Structural polymorphism of human Ia antigens recognized by the monoclonal antibody I2 and by cross-reacting I-Ek-specific alloantibodies.
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Amino Acid Sequence↗
Biomedical subjects
S Ferrone
Publications and source records attributed to S Ferrone.
Differential tissue distribution and ontogeny of DC-1 and HLA-DR antigens.
The tissue distribution and the ontogeny of DC-1 antigens have been investigated and compared with those of HLA-DR antigens. Indirect immunofluorescence (IIF) staining of surgically removed normal tissues from adults with the monoclonal antibody (MoAb) BT3.4 has detected DC-1 antigens in tissues of various embryologic origin. The tissue distribution of DC-1 antigens is more restricted than that of HLA-DR antigens, as the former are not detected in duodenal epithelium, colon mucosa, and ductal mammary gland epithelium. In fetuses up to 26 weeks of age, DC-1 antigens were detected only on cortical and medullary thymic dendritic cells with an anatomic distribution similar to that of reticuloepithelial cells and in endothelial cells of the small intestine. At this stage of intrauterine life, HLA-DR antigens have already reached their full tissue distribution. The tissue distribution and the ontogeny of DC-1 antigens resemble those of their murine counterparts, i.e., the I-A antigens.
Adult↗
Ia antigen expression is increased on tunicamycin-resistant human B-lymphoid cells.
Cultured human B-lymphoid cells WIL-2 were mutated with ethylmethane sulfonate and selected for resistance to tunicamycin, an antibiotic that selectively inhibits N-linked glycosylation. Ultrastructural analysis of five isolated tunicamycin-resistant mutants (TMR) showed changes in surface microvilli, dilation of the endoplasmic reticulum, and an increase in surface myelin figures. Cytofluorometric analysis of TMR cells incubated with anti-HLA monoclonal antibodies showed a normal density of HLA-A, B antigens and an increased density of Ia antigens. The properties of these TMR mutants have remained stable for at least 18 months. These TMR cells may serve as a useful model to study the biochemical events in the processing and expression of Ia antigens.
Antibodies, Monoclonal↗
Analysis of the repertoire of anti-HLA antibodies with anti-idiotypes to a murine anti-HLA-A2,A28 monoclonal antibody.
Xenoantibodies to idiotypes of the anti-HLA-A2,A28 MoAb CR11 -351 were isolated from an antiserum raised in rabbit #81 by immunizations with purified MoAb CR11 -351. The purification procedure involved absorption with insolubilized mouse immunoglobulins and monoclonal antibodies and affinity chromatography on insolubilized MoAb CR11 -351. Two antibody populations were identified in the xenoantibody preparation #81: one recognizes a recurrent idiotope expressed by the MoAb CR10 -215, CR10 -402, Q1/28, Q6 /64, and 6/31 to monomorphic determinants of HLA-A,B antigens and by the MoAb CR10 -343, CR11 -462, and Q5/6 to human Ia antigens. The other antibody population recognizes a private idiotope. Neither idiotope was detected on the anti-HLA-A2,A28 variant (A28) MoAbs BB7 .2, MA2 .2, and PA2.1, on the anti-HLA-A2,B17 MoAb MA2 .1 and on antibody populations in conventional anti-HLA-A2,A28 antisera. The idiotopes were also not detected on the anti-HLA-A2,A28 MoAb A2,A28 M1 which recognizes a determinant spatially close to that identified by the MoAb CR11 -351. The idiotope(s) recognized by the xenoantibodies #81 may be located in the combining site of the MoAb CR11 -351, since its incubation with the anti-idiotype antibodies specifically blocks the reactivity with lymphoid cells with the appropriate HLA phenotype.
Animals↗
Heterogeneity of human peripheral blood mononuclear cells detected by monoclonal antibodies to monomorphic determinants of human Ia antigens.
The reactivity patterns of several monoclonal antibodies specific for monomorphic determinants of human Ia antigens were studied using flow cytometric techniques. We observed differential reactivity of these antibodies with human lymphoid cell lines, normal fresh human mononuclear cells, and lymphoblasts from PHA-activated cultures. The molecular heterogeneity of Ia antigens previously identified with immunochemical techniques was accompanied by heterogeneity of cell surface expression as identified by an immunofluorescent probe. The determinants identified by these anti-Ia monoclonal antibodies may provide useful markers in the isolation of cellular subpopulations responsive in the immune system.
Antibodies, Monoclonal↗
The early posttransplant prognosis of acute renal allograft rejection as determined by detection of cytotoxic antibodies to lymphoid B cell lines.
We have studied serial samples of pretransplant and posttransplant sera for cytotoxic antibodies to lymphoid B cell lines (LCL) in 45 renal allograft recipients. A total of 48 rejection reactions occurred in 31 patients. A comparison of each patient's most reactive posttransplant serum showed a significantly higher reactivity in the ten patients with early allograft failure when compared with the 21 patients with reversible rejections and the 14 patients who had no rejections. Rejection reactions were easily differentiated by comparing the change in cytotoxic reactivity to LCL of recipients' sera drawn at the time of a rejection episode with the reactivity of their pretransplant sera. In 32 rejections considered non-antibody-associated cytotoxic reactivity of recipients' sera to LCL either decreased or remained essentially unchanged during the rejection. In 16 rejections considered antibody-associated the recipients' sera drawn during the rejection episode showed an increase in cytotoxic reactivity ranging from 40% to 100%. Response to antirejection therapy and three month graft survival had a significant correlation with changes in LCL antibody reactivity during a rejection. Only two of the 32 rejections considered non-antibody-associated failed to reverse compared with eight of the 16 antibody-associated rejections (P less than .001). Graft survival at three months in patients with non-antibody-associated rejections was 90% compared with 27% in the 11 patients who had antibody-associated rejections (P less than .001) Other parameters possibly related to the severity of a rejection reaction or to early allograft prognosis did not differ appreciably between the two types of rejections. This included the time posttransplant to the first rejection episode, the number of patients with multiple rejections in the first three months, and rejections requiring dialysis therapy. Determination of a change in cytotoxic reactivity to LCL during a rejection reaction enables one to predict the response to antirejection therapy and early allograft prognosis. This may ultimately be useful in selecting different types of antirejection therapy for individual patients.
Acute Disease↗
Cross reaction of monoclonal antibodies to human MHC class I and class II products with bovine lymphocyte subpopulations.
Peripheral blood lymphocytes (PBL) from cattle have been separated into T & B cell subpopulations using a panning technique. These T and B cell preparations have then been tested by direct and indirect complement mediated cytotoxicity tests with a series of monoclonal antibodies (Moabs) reacting with HLA class I and class II products. The anti-class I monoclonals were monomorphic or non-reactive and where reactive killed both B and T cells. The anti-class II monoclonals reacted only with B lymphocytes. There was no relationship between their reactivity pattern and any BoLA specificity.
Animals↗
Immunohistochemical analysis of malignant melanomas and nevocellular nevi with monoclonal antibodies to distinct monomorphic determinants of HLA antigens.
Using an indirect immunoperoxidase technique, 20 nevocellular nevi, 5 dysplastic nevi, 14 primary cutaneous melanomas, and 24 metastatic melanomas were tested with a panel of monoclonal antibodies to monomorphic determinants of Class I (HLA-A,B,C) and Class II (la-like) major histocompatibility complex antigens. Class I HLA and beta 2-microglobulins were not detected on the majority of nevus cells but were expressed by 3 of 5 dysplastic nevi, by the majority of tumor cells in 12 of 14 primary cutaneous melanomas, and in 13 of 24 metastases. The different expression of Class I HLA and beta 2-microglobulins in primary and metastatic lesions suggests that loss of these antigens may be associated with progression of malignancy. Class II HLA were not detected in common nevi but were locally present in 1 of 5 dysplastic nevi, 7 of 14 cases of primary cutaneous melanoma, and all 24 cases of metastatic lesions tested. These findings suggest that increase in Class II HLA expression may be associated with progression of malignancy. The staining patterns obtained with monoclonal antibodies to distinct determinants of Class I HLA and Class II HLA were superimposable within each type of antigen. Therefore, the discrepancies in the literature about the expression of histocompatibility antigens by lesions of melanocytic origin are not likely to reflect the different specificity of the antibodies used by the various investigators.
Adolescent↗
Phenotyping of lesions of melanocyte origin with monoclonal antibodies to melanoma-associated antigens and to HLA antigens.
The antigenic profiles of a large number of surgically removed human benign and malignant lesions of melanocyte origin have been analyzed with the use of monoclonal antibodies (MoAb) against la antigens, against the HLA-A,B,C-beta 2-microglobulin molecular complex, against a cytoplasmic melanoma-associated antigen (MAA), and against membrane-bound MAA. Membrane-bound MAA include a high-molecular-weight MAA (HMW-MAA), a 115K MAA, and a 100K MAA. Appearance of the HMW-MAA and of the cytoplasmic MAA, as well as cytoplasmic distribution or loss of HLA-A,B,C antigens, occurs in benign lesions. Additional appearance of Ia antigens is associated with malignant transformation of melanocytes. The antigenic profile defined by the battery of MoAb used displays differences among benign lesions of different histogenesis, between benign and malignant lesions, and among malignant lesions with different histopathologic properties. These results suggest that phenotyping of surgically removed lesions with anti-MAA and anti-HLA MoAb may contribute to the understanding of the steps involved in tumor progression of melanocytes and may aid in the diagnosis of lesions with unusual histopathologic features.
Antibodies, Monoclonal↗
Early determination of renal allograft survival: a correlation between cytotoxic antibodies in recipient sera, renal pathology, and response to antirejection therapy.
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Clinical Trials as Topic↗
A comparison of the cyclic anhydride and mixed anhydride methods for 111In-DTPA chelation to monoclonal antibodies.
The cyclic anhydride (CA) and the mixed anhydride (MA) of DTPA were synthesized and used to chelate 111In to an antimelanoma monoclonal antibody. The CA and MA methods showed mean labeling efficiencies of 25.7 and 20.5%, respectively (p = NS). The binding efficiency of labeled antibody to human melanoma cells in tissue culture also was similar (means = 52 and 50%, respectively, p = NS), as was tumor uptake in nude mice at 96 hrs post-injection (16%-CA vs 12%-MA). The method required less complicated chemical syntheses, much less preparation time, and the product was stable over a much longer period. The results suggest that the CA method is preferable for bifunctional chelate labeling of monoclonal antibodies with 111In-DTPA.
Animals↗
Complement-dependent killing of human hematopoietic progenitor cells with noncomplement-fixing monoclonal antibodies in an antiglobulin assay.
A complement (C)-dependent antiglobulin assay was utilized to determine the reactivity of non-C-fixing monoclonal antibodies (MoAb) with human granulocyte-macrophage progenitor cells (CFU-GM). The variables of the assay were analyzed with non-C-fixing MoAb against Ia antigens, including CR11-462, which recognizes the same (or spatially close) determinant identified by the C-fixing anti-Ia MoAb Q5/13. The sensitivity of the antiglobulin assay was influenced by dilutions of anti-mouse Ig xenoantiserum and of rabbit C. Five non-C-fixing MoAb to Ia antigens, seven non-C-fixing MoAb to HLA-A,B antigens, and one non-C-fixing MoAb to beta 2-microglobulin induced marked inhibition of human CFU-GM in the antiglobulin assay. The activity of non-C-fixing MoAb in the antiglobulin assay was comparable to that of C-fixing anti-Ia and anti-HLA-A,B MoAb in the standard cytotoxicity assay. In addition, the cytotoxic effect of dilute C-fixing anti-Ia MoAb was enhanced when the antiglobulin technique was employed. The results of this study indicate that the antiglobulin assay is a rapid and simple technique for the characterization of antigens on human hematopoietic progenitors. Our data also indicate that Ia antigens are expressed on most CFU-GM and that the conflicting results in the literature (that is, those suggesting that Ia antigens are expressed on a smaller proportion of CFU-GM) may reflect differences in the cytolytic activity of the MoAb and rabbit C used.
Adult↗
Distribution of human Class I (HLA-A,B,C) histocompatibility antigens in normal and malignant tissues of nonlymphoid origin.
Indirect immunofluorescence and immunoperoxidase staining of surgically removed tissues of nonlymphoid origin with monoclonal antibodies to the heavy and light chain of HLA-A,B,C antigens have shown that they have a more restricted tissue distribution than previously assumed. HLA-A,B,C antigens were not detected in brain cortex, cerebellum, sympathetic ganglia, hypophysis, parathyroid gland, thyroid, exocrine pancreas, hepatocytes, sperm, seminiferous tubules, or skeletal or smooth muscle. Malignant transformation of cells may be associated with appearance, changes in cellular distribution of HLA-A,B,C antigens, and/or dissociation in the expression of the two subunits. Analysis of primary tumors and of autologous metastases showed heterogeneity in the expression of HLA-A,B,C antigens among lesions removed from different sites. The degree of heterogeneity did not correlate with the site of origin of metastases.
Female↗
Level of a membrane-bound high-molecular-weight melanoma-associated antigen and a cytoplasmic melanoma-associated antigen in surgically removed tissues and in sera from patients with melanoma.
Utilizing double-determinant immunoassays (DDIAs), the high-molecular-weight melanoma-associated antigen (HMW-MAA) was detected only in fetal skin and in one nipple of 54 normal tissues from adults tested, while the cytoplasmic MAA recognized by the monoclonal antibody 465. 12S was found in most of the normal tissues tested. Among malignant lesions, the HMW-MAA was found in melanomas, astrocytomas, and skin carcinomas; the cytoplasmic MAA was found in all of the malignant lesions tested, even those which originated from normal tissues without detectable cytoplasmic MAA. The levels of the HMW-MAA and of the cytoplasmic MAA showed marked variations in malignant lesions removed from various patients, as well as in autologous metastatic lesions removed from four patients with melanoma. No relationship was found between the degree of expression of the two MAA analyzed and the clinical stage of the disease. Both types of MAA were found in sera from patients with melanoma or other types of cancers, as well as in sera from healthy donors. The level of the HMW-MAA tended to be higher in patients with Stage IV melanoma.
Antigen-Antibody Complex↗
Potential of palladium-109-labeled antimelanoma monoclonal antibody for tumor therapy.
Palladium-109, a beta-emitting radionuclide, was chelated to the monoclonal antibody 225.28S to the high molecular weight antigen associated with human melanoma. The radiolabeled antibody maintained its specific in vitro reactivity with cultured human melanoma cells. Injection of the radiolabeled monoclonal antibody into nude mice bearing human melanoma resulted in significant accumulation of the radiolabel in the tumors: 19% injected dose/g; 38:1 and 61:1 tumor-to-blood ratios at 24 and 48 hr, respectively. The localization of the radiolabeled antibody in liver and kidney also was high, but appreciably lower than that achieved in tumor. These results suggest that Pd-109-labeled monoclonal antibody to tumor-associated antigens may have potential applications in tumor immunotherapy.
Animals↗
Modulation by recombinant DNA leukocyte (alpha) and fibroblast (beta) interferons of the expression and shedding of HLA- and tumor-associated antigens by human melanoma cells.
With a panel of monoclonal antibodies, the effect of recombinant human leukocyte interferons (i.e., IFN-alpha A, IFN-alpha D), of a hybrid leukocyte IFN (i.e., IFN-alpha A/D (Bg1)), and of recombinant fibroblast (beta) IFN on the expression and shedding of four types of melanoma-associated antigens (MAA) and of HLA antigens by the cultured melanoma cell line Colo 38 was investigated. None of the IFN affected the expression of the high m.w. melanoma-associated antigen (HMW-MAA), but all of them increased its shedding. The expression and shedding of the 115,000 MAA and of the 100,000 MAA were increased by IFN; the magnitude of the effect as well as the kinetics were different for the various IFN preparations. The cytoplasmic MAA was the most sensitive to modulation by IFN, because all four types increased its surface expression, its total content, and its shedding. The three types of leukocyte IFN, as well as the fibroblast IFN, were all effective in increasing the expression of HLA-A,B,C antigens, the effect being more marked on the free heavy chain than on the HLA-A,B,C complex. However, only leukocyte IFN enhanced the shedding of the HLA-A,B,C molecular complex. The three types of leukocyte IFN and especially fibroblast IFN enhanced the expression of the gene products of the HLA-D region, the effect being more marked on DC-1 antigens than on HLA-DR antigens. No effect on the shedding of HLA-DR and DC-1 antigens was detected.
Antibodies, Monoclonal↗
Re.: Purification of IgG monoclonal antibody by caprylic acid precipitation.
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Animals↗
Isolation of viable melanoma cells from surgically removed lesions using dishes coated with monoclonal antibody to a high molecular weight melanoma associated antigen.
The panning methodology has been applied to isolate viable human melanoma cells from surgically removed lesions. In this procedure a monocellular suspension mechanically prepared from a biopsy is incubated in plastic dishes coated with the monoclonal antibody (MoAb) 225.28S to a membrane bound high molecular weight-melanoma associated antigen (HMW-MAA). The melanoma nature of the cells growing in MoAb 225.28S coated dishes is indicated by the specific reactivity with anti-HMW-MAA MoAb, by its detection in spent culture medium and by morphological criteria. The specificity of the procedure is proven by the lack of growth of cells seeded in fetal calf serum (FCS) coated dishes, as well as of cells lacking the HMW-MAA in MoAb 225.28S coated dishes. The adherence of melanoma cells to plastic dishes is influenced by the concentration of the plastic bound MoAb 225.28S and by the incubation time. Melanoma cells isolated by adherence to MoAb 225.28S coated plates do not display any detectable change in their growth curve and colony forming ability. The ready availability of melanoma cells isolated from surgically removed lesions will greatly facilitate the characterization of the interaction between host's immune system and tumor cells and the screening of anti-tumor agents in preclinical tests.
Antibodies, Monoclonal↗