking of the binding of mitogens to lymphocytes, iv) inhibition of the production of interleukin 1 (IL 1) and interleukin 2 (IL 2), or v) function of IL 2 receptor. Anti-HLA Class I MoAb were able to inhibit the proliferation of purified, Tac-, T cells. The inhibited cells did not express Tac antigen, as assayed by direct immunofluorescence, with MoAb anti-Tac, but released a normal amount of IL 2 in culture medium. These results indicate that monomorphic determinants of the HLA Class I complex are involved in the regulation of T cell proliferation. The effect appears to occur at the level of IL 2 receptor expression.

Antibodies, Monoclonal↗
PubMed1985

Analysis of the interaction between a human high molecular weight melanoma-associated antigen and the monoclonal antibodies to three distinct antigenic determinants.

The restricted tissue distribution and the limited heterogeneity that appear in melanoma lesions of the high M.W. melanoma-associated antigen (HMW-MAA) suggest that this antigen may be an appropriate marker for radioimaging, and a useful target for immunotherapy in patients with melanoma. Therefore, in this study we analyzed other characteristics that are important in the selection of reagents for radioimaging and immunotherapy purposes. The affinities of the monoclonal antibodies (MoAB) 149.53, 225.28S, and 763.74T to distinct determinants of the HMW-MAA were found to be at least 1 X 10(8) mol/L. Furthermore, the effects of the concentrations of unlabeled MoAb on the dissociation rates suggest that the binding of MoAb 149.53 and 225.28S to melanoma cells (Colo 38) is preferentially bivalent, whereas that of MoAb 763.74T is preferentially univalent. These results suggest that the latter MoAb is the reagent of choice for assays that make use of soluble HMW-MAA, whereas the former two are the reagents of choice for assays with membrane-bound HMW-MAA, such as imaging with radiolabeled MoAb. The density of the HMW-MAA on cultured Colo 38 melanoma cells appears to be in the range of approximately 5 X 10(6) molecules/cell. The HMW-MAA was not susceptible to MoAb-mediated modulation under a variety of experimental conditions that included various concentrations of modulating MoAb, different incubation times, the use of an anti-mouse Ig antiserum, and the relaxation of equilibrium by diluting cells in MoAb-free medium. These results indicate that the HMW-MAA and the available corresponding MoAb meet the criteria to be reagents for radioimaging and immunotherapy in patients with melanoma.

Animals↗
PubMed1985

Heterogeneous expression of melanoma-associated antigens and HLA antigens by primary and multiple metastatic lesions removed from patients with melanoma.

Indirect immunofluorescence staining with a large battery of monoclonal antibodies of primary and autologous metastatic lesions removed from seven patients with melanoma has detected heterogeneity in the expression of various types of melanoma-associated antigens (MAAs), of distinct determinants of the high molecular weight melanoma-associated antigen (HMW-MAA), of the two subunits of Class I HLA antigens, and of the gene products of the HLA-D region. Among the 10 MAAs tested, the HMW-MAA had the highest frequency and the Mr 87,000 MAA the lowest. Furthermore, the HMW-MAA displayed the lowest heterogeneity. These findings, in conjunction with the restricted tissue distribution of the HMW-MAA, its lack of susceptibility to antibody-mediated modulation, and the high affinity of the available anti-HMW-MAA monoclonal antibodies, indicate that this antigen may be a useful marker for radioimaging and immunotherapy in patients with melanoma. The common acute lymphoblastic leukemia antigen was detected only in five lesions. Class I HLA antigens were detected in a larger number of lesions than HLA-DR antigens, which had a significantly higher frequency than HLA-DQ antigens. The degree of antigenic heterogeneity did not appear to correlate with the histopathological features of the lesions and/or with the clinical course of the disease. The results of the present study indicate that immunodiagnostic and immunotherapeutic approaches to melanoma should rely on the use of combinations of monoclonal antibodies to distinct MAAs.

Animals↗
PubMed1985

Antigenic profile and functional characterization of human peritoneal macrophages.

The antigenic profile and the functional properties of human peritoneal macrophages have been analyzed by using a panel of monoclonal antibodies (MoAb) and functional assays. All peritoneal macrophages were stained by the anti-class I HLA MoAb Q6/64. Between 40 and 100% of the cells were stained by the anti-HLA-DR + DP MoAb Q2/80, Q5/6, and Q5/13; approximately 80% of the cells were stained by the anti-HLA-DQ MoAb BT3.4, and about 95% were stained by the anti-macrophage MoAb OKM1. Peritoneal macrophages were not stained by the anti-dendritic cells MoAb Ki-M4 or by MoAb to T cell subsets, although all of the MoAb were reactive with the appropriate substrates. More than 60% of the cells expressed Fc receptors and C3 receptors, and displayed phagocytic activity. Peritoneal macrophages were effective in stimulating autologous and allogeneic lymphocytes and in presenting soluble antigens to T cells. These reactions were blocked by the anti-HLA-DR + DP MoAb Q5/13, but were not affected by the anti-dendritic cells MoAb Ki-M4 or by the anti-class I HLA MoAb Q6/64. These results suggest that human peritoneal macrophage preparations, without detectable contamination with dendritic cells, can induce proliferation of autologous and allogeneic T cells, and that class II HLA antigens play a significant role in these phenomena.

Antigen-Presenting Cells↗
PubMed1985

Uptake of methotrexate linked to polyclonal and monoclonal antimelanoma antibodies by a human melanoma cell line.

[3H] Methotrexate [( 3H]MTX) was covalently linked to monoclonal antibody (MoAb) 225.28S against human melanoma, to a rabbit anti-human melanoma IgG absorbed either with human red blood cells (AHMGR) or with red blood cells and a variety of normal human tissues (AHMGR + T), or to normal rabbit IgG (NRG). Human melanoma M21 cells were incubated at 0 degrees C or 37 degrees C with 10 microM free MTX or 10 microM MTX linked to one of the above carriers. The order of net uptake of MTX during 6 hours was MTX-MoAb 225.28S greater than MTX-AHMGR greater than MTX-AHMGR + T greater than MTX-NRG greater than or equal to MTX. This order of uptake by the three antibody conjugates corresponded to the amount of conjugate bound at equilibrium at 0 degrees C and to the immunofluorescence titers. Binding sites for MoAb 225.28S were more efficient for internalization of MTX than were those for the two polyclonal antibody preparations. When M21 cells preloaded with MTX by incubation at a drug concentration of 1.0 or 10 microM were incubated in drug-free medium, the amount of cell-associated MTX rapidly declined to 1.8 pmol/mg protein, i.e., the level of intracellular dihydrofolate reductase (DHFR). However, when cells preloaded to a drug content of 112 pmol/mg protein by incubation with 10 microM MTX linked to AHMGR were transferred to conjugate-free medium, 65 pmol MTX/mg remained cell associated after 12 hours. The efflux was inhibited by chloroquine. Both the efflux medium and M21 cells after a 9.5-hour incubation period had MTX-containing catabolic fragments that inhibited DHFR.

Absorption↗
PubMed1985

Appearance of Ia antigens on T lymphocytes stimulated by streptococcal mitogen as an in vitro model of autoimmunity.

Streptococcal mitogen (SM) is an extracellular product of group A streptococci, which is nonspecifically mitogenic for both B and T lymphocytes. The mitogenic activity of SM is resistant to digestion with trypsin, to heating at 100 degrees C for 5 min and to treatment with dithiothreitol. The proliferative response of lymphocytes from patients with a history of rheumatic fever is similar to that of lymphocytes from healthy donors when stimulated with optimal concentrations of SM, but is significantly reduced when low doses of SM are used. T lymphocytes stimulated with SM acquire la antigens and the ability to stimulate allogeneic and autologous lymphocytes in mixed lymphocyte reactions. An involvement of la antigens in these reactions is indicated by the specific block by monoclonal antibodies. T lymphocytes may acquire la antigens following exposure to an appropriate antigen. If this occurs following a streptococcal infection, these in vitro findings suggest that la bearing T cells may play a role in the immunopathological events which can follow a streptococcal infection. The system described may be a useful in vitro model to analyse the immunopathological events which can follow a streptococcal infection and to develop therapeutic approaches to autoimmune conditions.

Autoimmune Diseases↗
PubMed1985

In vivo and in vitro immunomodulatory ability of streptococcal mitogen: effects on adjuvant disease in the rat; on homograft rejection in the mouse; and on the appearance of Ia antigens on human T lymphocytes.

Streptococcal mitogen (SM) is an extracellular product of group A streptococci, nonspecifically mitogenic for both B and T lymphocytes. The mitogenic activity of SM is resistant to digestion with trypsin, to heating at 100 degrees C for 5 min and to treatment with dithiothrietol. The proliferative response of lymphocytes from patients with a history of rheumatic fever is similar to that of lymphocytes from healthy donors when stimulated with optimal concentrations of SM, but is significantly reduced when low doses of SM are used. Rats were treated with s.c. injections of SM for 21 days at various times from the injection with adjuvant: both primary and secondary reactions in the joints were depressed, more so with pretreatment and early treatment. A similar pattern of response was observed in the survival of A tail skin grafts on the flank of CBA mice: a strain combination with strong H-2 incompatibility. Human T lymphocytes stimulated with SM acquired Ia antigens and the ability to stimulate allogeneic and autologous lymphocytes in mixed lymphocyte reactions. An involvement of Ia antigens in these reactions was indicated by the specific block by monoclonal antibodies to human Ia antigens. T lymphocytes may acquire Ia antigens following exposure to an appropriate antigen. If this occurs following a streptococcal infection, our in vitro findings suggest that Ia bearing T cells may play a role in the immunopathological events which can follow a streptococcal infection. This in vitro system may be a useful model to analyse these immunopathological events and to develop therapeutic approaches in autoimmune conditions. The in vitro findings, together with the observed suppressive effects on the in vivo models described, indicate that SM is a valuable means for study and manipulation of a variety of immune responses.

Adjuvants, Immunologic↗
PubMed1985

Suppression of human melanoma growth in nude mice injected with anti high-molecular-weight melanoma-associated antigen monoclonal antibody 225.28S conjugated to purothionin.

The cytotoxic agent purothionin purified from barley was covalently conjugated to the anti-human high-molecular-weight melanoma-associated antigen (HMW-MAA) monoclonal antibody (MoAb) 225.28S by utilizing water-soluble carbodiimide. Injection of the conjugate (0.1 mg/injection) significantly increased the life span of nude mice injected with ascitic-form human melanoma cells (Colo 38), and caused 40% inhibition on day 25 of the growth of solid-form human melanoma cells in nude mice. The effect is specific and is markedly influenced by the site of growth of tumors and by the schedule of administration of the conjugate.

Animals↗
PubMed1985

Immunochemical analysis of the modulation of human melanoma-associated antigens by DNA recombinant immune interferon.

By utilizing the human melanoma cell line Colo 38, a panel of monoclonal antibodies, and a combination of serologic and immunochemical assays, the effect of recombinant immune interferon (IFN-gamma) on the synthesis, expression, and shedding of a cytoplasmic melanoma-associated antigen (MAA) and of the membrane-bound high m.w. MAA (HMW-MAA), 115K MAA, and 100K MAA has been investigated. IFN-gamma increased the synthesis and shedding of the cytoplasmic MAA, but reduced the synthesis and cell surface expression of the HMW-MAA and of the 100K MAA. The cell surface expression of the 115K MAA on IFN-gamma-treated melanoma cells was reduced, although its synthesis was not markedly changed. The effects were dose-dependent and were related to the incubation time of cells with IFN-gamma. Among the three membrane-bound MAA analyzed, the 100K MAA was the most susceptible to modulation by IFN-gamma. The effects of IFN-gamma preparations are not mediated by contaminants in IFN-gamma preparations because removal of IFN-gamma by affinity chromatography on anti-IFN-gamma monoclonal antibodies abolished its modulating activity. The effects of IFN-gamma on the cytoplasmic MAA are similar to those of leukocyte and fibroblast interferons, whereas those on the membrane-bound MAA are significantly different. The potential implications of the marked changes in the antigenic profile of melanoma cells treated with IFN-gamma are discussed in view of the changes in the immunogenicity of IFN-gamma-treated melanoma cells.

Animals↗
PubMed1985

Differential modulation by recombinant immune interferon of the expression and shedding of HLA antigens and melanoma associated antigens by a melanoma cell line resistant to the antiproliferative activity of immune interferon.

By culture of human melanoma Colo 38 cells in the presence of increasing concentrations of recombinant immune interferon (IFN-gamma), the clone RZ gamma-4G.1 resistant to the antiproliferative action of IFN-gamma (3 X 10(4) units/ml) was isolated. This clone was cultured for 6 weeks in the absence of IFN-gamma and was subsequently treated with increasing concentrations of IFN-gamma. Contrary to the response of its parental cell line, treatment of this clone with IFN-gamma did not significantly alter the rate of protein or DNA synthesis and did not markedly modulate the cell surface expression of HLA Class I antigens, of the high molecular weight melanoma associated antigen, and of a Mr 100,000 melanoma associated antigen. IFN-gamma caused an increase in the cell surface expression and in the shedding of HLA Class II antigens from IFN-gamma resistant cells. Four proteins with molecular weights of 32,000, 38,000, 46,000, and 50,000 were induced by IFN-gamma in the parental melanoma cells but not in the resistant clone. Both cell lines bound equivalent amounts of 125I-IFN-gamma to their surface, indicating that the lack of specific surface receptors was not the cause of insensitivity to IFN-gamma. These results indicate that at the cellular level IFN-gamma modulates the expression and shedding of HLA Class II antigens through different mechanisms to those responsible for the antiproliferative action, modulation of the cell surface expression of melanoma associated antigen and of HLA Class I antigens, and induction of new proteins in cultured melanoma cells. Since the success of therapy of malignant diseases with IFN-gamma depends on the extent of resistance of individual tumor cells, the present study may provide a better understanding of the biology of IFN-gamma insensitive tumor cells and particularly the malignant melanoma.

Animals↗
PubMed1985

Immunoreactivity and biodistribution of indium-111-labeled monoclonal antibody to a human high molecular weight-melanoma associated antigen.

The anti-human, high molecular weight-melanoma associated antigen (HMW-MAA) MoAb 225.28S was chelated with 111In and then tested for its in vitro reactivity with cultured human melanoma cells and for its biodistribution in human melanoma bearing nude mice. In vitro studies showed that the radiolabeled antibody reacted specifically with cultured melanoma cells. However, binding of DTPA to the monoclonal antibody reduced its titer with cultured melanoma cells from 1:1024 to 1:512. Further labeling of the DTPA-antibody conjugate with 111In caused an additional reduction of its titer to 1:128. Injection of the radiolabeled monoclonal antibody into nude mice resulted in the accumulation of significantly (p less than 0.001) higher radioactivity in melanoma tissue than in nude mice injected with either [111In] chloride or 111In-labeled antibody to human acid phosphatase. The specificity of the distribution of the radiolabeled antibody in nude mice also was indicated by its poor localization in lesions other than melanoma (e.g., human prostate carcinoma and chronic abscess). The localization of antibody in liver and kidney was also high, although lower than that achieved in tumor. These results indicate that 111In-labeled monoclonal antibodies to human tumor associated antigens may be useful for localizing malignant lesions. However, there is a need to improve labeling and/or purification of antibody in order to decrease renal and hepatic activity.

Animals↗
PubMed1984

Human T cells in cord blood: abnormalities in IA antigens induction by phytohemagglutinin and in autologous mixed lymphocyte reactions.

About 25% of human T cells isolated from cord blood acquired la antigens, following stimulation with phytohemagglutinin (PHA) for 72 hr. This percentage is markedly lower than that found in PHA-activated T-cell populations (PHA-T cells) isolated from peripheral blood of adults. The low expression of la antigens by human T cells from cord blood does not reflect abnormalities in the sensitivity to PHA stimulation and/or in the kinetics of induction of la antigens. PHA-T cells from cord blood display a low stimulatory activity in autologous mixed lymphocyte reactions (MLR). The defect does not reflect a nonspecific abnormality in the stimulatory activity of PHA-T cells from cord blood, since the latter do not differ from PHA-T cells from adults in their ability to stimulate allogeneic T cells from adults. Furthermore the defect does not reflect a nonspecific abnormality in the proliferative response of T cells from cord blood, since the latter display a normal proliferative response to PHA-T cells from adults. The defect in the proliferative response is not restricted to the autologous MLR with PHA-T cells, since it was found also in autologous MLR with non-T cells as stimulators. Correlation of the temporal evolution of the abnormalities of human T cells with the maturation of the immune system may contribute to our understanding of the role of la antigens in cell-cell interactions and of the biological significance of abnormalities of autologous MLR.

Cells, Cultured↗
PubMed1984

Reactivity patterns with HLA-A2 variants indicate lack of identity between determinants defined by monoclonal antibodies and cytotoxic-T-cell clones.

The anti-HLA-A2 monoclonal antibodies (MoAb) CR11-351 and 4B inhibit the binding of each other to HLA-A2 lymphoid cells and block the cytotoxicity of the anti-HLA-A2 cytotoxic-T-cell clone R32. The blocking does not reflect reactivity of the MoAb CR11-351 and 4B and of the cytotoxic-T-cell clone R32 with the same determinant, since they display differential reactivity with four HLA-A2 variants which carry amino acid substitutions at different positions. These results show for the first time in the human system that Class I HLA variants represent useful reagents to compare the fine specificities of monoclonal antibodies and T-cell clones. Furthermore our data suggest that T-cell recognition depends upon the tertiary structure of the antigen.

Antibodies, Monoclonal↗
PubMed1984

Monoclonal antibodies directed against human Ia antigens detect an evolutionary conserved epitope on guinea pig Ia antigens with unique functional properties.

Three murine monoclonal antihuman Ia antibodies were found to cross react with guinea pig Ia antigens. Sequential immunoprecipitation studies demonstrated that these antibodies recognized a subset of guinea pig Ia antigens. In competition binding studies, the antihuman Ia antibodies competed with each other, but not with a number of murine monoclonals raised directly to guinea pig Ia antigens. When the antihuman Ia monoclonals were tested for their ability to inhibit the proliferative response of primed guinea pig T cells to antigen-pulsed macrophages, they completely inhibited the response of strain 2 T cells to the complex antigen ovalbumin while having no effect on the response to the copolymer L-glutamic acid, L-lysine. In contrast, the monoclonals raised against guinea pig Ia had much less dramatic effects on the response of uncloned T cells to these antigens. These results suggest that the cross-reactive monoclonals may recognize an evolutionary conserved epitope on Ia that has a regulatory function in T-cell activation.

Animals↗
PubMed1984

HLA class I specific T lymphocyte clones with dual alloreactive functions.

Four human T lymphocyte clones exhibiting proliferative responses to class I HLA antigens were isolated from an in vitro mixed lymphocyte culture (MLC). Three clones expressed the Leu-2+3- phenotype and demonstrated proliferation in response to HLA-B8, while the fourth clone expressed the Leu-2-3+ phenotype and proliferated in response to HLA-A2. These clones were also cytotoxic towards cells bearing the same target antigens. Blocking studies utilizing monoclonal antibodies demonstrated that proliferation was triggered by determinants on the class I molecule itself, and these determinants appear to be spatially close to those which determine serologic allospecificity. These findings support the concept that the class I molecules themselves are the weak MLC stimulating determinants previously mapped to the HLA-A and B regions of the major histocompatibility complex.

Cells, Cultured↗