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S Ferrone

Publications and source records attributed to S Ferrone.

At least 217 records · Page 12Linked to original sources
PubMed1988

A solid-phase immunoassay to screen anti-HLA monoclonal antibodies.

A solid-phase assay to detect anti-HLA monoclonal antibodies was developed. In this assay microtiter plates are coated with antigens solubilized from cultured lymphoid cells by sonication and then incubated with anti-HLA monoclonal antibodies. The antigen-antibody interaction is indicated by the development of color following the addition of peroxidase-conjugated anti-mouse Ig xenoantibodies and its substrate. The assay is rapid since it does not require centrifugations during the washing steps. Furthermore the assay is simple, reproducible and suitable to screen large numbers of samples and to detect antibodies recognizing determinants not exposed on the membrane of viable cells. The sensitivity of the assay is influenced by the pH of the buffer used to coat plates with antigens, by the number of cells used to prepare soluble antigens, by the incubation time of antigen preparations with plates and by the incubation time of antibody preparations with antigen-coated plates. Titration of anti-HLA monoclonal antibodies with known specificity and screening of hybridomas generated with splenocytes from mice immunized with cultured human lymphoid cells indicate that the sensitivity of the solid-phase assay is similar to that of the ELISA with lymphoid cells.

Antibodies, Monoclonal↗
PubMed1988

Level of HLA antigens in locoregional metastases and clinical course of the disease in patients with melanoma.

Immunohistochemical staining with monoclonal antibodies showed marked variations in the percentage of melanoma cells stained by anti-HLA Class I and anti-HLA Class II monoclonal antibodies among 48 locoregional metastases removed from 39 patients with malignant melanoma. On the other hand there was limited variation in the percentage of melanoma cells stained by anti-HLA antibodies in autologous locoregional metastases removed from 8 of 9 patients. In the remaining patient marked differences were found in the percentage of melanoma cells stained by anti-HLA Class I antibodies in the two parts of the lymph node metastasis analyzed. Therefore this patient was not included in additional analyses to correlate the level of expression of HLA antigens with the clinical course of the disease. In all the lesions tested the percentage of melanoma cells stained by anti-HLA Class II antibodies was lower than or equal to but never higher than that stained by anti-HLA Class I antibodies. According to the level of expression of HLA Class I and Class II antigens the 38 patients could be divided into three groups: Pattern A included lesions with more than 50% of tumor cells stained by anti-HLA Class I antibodies (mean, 86.1; median, 85) and 50% or less by anti-HLA Class II antibodies (mean, 10.5; median, 5); Pattern B included lesions with 50% or less tumor cells stained by anti-HLA Class I antibodies (mean, 14.9; median, 5) and by anti-HLA Class II antibodies (mean, 4.1; median, 1); Pattern C included lesions with more than 50% tumor cells stained by anti-HLA Class I antibodies (mean, 88.8; median, 92) and by anti-HLA Class II antibodies (mean, 70.0; median, 70). The survival of 21 patients with Pattern A was significantly longer than those of 13 and 4 patients with Patterns B and C, respectively. No difference in the survival of patients in the latter two groups was found. These results suggest that HLA antigens play a role in the biology of melanoma and that analysis of the level of HLA antigens in locoregional metastases of patients with melanoma may provide clinically useful information.

Adult↗
PubMed1988

Comparison of features of carcinogen-transformed human cells in vitro with sarcoma-derived cells.

To define characteristics of chemically transformed phenotypes during and after progression to neoplasia and to assess their relationship to those phenotypes expressed by surgically removed sarcoma lesions, we compared the characteristics in the following manner. We investigated: (1) alterations in growth patterns; (2) anchorage-independent growth; (3) reactivity with monoclonal antibodies directed against surface antigen; (4) invasiveness in embryonic chick skin; (5) tumorigenicity in nude mice; and (6) karyology. Fifty different sarcoma cell lines were examined which exhibited different rates and absolute numbers of population doublings. With one exception, all sarcoma cell lines exhibited a finite life span ranging from 60 to 100 population doublings. Populations of these cells that exhibited anchorage-independent growth in soft agar also reacted positively with a monoclonal antibody (MoAb) 345.134S directed against a 115K-GP cell surface glycoprotein. Similarly, chemically transformed cells that grew in soft agar also reacted with the MoAb 345.134S, whereas cells with an inability to grow in soft agar did not. Cell lines established from human sarcoma and from chemically transformed human fibroblasts that reacted positively with the MoAb 345.134S were invasive for embryonic chick skin and formed tumors in nude mice. The selection medium used during culture of the carcinogen-treated cells resulted in the appearance of an altered phenotype that after at least 16 population doublings exhibited characteristics common to those cells derived from human sarcomas.

Animals↗
PubMed1988

Syngeneic antiidiotypic monoclonal antibodies to the murine anti-HLA-DR,DP monoclonal antibody CR11-462.

Two out of 625 hybridomas constructed with splenocytes from a Balb/c mouse immunized with the murine anti-HLA-DR, DP monoclonal antibody CR11-462 secrete antiidiotypic antibodies. Binding assays with a panel of 12 anti-HLA class I, 14 anti-HLA class II, and 9 anti-human melanoma-associated antigen MoAbs showed that the idiotopes recognized by MoAb F3-C25 and F3-B6 are not expressed by any of the tested antibodies, even those which inhibit the binding of MoAb CR11-462 to cultured B lymphoid cells. Crossblocking experiments showed that the two idiotopes are distinct and spatially distant. Both idiotopes require the association of heavy and light chain of MoAb CR11-462 for their expression. The idiotope recognized by MoAb F3-B6 is an alpha idiotope, since it is located outside the antigen combining site of MoAb CR11-462. The idiotope recognized by MoAb F3-C25 is a gamma idiotope, since it is antigen inhibitable but does not possess an internal image of the antigen. Interestingly the MoAb F3-C25 displays a differential inhibitory effect on the binding of MoAb CR11-462 to lymphoid cells that express different HLA phenotypes or share the HLA-DRw6 allospecificity. These results suggest that antiidiotypic antibodies may sharpen our ability to dissect the heterogeneity of HLA class II antigens.

Animals↗
PubMed1988

Anti-HLA-B7,B27,Bw42,Bw54,Bw55,Bw56, Bw67,Bw73 monoclonal antibodies: specificity, idiotypes, and application for a double determinant immunoassay.

The monoclonal antibodies (MoAbs) KS3 and KS4 are secreted by hybridomas constructed with splenocytes from a BALB/c mouse sequentially immunized with the cultured lymphoid cells JKu and LG-2 which share only the HLA-B27 specificity. Serologic and immunochemical assays have shown that the two MoAbs recognize the same (or spatially close) determinant expressed by HLA-B7,B27,Bw42,Bw54,Bw55,Bw56,Bw67, and Bw73 alloantigens. This determinant is spatially close but distinct from those defined by the anti HLA-B27 monoclonal antibodies described in the literature. The syngeneic antiidiotypic MoAb T12-105 and T12-211 elicited with MoAb KS4 were shown to recognize idiotopes within the antigen combining site of MoAb KS3 and KS4. Neither idiotope was detected on the anti HLA class I and anti HLA class II monoclonal antibodies tested. The MoAb KS4 in combination with the anti human beta 2-microglobulin MoAb NAMB-1 was utilized to develop a double determinant immunoassay (DDIA). The latter represents a sensitive method to detect and quantitate HLA-B27 antigens in spent culture medium of lymphoid cell lines and in serum. Typing for HLA-B27 antigens with the DDIA of sera from HLA typed donors yielded results highly correlated with those of the conventional lymphocytotoxicity assay.

Antibodies, Monoclonal↗
PubMed1988

Fine specificity and idiotype diversity of the murine anti-HLA-A2, A28 monoclonal antibodies CR11-351 and KS1.

The monoclonal antibodies (MoAb) CR11-351 and KS1 are secreted by hybridomas generated with splenocytes from BALB/c mice immunized with the cultured human B lymphoid cells LG-2 (HLA-A2,A2,B27,B27). The 2 monoclonal antibodies immunoprecipitated components with a superimposable 2-dimensional gel electrophoretic profile from the cultured B lymphoid cells LG-2 and inhibited the cytotoxicity of the anti-HLA-A2,A28 T cell clone MI#3 to a similar extent. In crossinhibition experiments, the MoAb CR11-351 and KS1 completely inhibited the binding of each other to lymphoid cells with the appropriate HLA phenotype. Testing with a panel of HLA-typed lymphoid cells showed that the MoAb CR11-351 and KS1 display the same serologic specificity, since both of them react with HLA-A2 and/or A28 antigens bearing cells. The 2 monoclonal antibodies recognize distinct, although spatially close, determinants, since only the MoAb CR11-351 displays differential reactivity with HLA-A2 variant cell lines and reacts with HLA-A9 bearing B lymphoid cells. Analysis of MoAb CR11-351 and KS1 with syngeneic polyclonal and monoclonal antiidiotypic antibodies detected no sharing of idiotopes between the 2 monoclonal antibodies. In view of their reactivity with distinct determinants, these results are in agreement with the concept that the antigenic specificity of an antibody controls at least in part the expression of its idiotypes.

Animals↗
PubMed1988

Differential expression of HLA-DR, DQ, and DP antigens in primary and metastatic melanoma.

Fifty-five primary and 33 metastatic surgically removed melanoma lesions were stained in indirect immunoperoxidase with anti HLA-DR, DQ, and DP monoclonal antibodies and with the monoclonal antibody CL203.4 to a 96-K melanoma associated antigen (MAA). The latter antigen may represent a marker to monitor susceptibility of melanoma cells to modulation by IFN-gamma, because it is highly susceptible to induction by IFN-gamma. In primary melanomas 44%, 29%, 10%, and 55% of the lesions tested were evidently stained by anti HLA-DR, DQ, DP, and 96-K MAA monoclonal antibodies, respectively. A statistically significant association (P less than 0.01) was demonstrated between the degree of intratumoral lymphocytic infiltrate and the expression of HLA-DR and HLA-DQ antigens. In addition, a high degree of concordance in the reactivity pattern of individual lesions stained for HLA-DR antigens and for the 96-K MAA was found. In metastases 64%, 33%, 47%, and 100% of the lesions tested were evidently stained by anti HLA-DR, DQ, DP, and 96-K MAA monoclonal antibodies, respectively. This study indicates that HLA-DR and HLA-DP antigens are expressed in a higher percentage of metastatic than of primary melanomas and that there is no marked difference in the expression of HLA-DQ antigens between primary and metastatic melanomas. The data suggest that the regulatory mechanisms which control the expression of HLA-DR and DP antigens in primary and metastatic melanoma lesions are different. Locally produced IFN-gamma may play a role in the regulation of HLA Class II antigens in primary melanomas.

Antibodies, Monoclonal↗
PubMed1987

Differential regulatory role of monomorphic and polymorphic determinants of histocompatibility leukocyte antigen class I antigens in monoclonal antibody OKT3-induced T cell proliferation.

Monoclonal antibodies (mAb) to monomorphic and polymorphic determinants on the heavy chain of histocompatibility leukocyte antigen (HLA) class I antigens inhibit mAb OKT3-induced T cell proliferation, whereas the anti-beta 2-microglobulin mAb NAMB-1 does not affect it. The inhibitory effect of anti-HLA class I mAb is specific, is not an Fc-mediated phenomenon, does not require accessory cells, and does not involve early stages of T cell activation. Distinct determinants of HLA class I antigens regulate T cell proliferation by different mechanisms, because the anti-HLA-A2, A28 mAb CR11-351, and the mAb W6/32 to a framework determinant of HLA class I antigens block interleukin 2 (IL-2) secretion and IL-2 receptor expression, whereas the mAb CR10-215 to a monomorphic determinant blocks only IL-2 receptor expression. The mAb CR10-215 and W6/32 induced a 50% of maximal inhibition of T cell proliferation, when added after 27 and 12 hr, respectively, of incubation of peripheral blood mononuclear cells with mAb OKT3. On the other hand, the mAb CR11-351 inhibited T cell proliferation even when added after 38 hr of incubation of peripheral blood mononuclear cells with mAb OKT3 and was the only one to inhibit proliferation of cycling T lymphocytes. It is suggested that HLA class I antigens regulate T cell proliferation by interacting with cell-surface molecules involved in T cell activation. The differential inhibitory activity of the anti-HLA class I monoclonal antibodies tested may reflect the different ability of the corresponding determinants to interact with activation molecules.

Antibodies, Monoclonal↗
PubMed1987

Immunogenicity of human melanoma-associated antigens defined by murine monoclonal antibodies in allogeneic and xenogeneic hosts.

The immunogenicity in patients with melanoma, in monkeys, and in rabbits of four human melanoma-associated antigens (MAA) defined by murine monoclonal antibodies was investigated. The latter included the high molecular weight MAA and the (Mr 115,000, 100,000, and 95,000-150,000 MAA. To this end sera from patients with melanoma, from monkeys, and from rabbits immunized with cultured human melanoma cells were tested for their ability to inhibit the binding to cultured human melanoma cells of radiolabeled anti-Mr 95,000-150,000 MAA monoclonal antibody (MoAb) 140.72, anti-high molecular weight MAA MoAb 225.28, anti-Mr 115,000 MAA MoAb 345.134, and anti-Mr 100,000 MAA MoAb 376.96. None of the sera from patients with melanoma significantly inhibited the reactivity of any of the anti-MAA monoclonal antibodies with melanoma cells. Of the sera from the six monkeys immunized with human melanoma cells, two sera significantly inhibited the reactivity with cultured human melanoma cells of both MoAb 345.134 and 376.96, one serum inhibited only that of MoAb 345.134, and the remaining three sera did not inhibit any of the four anti-human MAA monoclonal antibodies. Sera from six of the seven rabbits immunized with cultured human melanoma cells inhibited the binding to melanoma cells of at least one of the four anti-human MAA monoclonal antibodies while the serum from one rabbit immunized with a melanoma cell extract had no effect. Marked differences were found among the individual rabbit sera in their ability to inhibit the binding of the four anti-human MAA monoclonal antibodies. Sequential immunoprecipitation experiments corroborated the serological findings obtained with one of the two rabbit antisera tested. These results suggest that the immunogenicity of human MAA in mice may be different from that in patients with melanoma and in other animal species.

Animals↗
PubMed1987

Immunosuppressive activity of T cell clones generated from human T cells stimulated with autologous TPHA cells.

T cell clones were generated from human T cells stimulated with autologous phytohemagglutinin (PHA)-activated T (TPHA) cells. Characterization of three T cell clones originated from donor SF and one from donor JM showed that they proliferated when stimulated with autologous TPHA cells, non-T cells, and peripheral blood mononuclear cells, but did not proliferate when stimulated with allogeneic TPHA cells, non-T cells, and mononuclear cells, with autologous and allogeneic resting T cells, and with PHA. These results in conjunction with the blocking of the proliferation by anti-histocompatibility leukocyte antigen class II monoclonal antibodies indicate that these class II antigens are involved in the proliferation of T cell clones stimulated with autologous lymphoid cells. The four T cell clones are cytotoxic neither to autologous lymphoid cells nor to a panel of cultured human cell lines. The four T cell clones display immunosuppressive activity, since they inhibit the proliferation of autologous and allogeneic cells stimulated with antigens and mitogens and the secretion of immunoglobulin by B cells stimulated with pokeweed mitogen in presence of T cells. Furthermore, the four T cell clones display differential inhibitory activity on the proliferation of cultured human cell lines. The immunosuppressive activity is species-specific, since the T cell clones do not inhibit the proliferation of murine cells. The suppression is mediated by a factor(s) with an apparent m.w. of 13,000 to 16,000. The suppressor activity is labile at alkaline pH and is lost following incubation with pronase (100 U/ml) for 30 min at 37 degrees C.

Antibodies, Monoclonal↗
PubMed1987

A cytoplasmic human melanoma associated antigen as a marker of activation in lymphoid cells.

A cytoplasmic glycoprotein, originally identified by the monoclonal antibody 465.12S in melanoma tumors, is significantly increased in epithelial cells of different histotype following transformation. In the present study we show that the cytoplasmic melanoma associated antigen (cyt-MAA) is drastically enhanced in lymphoid cells by polyclonal and allogeneic stimulation, as well as by transformation. Normal T-cells with helper and suppressor phenotype are far more susceptible than B-cells to this enhancement. However, among transformed lymphoid cells, the expression of the cyt-MAA does not correlate with lineage, but rather with stage of differentiation. Acute lymphoblastic leukemias represent the only exception, since in these lymphoid malignancies cyt-MAA levels are highly heterogeneous even within groups of phenotypically similar lesions. Thus, the expression of the cyt-MAA is shared by cells of distant embryological origin in early stages of their differentiation and/or during proliferation. Quantitation of the cyt-MAA may provide useful information for the classification of some lymphoid malignancies.

Antigens, Neoplasm↗
PubMed1987

Characterization of a monoclonal antibody-defined human melanoma-associated antigen susceptible to induction by immune interferon.

To develop monoclonal antibodies (mAb) recognizing human melanoma-associated antigens (MAA) susceptible to modulation by immune interferon (IFN-gamma), hybridomas were constructed with splenocytes from a BALB/c mouse immunized with IFN-gamma-treated melanoma cells Colo 38. Screening of supernatants with control and IFN-gamma-treated melanoma cells showed that the mAb CL203 and CL207 display preferential reactivity with IFN-gamma-treated melanoma cells. The two mAb recognize the same (or spatially close) determinant on a 96,000 MAA which has a density of 0.36 X 10(6) antigenic sites/cell on untreated melanoma cells Colo 38 and of 1.39 X 10(6) and 1.54 X 10(6) on melanoma cells Colo 38 treated with IFN-gamma (final concentration, 200 U/ml) for 24 and 48 hr, respectively. The effect of IFN-gamma on the 96,000 MAA is dose- and time-dependent, reversible, and blocked by inhibitors of RNA and protein synthesis. Furthermore, the effect of IFN-gamma on the induction of the 96,000 MAA appears to be specific, inasmuch as IFN-alpha and IFN-beta do not induce the expression of the 96,000 MAA. The latter is also induced by IFN-gamma in a variety of carcinoma cell lines, but its level is markedly lower than on melanoma cells. Furthermore, the apparent m.w. of the antigen synthesized by the carcinoma cell lines in the presence of IFN-gamma ranges between 93,000 and 96,000. This molecular heterogeneity appears to reflect differences in the degree of glycosylation of the polypeptide moiety because the antigen synthesized by a variety of cell lines in the presence of tunicamycin has an apparent m.w. of 51,000.

Antibodies, Monoclonal↗
PubMed1987

Cross-reactivity of murine anti-human high molecular weight-melanoma associated antigen monoclonal antibodies with guinea pig melanoma cells.

To identify melanoma associated antigens (MAAs) shared by human and guinea pig melanoma cells, a battery of murine monoclonal antibodies (MoAbs) to human MAA and an antiserum to S100 protein were tested with four newly established guinea pig melanoma cell lines. Only the monoclonal antibodies 149.53 and 225.28 which recognize distinct determinants of the human high molecular weight-MAA (HMW-MAA) reacted with all four guinea pig melanoma cell lines. To compare the binding site of MoAbs 149.53 and 225.28 with guinea pig and human melanoma cells, inhibition binding experiments were performed with antiidiotypic monoclonal antibodies which completely inhibit the binding of MoAbs 149.53 and 225.28 to human melanoma cells. The binding of MoAb 149.53 to guinea pig melanoma cells was partially inhibited by antiidiotypic MoAbs MF9-10 and MK1-180 which recognize distinct private idiotopes within the antigen combining site of MoAb 149.53. On the other hand the binding of MoAb 225.28 to guinea pig melanoma cells was completely inhibited by antiidiotypic MoAbs MF11-30 and TK1-F2 which recognize distinct private idiotopes within the antigen combining site of MoAb 225.28. These results suggest that the determinant recognized by MoAb 149.53 on guinea pig melanoma cells is similar but not identical to that recognized on human melanoma cells, while the determinants recognized by MoAb 225.28 on the two types of cells do not display any detectable differences under the experimental conditions tested. The target structure on the guinea pig melanoma cells identified by MoAbs 149.53 and 225.28 is a Mr 280,000 molecule which has the same apparent molecular weight as one of the two subunits of the HMW-MAA synthesized by human melanoma cells. Sequential immunoprecipitation experiments with guinea pig melanoma cells showed that the determinant recognized by MoAb 149.53 is expressed on a subpopulation of the molecules recognized by MoAb 225.28. Immunohistochemical staining with MoAb 225.28 of a variety of different tissues from normal adult guinea pigs showed that the corresponding antigenic determinant is detectable only in basal cells of epidermis and hair follicles of skin. S100 protein, which is a cytoplasmic constituent of normal human melanocytes, benign nevi, and malignant melanocytes, was also detected in the cytoplasm of the four cultured guinea pig melanoma cells lines. The results of the present investigation may lead to a better understanding of the phylogenetic evolution of the human HMW-MAA and suggest that guinea pig melanoma may serve as a useful animal model for immunobiological studies and carcinogen-induced tumorigenesis investigations.

Animals↗
PubMed1987

Syngeneic antiidiotypic antisera to murine antihuman high-molecular-weight melanoma-associated antigen monoclonal antibodies.

BALB/c mice were immunized with the murine antihuman high-molecular-weight melanoma-associated antigen monoclonal antibodies (MoAbs) 149.53, 225.28, 653.25, and 763.74. Antiidiotypic antibodies could be detected in bleedings obtained 3 days following the first booster, increased in titer in bleedings obtained following the second booster, and persisted at high level in bleedings obtained 38 days following the second booster. Cross-blocking studies with a panel of anti-melanoma-associated antigen, anti-HLA Class I, and anti-HLA Class II monoclonal antibodies showed that the antisera recognize private idiotypes. The latter are located within the antigen combining site, since antiidiotypic antisera specifically inhibited the binding of the corresponding immunizing anti-human high-molecular-weight melanoma-associated antigen monoclonal antibody to cultured human melanoma cells Colo 38 in a dose-dependent fashion. The spectrotype of the anti-MoAb 149.53 antiserum comprises eight major components in the range of pH 6.2 to 7.0; those of the anti-MoAb 225.28 antiserum and of the anti-MoAb 653.25 antiserum, two major components in the ranges of pH 6.4 to 6.6 and 6.5 to 6.7, respectively; and that of the anti-MoAb 763.74 antiserum, three major components in the range of pH 6.2 to 6.4.

Animals↗
PubMed1987

Syngeneic anti-idiotypic antisera to murine anti-HLA class II monoclonal antibodies.

BALB/c mice have been immunized with six anti-HLA Class II monoclonal antibodies (MoAb); the latter included MoAb CR11-462, Q5/6, and Q5/13 to monomorphic determinants, the anti-HLA-DR1,4,w6,w8,w9 MoAb AC1.59, the anti-HLA-DRw9 MoAb AB7ae9, and the anti-HLA-DQw3 MoAb AC6G. The six monoclonal antibodies markedly differ in their ability to induce anti-idiotypic antibodies, because the latter were not detected in the sera from the mice immunized with the MoAb AB7ae9 and with the MoAb AC6G. The MoAb AC1.59 and CR11-462 elicited antibodies to private idiotypes, and the MoAb Q5/6 and Q5/13 elicited antibodies to private and public idiotypes. The titer of the latter in the anti-MoAb Q5/6 antiserum appears to be markedly lower than that of the former ones; no marked difference was detected in the titer of the two types of antibodies in the anti-MoAb Q5/13 antiserum. Blocking experiments with a panel of monoclonal antibodies showed that the MoAb Q5/13 shares idiotypes with the anti-HLA Class I MoAb Q5/6, 127, and 441 and with the anti-HLA Class I MoAb CR11-351, Q1/28, Q6/64, and 6/31, but does not share idiotypes with any of the nine anti-human melanoma-associated antigen MoAb tested. The spectrotypes of the anti-MoAb CR11-462 and anti-MoAb Q5/13 antisera comprise two major components in the pH 6.2 to 6.7 range, that of the anti-MoAb Q5/6 antiserum comprises two major components in the pH 6.5 to 6.8 range, and that of the anti-MoAb AC1.59 antiserum comprises a number of components in the pH 5.6 to 7.2 range.

Animals↗