Antiseea to Carcinoembryonic Antigen: Removal of Persistant Impurities (1)

Antisera to carcinoembryonic antigen preparations frequently contained antibodies to a distinct normal serum protein(s). The latter antibodies persisted following absorptions with cross-linked human plasma proteins, normal colon, and Group 0 red cells. A simple method for removal of the contaminating antibodies involved absorptions with the perchloric acid-soluble fraction of normal serum. Excesses of the absorbant and its soluble immune complexes were then removed on anion exchange columns.     

Depletion of IgG in rabbit sera by absorption with some group A streptococci.

Streptococci group A, type M 1 or M 56 were capable to deplete IgG in normal rabbit sera during absorption by means of a surface Fc receptor. One absorption with 8.0 mg (dry weight) M 1 streptococci or two absorptions with 71 mg M 56 streptococci removed all IgG in 0.2 ml rabbit serum diluted 1:10 (containing 1.5 mg IgG/ml). Antibodies to egg albumin and antibodies to sheep red cells were not demonstrable after absorption of 0.2 ml serum diluted 1:10 with 16.0 mg M 1 streptococci. These results are discussed in relation to some reports on immunologic cross-reactions between streptococci and mammalian tissues.     

Parallelism of serum anti-F(ab')2 and anti-cationic IgG reactivities in patients with systemic lupus erythematosus

Cationic anti-DNA antibodies may be related to glomerular injury in murine lupus nephritis or in patients with systemic lupus erythematosus (SLE). Therefore, anti-cationic antibodies in SLE could include antibodies with regulatory function on such pathogenic cationic molecules. Since anti-F(ab')2 antibodies may be involved in the idiotype control of anti-DNA antibodies in some patients with inactive SLE, the present study was aimed to determine if SLE patients with significant serum levels of anti-F(ab')2 produce antibodies reacting with cationic IgG molecules. Three SLE sera with high titers of anti-F(ab')2 antibodies were individually adsorbed by sequential affinity chromatography on three Sepharose columns coupling normal IgG from Cohn Fraction II, pooled cationic IgG myeloma paraproteins displaying idiotypic anti-DNA markers (F4 and 8.12), and F(ab')2 fragment from allogeneic IgG, respectively. Eluates obtained from cationic IgG adsorption showed predominant anti-F(ab')2 reactivity. A similar profile was also detected in a serum from a normal control donor with high levels of anti-F(ab')2. Biotinylation of anti-cationic eluates showed that such antibodies were significantly more reactive with cationic than anionic or neutral IgG, confirming their apparent affinity for positively charged antigens on IgG molecules. Since anti-cationic absorptions were able to remove the anti-F(ab')2 activities in the SLE sera studied, it is possible that anti-cationic antibodies could function as immunoregulatory antibodies in the idiotypic control of some SLE autoreactive phenomena, including glomerular anti-DNA deposition.     

Antibody Formation to Embryonic Intestine in Syngeneic Rats

The capacity to produce antibodies to fetal antigens in syngeneic animals has been established. Fetal intestine from inbred Lewis rats injected into adult rats of the same strain provoked fetal intestine-specific antibodies in the serum as judged by immuno-fluorescence. In addition, autoantibodies to intestinal tissue were produced but these were shown by serological absorptions to be distinct from the anti-embryonic activity.     

Partial purification of human kidney antigens.

A chromatographic method using immunoaffinity has been utilized in the search for kidney-specific antigens. After repeated absorptions by liver proteins coupled to a column of Sepharose CN-Br of a rabbit anti-human kidney serum, several antibodies remained. When these antibodies were coupled to a similar column and used for immunoaffinity chromatography, 8 kidney antigens, studied as peptides with molecular weights of 33,000, 80,000, 98,000 and 110,000-125,000 were obtained. They should not be found in similarly treated liver extracts.     

Absorption, Elution and Blocking Studies with the Complex Antiserum ST

Quantitative absorptions and elutions were performed with the broadly reactive serum ST. The major component of this serum is directed towards the B-locus antigens Bw35, B5, and B18, which comprise the 4c CREG. A second component contains lower titered antibodies which react with members of the 4c CREG, but also appear to detect qualitative and quantitative variations associated with B-locus specificities, B15, B17 and B8. Blocking studies indicate that these antibodies do not bind to the same sites on B8 cells as well-defined anti-B8 sera. These findings are discussed in terms of public specificities at the B-locus or a new specificity at a closely linked locus.     

The immunoglobulin-like T-cell receptor. I. In situ demonstration of immunoglobulin Fab-region determinants of rodent T- and B-lymphocytes using chicken antibodies.

Antibodies were raised in chickens to the (Fab')2 fragment of normal murine IgG and to the k-myeloma protein MOPC 41. Following appropriate absorptions or purification by immune affinity chromatography the chicken antibodies bound specifically to both T- and B-cells of mice, rats and guinea-pigs as detected by quantitative cytofluorescence, radioactive binding assays and transmission and scanning immunoelectronmicroscopy. These antibodies are directed against polypeptide determinants of the Fab fragment, block antigen binding by purified idiotype-bearing murine antibodies, and provide useful probes for visualization and analysis of immunoglobulin-like surface receptors of rodent B- and T-cells.     

Suppression of Lymphoblastoid Cell Line Proliferation by Antisera to HLA-DR and Other HLA Antigens

We have observed that certain antisera to HLA antigens suppress the in vitro proliferation of lymphoblastoid cell lines. An antiserum to HLA-B8 demonstrated suppressor activity associated with the presence of B8 antigen on the target LCLs; this activity was removed by absorption with platelets or with B8-positive LCLs. An antiserum to HLA-DRw7 suppressed proliferation of all six DRw7-positive LCLs and none of 12 DRw7-negative LCLs; this activity was removed by absorptions with three DRw7-positive LCLs, each representing a different HLA-D allele (i.e. Dw7, 10 and 11); it could not be removed by triple serial absorptions with platelets from multiple-donor pools. These experiments indicate that the suppressor activity of this serum is specifically associated with antibodies to DRw7. Our model of LCL growth suppression by HLA antisera is easily manipulated and provides a definitive tool for further characterizing HLA antigens and antisera in a complement-independent system.     

THE IMMUNOGLOBULIN-LIKET-CELL RECEPTOR: I. IN SITU DEMONSTRATION OF IMMUNOGLOBULIN Fab-REGION DETERMINANTS OF RODENT T- AND B-LYMPHOCYTES USING CHICKEN ANTIBODIES*

Antibodies were raised in chickens to the (Fab')₂ fragment of normal murine IgG and to the κ-myeloma protein MOPC 41. Following appropriate absorptions or purification by immune affinity chromatography the chicken antibodies bound specifically to both T- and B-cells of mice, rats and guinea-pigs as detected by quantitative cytofluorescence, radioactive binding assays and transmission and scanning immunoelectronmicroscopy. These antibodies are directed against polypeptide determinants of the Fab fragment, block antigen binding by purified idiotype-bearing murine antibodies, and provide useful probes for visualization and analysis of immunoglobulin-like surface receptors of rodent B- and T-cells.     

MLR-blocking antibodies are directed against alloantigens expressed on syncytiotrophoblasts

In this study, the reactivity of mixed lymphocyte reaction-blocking antibodies toward trophoblasts was investigated. Nineteen sera that had a significant level of MLR-blocking effect were obtained from normal pregnant women and from primary habitual aborters after immunotherapy. The blocking effects of these sera were studied before and after absorption with syncytiotrophoblast plasma membrane (STPM) from the respective mothers' placenta (autologous) as an experimental group and from unrelated mothers' placenta (allogeneic) as a control. The HLA types of cord blood lymphocytes corresponding to ten of these STPM were then determined. In the study with the 11 normal pregnancy sera, the blocking effect of every serum was partially removed by autologous STPM absorption. This revealed a significant difference between the preabsorption and postabsorption MLR-blocking effect. Nevertheless, absorption by the allogeneic STPM either partially removed or did not remove the blocking effect. Furthermore, there was a statistically significant difference in the elimination of the MLR-blocking effect between the autologous STPM and the allogeneic STPM. Studies with the sera from the immunized aborters revealed similar results. There was no statistically significant difference in the elimination of the antibodies between absorption with autologous and with allogeneic STPM. In these habitual aborters' sera, comparison of paternal stimulator and cord blood lymphocytes HLA types showed that the positive allogeneic STPM absorptions could not be explained sufficiently by coincidence of HLA types. Further studies are needed to determine whether the alloantigens are HLA-linked or not.     

Cross-reactivity distinguishes serum and nephritogenic anti-DNA antibodies in human lupus from their natural counterparts in normal serum

To distinguish the properties of anti-DNA antibodies in patients with lupus from those in normal individuals, we compared the ligand binding, idiotypic and charge properties of serum anti-DNA antibodies derived from: patients with active lupus; normal individuals; and among Ig eluted from the kidneys of two patients with active lupus nephritis (one with mesangial proliferation and the other with membranous nephropathy). The kidney eluate anti-DNA antibodies were the most cross-reactive; they cross-reacted with ssDNA, poly(GdC), poly(dT), poly(dG), poly(dC), ZDNA, SmRNP and the phospholipids cardiolipin and phosphatidyl serine. Lupus serum anti-DNA antibody cross-reacted with polynucleotides but not with phospholipids, whereas anti-DNA antibodies derived from normal serum reacted only with poly(dT). An anti-idiotype (anti-IdD; produced against serum anti-DNA antibodies from one patient) reacted with: anti-DNA antibodies in 8/9 lupus sera; antibodies in both kidney eluates; and anti-DNA antibodies from 5/7 normal sera. Anti-IdD did not react with Ig that did not bind to DNA. Isoelectric focusing of Ig showed that the charge of anti-DNA antibodies from lupus serum and normal serum were similar and unrestricted (pI 5.4-9.0); Ig in kidney eluates varied: membranous lupus pI 4.5-8.6; mesangial lupus pI 8.1-9.1. We conclude that idiotypically related anti-DNA antibodies in tissue lesions, lupus serum and normal serum from different individuals can be distinguished on the basis of their cross-reactive antigen-binding properties. Furthermore the cross-reactive properties of lupus auto-antibodies may influence their capacity to form glomerular immune deposits.     

Unique antigenic determinants (idiotypes) used as markers in a patient with macroglobulinemia and urticaria. Similar idiotypes demonstrated in the skin and on peripheral blood lymphocytes

An antiserum was raised against a monoclonal IgMk macroglobulin isolated from serum of a patient with recurrent urticaria. The antiserum was made idiotype-specific through adequate absorptions. The anti-idiotype antiserum reacted only with the immunizing protein and its Fab fragments and not with other monoclonal proteins of IgM and IgA class or pooled IgG as assayed in an enzyme-linked immunosorbent assay. IgM antibodies with the same idiotype as the monoclonal IgM protein were detected in the dermal/epidermal junction area of diseased skin. The similar idiotypic determinants could also be demonstrated on membrane-bound molecules of peripheral blood B and T lymphocytes using the immunofluorescence methods.     

Functional analysis of T and B cells from blood and thyroid tissue in Hashimoto''s disease

The existence of antibodies against alcohol-treated rabbit hepatocytes in sera of patients with alcoholic liver disease was investigated utilizing a 125I-labelled protein A assay. The sera, after two absorptions with normal rabbit hepatocytes, were incubated with hepatocytes isolated from rabbits which had been treated for 4 days with a daily dose of ethanol (1 g/kg body weight) intravenously. Serum antibodies against alcohol-treated hepatocytes were detected in 21 of 55 patients with alcoholic liver disease; this was associated with a distinct, mixed granular and linear fluorescence staining the surface of alcohol-treated hepatocytes. By contrast, none of 23 patients with other liver diseases who had no history of excessive alcohol intake showed the antibodies in their sera. The values of the antibodies of patients with alcoholic active cirrhosis and alcoholic hepatitis were significantly higher than those of other types of alcoholic liver disease. These results showed that antibodies against alcohol-treated hepatocytes are present in alcoholic liver disease with inflammation. The role of these antibodies on liver cell damage in alcoholic liver disease remains to be clarified.     

The neutralization of pseudorabies virus by anti-alpha-galactocyl natural antibody in normal serum

Pseudorabies virus (PrV), a member of herpesviridae alphaherpes subfamily, can infect human cells in vitro. However, the transmission to Old World primates including humans is strongly restricted. In this study, we report the neutralizing activity of normal human serum against PrV grown in CPK cells derived from pig. PrV grown in all Old World primates-derived cells, which was tested in this study, were not neutralized by normal human serum. The virion of PrV grown in CPK cells harbored Galalpha1-3Galbeta1-4GlcNAc-R (alpha-gal epitope) on its surface, while PrV grown in Vero cells did not. Depletion of antibodies reacting to surface antigens of CPK cells negated the neutralization activity of human serum. Blockade of anti-alpha-gal antibodies by adding soluble Galalpha1-3Gal to normal human serum also prevent the inactivation of PrV grown in CPK cells. Although normal swine serum did not neutralize PrV grown in CPK cells, swine serum supplemented with exogenous anti-alpha-gal antibodies did. These results indicate that anti-alpha-gal antibodies in normal human serum contribute to the neutralization of PrV. Anti-alpha-gal antibodies in normal human serum may prevent transmission of PrV into humans.     

Human B-lymphocyte heteroantigens: direct evidence for specificities not shared by monocytes

A heterologous anti-human B-lymphocyte serum was raised in the rabbit and rendered specific by extensive absorptions. After absorptions with erythrocytes and thymocytes, the antiserum was cytotoxic for an average of 18·5% of normal peripheral blood lymphocytes. Most chronic lymphatic leukaemia cells and `hairy cells' were sensitive to this antiserum but `null cells' were not. E-rosette-forming cells were insensitive to the antiserum, while EAC-rosettes and rosettes with mouse erythrocytes were partially or totally inhibited. In ultrastructural characterization studies after immunoperoxydase labelling, it was demonstrated that not only B lymphocytes, but also virtually all monocytes having phagocytized latex particles, were stained. A further absorption with monocytic cells from a patient with subacute myelomonocytic leukaemia was thus performed and the resulting antiserum was then specific for human B lymphocytes only and did not label any latex-phagocytizing monocyte. These results demonstrate that besides antigens common to monocytes, B lymphocytes express specificities characteristic of the B-cell population only, at their surface.     

Antigenic difference between intracellular and membrane antigens induced by herpesvirus of turkeys

Summary The difference between intracellular antigen (IA) and membrane antigen (MA) induced by herpesvirus of turkeys (HVT) was examined by immunofluorescence analysis. Convalescent sera from chickens infected with HVT (HVT convalescent serum) and hyperimmune sera from chickens immunized with partially purified virus (anti-HVT particle serum) or with a major HVT soluble precipitin antigen (anti-common Ag serum) were used as the source of antisera to specify both antigens in these experiments.Absorption of a convalescent serum from HVT-inoculated chicken with IA positive cells, resulted in reduction of their reactivity to IA but not to MA. Differential absorption of the hyperimmune sera by IA positive cells and MA positive cells resulted in reduction of their reactivity to homologous cells but not to heterologous cells which were used in the absorptions. When the relationship between the levels of anti-IA and anti-MA antibodies was examined by a blocking immunofluorescence test using 38 sera from chickens inoculated with HVT, no correlation was observed (p>0.25). These results provide evidence that MA and IA are antigenically different from each other.With 3 Figures     

Tumour cell-antibody interactions. I. In vivo experiments.

Goats were immunized with membrane fractions of the L5178Y tumour syngenic to DBA/2 mice. IgG fractions of this antiserum were made tumour specific by repeated in vitro and in vivo absorptions with normal cells and concentrated by adsorption to and elution from tumour cells. In vivo studies indicated that the accumulation of 125I-labelled antibody in tumour tissue could not be improved by extensive purification and concentration. The observed clearance rates of radioactivity from tumours and whole animals showed that the metabolism of antibodies was significantly accelerated in the presence of target cells. It is suggested that a rapid neutralization and degradation of antibody by the target tissue prevents its accumulation in tumour nodules.     

IgG4 and release of histamine from human peripheral blood leukocytes

Allergen-specific IgG4 was found in the sera from many normal, nonallergic individuals. Basophil leukocytes from donors with IgG4 antibodies to an allergen, but without IgE antibodies to the same allergen, did not release histamine when challenged with this allergen. In some cases, anti-IgG4 antiserum induced a release of histamine from the leukocytes. However, these cells also release histamine after incubation with normal rabbit serum or normal sheep serum. It is concluded that the IgG4-RAST cannot be used for the detection of IgG short-term sensitizing antibodies.     

An improved enzyme-linked immunosorbent assay of anti-collagen antibodies in human serum

An improved enzyme-linked immunosorbent assay (ELISA) for the determination of anti-collagen antibodies in human serum has been developed. The method is based on the use of serum samples diluted to 1/50 with heat-inactivated normal rabbit serum adjusted to pH 8.0 with solid Tris (0.05 M), NaCl (0.15 M) and 2 M HCl. The use of normal rabbit serum minimizes non-specific adsorption of immunoglobulin G onto the plastic surface of microtiter plate. The applicability of the method for the quantitation of anti-collagen antibodies in human serum is demonstrated with 290 specimens of sera from normal controls (194) and patients with rheumatoid arthritis (96).     

68例糖尿病肾病患者血清超敏C反应蛋白、血清抗氧化型低密度脂蛋白抗体检测结果分析

目的:探讨糖尿病肾病与超敏c反应蛋白(hs-CRP)、抗氧化型低密度脂蛋白(OX-LDL)抗体的关系。方法:测定68例糖尿病肾病患者及50例正常对照组血清超敏C反应蛋白、血清抗氧化型低密度脂蛋白抗体,并将两组结果作对比分析。结果:糖尿病肾病患者血清hs-CRP、血清OX-LDL抗体均高于正常对照组,两组之间差异具有统计学意义(P<0.01)。结论:血清hs-CRP、血清OX-LDL抗体在糖尿病肾病组中明显升高,可作为糖尿病肾病发生、发展的重要指标。