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Biomedical subjects

R D Rossen

Publications and source records attributed to R D Rossen.

At least 55 records · Page 3Linked to original sources
PubMed1984

Regulation of antibody release by naturally occurring anti-immunoglobulins in cultures of pokeweed mitogen-stimulated human peripheral blood lymphocytes.

Immunoregulatory influences of human anti-immunoglobulins (anti-Ig) were studied in cultures of peripheral blood lymphocytes (PBL) from 11 normal donors. Pokeweed mitogen (PWM)-stimulated PBL released anti-Ig specific for Fab or Fc fragments of IgG, often within the first 24 to 72 hr in vitro. PBL that released more than 1 ng/ml IgM anti-Fab during the first 72 hr in vitro ultimately produced significantly less antibody (Ab) by the 12th day than PBL that released no detectable IgM anti-Fab during the first 3 days in culture. Adding affinity-purified human anti-Fab to PWM-stimulated PBL also suppressed the later Ab release by these cells. Suppression was polyclonal, affecting IgM anti-Fc, IgM anti-Fab, and IgM anti-tetanus toxoid Ab, and was directly dependent on the quantity of anti-Fab added. Anti-Fab Ab, isolated from single donor sera, were more suppressive, nanogram for nanogram, than were equal quantities of IgG anti-Fab obtained from Cohn Fraction II, when added to autologous donor PBL in vitro. Affinity-purified IgM anti-Fc, from pooled rheumatoid arthritis patient sera, also suppressed Ab release by PWM-stimulated PBL in a dose-dependent manner. These observations suggest that anti-Ig may exert a significant immunoregulatory role in man that can override to some extent the T cell-dependent stimulus for polyclonal B cell activation provided by PWM.

Antibodies, Anti-Idiotypic↗
PubMed1984

Effects of gamma-irradiation on chromosomes of cultured lymphocytes from rheumatoid arthritis patients and their family members.

Increased rates of spontaneous or induced chromosome breakage are seen in many types of diseases, including the 'collagen-type' autoimmune diseases. Using a 60Co gamma cell, we irradiated lymphocyte cultures from three related rheumatoid arthritis patients, their immediate family members, and two unrelated rheumatoid arthritis patients. Although these individuals had not shown abnormally high levels of spontaneous chromosome breakage, they did show an abnormal sensitivity to irradiation, which was manifested in several ways. Two of the probands showed induced breakage rates that were twice as high as those seen in controls. In addition, the reduction of mitotic index, due either to increased cell death or to induction of a G2 lag period, was higher in the arthritis group (including non-symptomatic family members) than in the control group. Finally, we observed a high frequency of an unusual type of cell in the arthritis group. These unusual cells resembled c-anaphases seen with extended colcemid treatment, and may indicate that the mitotic apparatus in cells from this group is particularly sensitive to ionizing radiation.

Arthritis, Rheumatoid↗
PubMed1983

Use of human antibodies to identify antigens in cultured human tumor cells: detection of discrete antigen molecules using electroblotting and enzyme-linked antibody probes.

To demonstrate antigens in cultured human tumor cells recognized by antibodies in human sera, extracts of cultured malignant melanoma cells were separated by SDS-PAGE, and electrophoretically transferred to nitrocellulose paper (NCP). The resulting electroblots were incubated with human serum, and bound immunoglobulin (Ig) was visualized with rabbit antibodies specific for human IgG, IgA or IgM, followed by peroxidase-conjugated goat anti-rabbit IgG. Antigen-antibody reactions in the nitrocellulose paper were also detected using 125I-labeled anti-rabbit IgG. As little as 125 pg of bound antibody per band were detectable. The numbers of proteins recognized by antibodies in human sera depended both on the quantity of protein transferred and the concentration of Ig applied to the NCP. Whole serum could not be used at dilutions less than or equal to 1:20 without an unacceptable increase in background staining. Binding of Ig to tumor cell proteins transferred to NCP depended on interactions with the Fab', not the Fc region of the Ig molecule. To determine the efficiency of transfer as a function of both time and molecular weight, tumor cell proteins were intrinsically labeled with 75Se-labeled methionine and transferred for up to 4 h after fractionation in gels containing acrylamide concentrations of 5%, 7.5%, 10% or 12%. Proteins less than 150 kDa were transferred with particularly high efficiency in greater than or equal to 2 h. Different antigens were recognized by the IgA, the IgG and the IgM molecules from the same sera. The methods outlined herein are proving to be useful in monitoring the purification of specific antigens from whole tumor cell extracts.

Antibodies, Neoplasm↗
PubMed1983

Anti-Fab' antibodies in rheumatoid arthritis. Measurement of the relative quantities incorporated in soluble immune complexes in sera and supernatants from cultured peripheral blood lymphocytes.

High levels of anti-immunoglobulins that react with Fab' fragments of IgG have been observed in sera of patients with rheumatoid arthritis (RA). To determine how much of these anti-Fab' antibodies are incorporated within immune complexes (IC), we added 125I-Fab' to sera and then measured the amount of labeled Fab' that could be precipitated by adding polyethylene glycol (PEG). Sera were either maintained at neutral pH during this procedure or acidified (pH 3) to dissociate IC. Acidification permitted the 125I-Fab' an equal chance to compete with other endogenous antigens for anti-Fab' antibodies, once excess hydrogen ion was removed. Quantities of anti-Fab' in sera of 20 seropositive RA patients were greater than in sera of 43 age- and sex-matched healthy controls, as measured by this assay and by a solid phase radioimmunoassay. However, significantly less anti-Fab' antibody was incorporated in ICs in the RA patients' sera. Supernatants from cultured peripheral blood lymphocytes of RA patients also contained relatively more "free" and less "hidden" anti-Fab' than culture supernatants from controls. Thus there appear to be qualitative as well as quantitative differences in the anti-Fab' antibodies synthesized by RA patients. This may reflect different proportions of IgM and IgG anti-Fab' in their sera, differences in the average avidity of these antibodies, or differences in the reciprocal relationships within the idiotypic network that result in release of antibodies by certain antibody-producing cells, but not by clones that produce complementary idiotypes.

Adult↗
PubMed1983

Correlation of immune complexes in disseminated neuroblastoma with serum antibody to bovine serum albumin.

To evaluate the relationship between tumor burden and circulating immune complexes (IC) in children with neuroblastoma (NBL), we studied sera collected at intervals from patients with disseminated (Stage III or IV) NBL. Sera from 10 of 12 patients contained IC by the Raji cell assay at some time during the first 9 to 11 months of the study. Higher IC levels were observed in sera of female patients. Fluid-phase C1q binding tests detected IC in only 16% of sera. IC measurements by either assay did not correlate with tumor burden. However, serum IC levels, as measured by the Raji cell assay, correlated significantly with serum antibody to bovine serum albumin (BSA) (rs = 0.54; p less than 0.001, rs = r as determined by Spearman rank correlation test). Measurement of anti-BSA antibodies in sera from the 12 patients, tested serially for circulating IC, and from five additional patients revealed that these had significantly higher anti-BSA activity than was found in sera from 13 age-matched controls. Sera from females also had relatively high levels of anti-BSA. Levels of antibody to bovine gamma-globulin and casein were not abnormal. Three sera with high IC levels (greater than 800 micrograms equivalents of heat-aggregated IgG) and relatively low anti-BSA activity appeared to contain "hidden" antibodies to BSA. These were demonstrated by measuring the increase in the ability of sera to bind 125I-BSA after they had been briefly acidified and then neutralized in the presence of the labeled BSA. The possible relevance of these results to the pathophysiology of NBL is discussed in light of earlier work that reported that serum IC levels correlate with the stage of the disease in NBL.

Animals↗
PubMed1983

Circulating immune complexes and tumor cell cytotoxins as prognostic indicators in malignant melanoma: a prospective study of 53 patients.

To evaluate the relationship between tumor burden and circulating immune complexes (IC) in malignant melanoma, we tested sera collected serially from 15 normal donors and 53 patients. Forty-eight of these had Stage III or IV disease at the outset of the study. The median survival time (MST) of ten patients with Stage IV disease whose sera contained C1q-binding IC at the outset of the study was 4.7 months; the MST of the 25 Stage IV patients whose sera were initially free of IC by this test was 8.65 months (p less than 0.02). C1q-binding IC were not found in the initial serum samples from 13 patients with Stage III or 5 patients with Stage I disease. Abnormal C1q binding tests were measured in 4 of 67 sera (6%) from 13 patients who remained free of evident tumor for up to 41 months. IC were detected in 13 of 39 sera (33%) from 19 patients with progressively growing tumors and in 21 of 68 sera (31%) from 21 patients who were initially free of disease but developed recurrences later, or who had significant remissions of variable duration during follow-up. The MST of 31 patients whose serial serum samples remained free of C1q-binding IC was 15.8 months. Twelve patients whose sera were initially free of circulating IC later developed abnormal serum C1q-binding levels. Their MST was 10.3 months. The MST of ten patients with persistently abnormal serum IC levels was 4.7 months. C1q-binding IC were reciprocally related to the presence of complement-dependent antibodies, cytotoxic for cultured allogeneic malignant melanoma cells in sera from 29 of these patients (r = -0.491;p = 0.003). These results suggest that the appearance of circulating C1q-binding IC is pathophysiologically important in malignant melanoma. Measurement of C1q-binding IC may be useful in assigning prognosis in this disease.

Antigen-Antibody Complex↗
PubMed1983

Characterization of anti-Fab' antibodies in human sera: identification of soluble immune complexes that contain hidden anti-KLH and blocking anti-immunoglobulins following immunization with keyhole limpet haemocyanin.

After immunization with keyhole limpet haemocyanin (KLH), increased concentrations of anti-KLH and anti-Fab' antibodies (Abs) were demonstrated in sera from 18 of 20 volunteers. In many cases, post-immunization sera contained soluble immune complexes that incorporated both anti-Fab' and 'hidden' or 'blocked' anti-KLH antibodies. The complexes containing hidden anti-KLH and blocking anti-Fab' Abs were not found in pre-immunization sera. The hidden Abs to KLH were revealed by demonstrating increases in anti-KLH activity in sera incubated previously with Fab' fragments, immobilized on plastic microtitre tray wells. Incubation with insoluble Fab' did not influence the quantity of anti-tetanus toxoid (TT) that was detected in these sera. Addition of affinity purified anti-Fab' Abs to samples, previously 'unblocked' by adsorption with immobilized Fab', depressed their anti-KLH activity to levels present before adsorption, but did not change the quantity of Abs to Dermatophytin, Trychophyton, or TT therein. These results suggest that some autoantibodies generically recognized as 'Fab' specific' have properties that are usually considered to be characteristic of autoanti-idiotypes.

Adult↗
PubMed1983

Synthesis of antigens, cross-reactive with bovine serum albumin, by cultured neuroblastoma cells.

Antisera to bovine serum albumin (BSA) react with biosynthetic products of the LAN-1 neuroblastoma cell line. This immunological reaction was shown by analysis of immunoprecipitates prepared with anti-BSA and products of this cell line intrinsically labeled by the incorporation of 3H- or 35S-labeled amino acids. Moreover, antibodies in sera from two patients with neuroblastoma precipitate intrinsically labeled macromolecules produced by these tumor cells. Release of antigens cross-reactive with BSA by neuroblastomas may explain, in part, the high levels of antibody to BSA and circulating immune complexes containing hidden or "blocked" antibodies to BSA found in some patients with this tumor.

Antigens↗
PubMed1982

Thrombotic thrombocytopenic purpura complicating Legionnaires' disease.

A case of Legionella pneumophila infection complicated by thrombotic thrombocytopenic purpura (TTP) was confirmed at autopsy by the demonstration of the organism in lung tissue, and by the finding of widespread intravascular and subendothelial thrombi associated with microinfarctions in all major organs examined. In addition to the typical hematologic abnormalities of TTP, the patient was found to have a low serum C3 level and elevated levels of immune complexes as measured by the liquid phase C1q binding assay. We suggest that the L pneumophilia infection caused endothelial damage and/or platelet aggregation, perhaps as a consequence of complement activation, thus initiating the sequence of events leading to extensive microvascular thromboses.

Antigen-Antibody Complex↗
PubMed1982

Familial rheumatoid arthritis: a kindred identified through a proband with seronegative juvenile arthritis includes members with seropositive, adult-onset disease.

Segregation of chromosome #6 markers has been studied in a large family, identified by a proband with seronegative, juvenile-onset rheumatoid arthritis, which contains four other individuals with adult-onset rheumatoid arthritis (RA). Two of the adult-onset patients have classical seropositive RA. Sera from two healthy members of this family also contain rheumatoid factors (RF). Six family members had crossovers in the short arm of chromosome #6. Three individuals with recombinants between HLA-B and HLA-D were identified; three others had identifiable crossovers between HLA-D and GLO (Glyoxylase 1). Linkage analysis suggested that susceptibility to RA in this family was influenced by a dominant gene located centromeric to HLA-B. The highest lod score (1.64) was obtained for linkage to GLO at a recombination rate of zero. Inheritance of specific chromosome #6 or Gm immunoglobulin allotype markers did not appear to influence serum RF. These results agree with previous family studies which suggest that acquisition of childhood- and adult-onset RA is influenced by a common disease susceptibility gene, linked to the major histocompatibility gene complex.

Adolescent↗
PubMed1982

Production of antibodies specific for Fc, Fab', and streptokinase-streptodornase in vitro by peripheral blood cells from patients with rheumatoid arthritis and normal donors. Identification of immune complexes in culture supernatants containing hidden antibodies reactive with Fab' fragments of immunoglobulin G.

To study antibody (Ab) biosynthesis in rheumatoid arthritis (RA), the immunoglobulin (Ig)M anti-Fc, anti-Fab', and antistreptokinase-streptodornase (SKSD) produced by peripheral blood lymphocytes (PBL) were measured at intervals from 1 to 19 d in culture. PBL from 17 seropositive patients with active RA and 30 age-matched controls were evaluated. Within the first 24 h, PBL from six of eight patients released >30 ng IgM anti-Fc, even in the absence of pokeweed mitogen (PWM). This early release of Ab was blocked by cycloheximide. With or without PWM, PBL from normal donors did not release IgM anti-Fc until after 3-5 d in vitro. By day 9, unstimulated PBL from seven patients made > 100 ng IgM anti-Fc. Un-stimulated PBL from normals never made >95 ng of this Ab. When PWM was added, PBL from normal donors released as much IgM anti-Fc as was found in RA donor cultures. Paradoxically, addition of PWM to PBL of RA patients suppressed release of IgM anti-Fc in 4 of 17 cases to levels significantly below those found in unstimulated cultures of the same cells. Without PWM, PBL from RA donors frequently failed to make IgM anti-SKSD (P < 0.05 compared with normal donors' cells). With PWM, the quantities of IgM anti-SKSD released were comparable. Fluctuations in IgM anti-Fab' levels during the life-time of these cultures were sufficient to suggest that these Ab may be taken up in immune complexes. This hypothesis was verified by acidifying (pH 3.1) culture supernatants to which (125)I-Fab' had been added previously. The samples were then neutralized (pH 7.6) and 12% polyethylene glycol was added to separate free from antibody-bound (125)I-Fab'. This procedure increased the quantity of (125)I-Fab' precipitated by > 10-fold in some cases. These studies suggest that there are a variety of abnormalities in Ab biosynthesis in RA. These include spontaneous synthesis of comparatively large quantities of IgM anti-Fc, relatively suppressed release of IgM anti-SKSD, and a paradoxical reduction, in some cases, in the biosynthesis of IgM anti-Fc after addition of PWM.

Antibodies, Anti-Idiotypic↗
PubMed1982

"Hidden" cytotoxic antibodies that react with allogeneic cultured fetal and tumor cells contained in soluble immune complexes from normal human sera.

Hidden complement-dependent cytotoxins were demonstrated in normal donor sera after removing anti-immunoglobulins that normally block the ability of these immunoglobulin Gs to react with surface antigens on tumor cells. The blocking antibodies had certain properties of anti-idiotypes. Immunoglobulin G from Cohn Fraction II, after removing these anti-immunoglobulins, was cytotoxic for cultured fetal cells and for malignant melanoma, breast, lung, colon, and other tumor cells maintained either in tissue culture or by serial passage in nude mice. The cytotoxins were not adsorbed by extracts from normal lymphoid, liver, skin, or red blood cells. These results suggest that a heterogeneous group of natural antibodies reactive with antigens expressed on a variety of neoplastic and fetal cells circulate in normal donor sera as part of a soluble immune complex, together with a blocking anti-immunoglobulin.

Animals↗
PubMed1981

Plasma exchange in selected patients with juvenile rheumatoid arthritis.

Plasma exchange with either fresh-frozen plasma or 5% albumin solution as replacement fluid was performed in four selected patients with juvenile rheumatoid arthritis unresponsive to standard therapy. One 13-year-old boy with life-threatening systemic disease experienced a partial remission of disease and tolerated a decrease in prednisone dose from 15 to 4 mg daily following 14 exchanges with FFP. A 14-year-old girl, dwarfed by systemic disease and long-term corticosteroid therapy, was able to discontinue prednisone and grew 6.3 cm in 11 months following 18 plasma exchanges with FFP. An 8-year-old girl with pauciarticular disease, antinuclear antibody, and uncontrollable iridocyclitis underwent 16 plasma exchanges with 5% albumin solution as replacement; despite removal of antinuclear antibody, her eye disease and arthritis were not helped. A 16-year-old girl with erosive, polyarticular JRA showed no detectable change in her articular disease following nine exchanges. Transient decreases in hematocrit, complement components, and immunoglobulin concentrations occurred. In three patients Westergren sedimentation rate decreased for up to five months after exchanges. One patient died suddenly during an exchange with FFP; the cause of death appeared related to microemboli of unknown nature found in the lungs at autopsy. Plasma exchange should be done only in an intensive care setting and as a research procedure for children with JRA.

Adolescent↗
PubMed1981

Advantages and limitations of staphylococcal protein A-Sepharose for isolating soluble immune complexes from goat, rabbit and human sera.

Staphylococcal protein A (SPA), bound to CNBr-activated Sepharose, was evaluated as a selective adsorbent for soluble immune complexes (ICs) prepared in antigen (Ag) excess. Goat antibody (Ab) to human serum albumin (HSA) and rabbit and human antisera to diphtheria toxoid (DT) were utilized for complex formation. Monomeric goat IgG did not bind SPA. However, HSA-goat anti-HSA complexes which were greater than 12S by sucrose density-gradient ultracentrifugation and had molar Ab:Ag ratios greater than 1.5 were adsorbed, and could subsequently be eluted with acidic phosphate-buffered saline, pH less than or equal to 3.8. Elution with 3.5 M MgCl2 enhanced recovery, but also resulted in hydrolysis of the bound Ab. Ninety per cent of the DT-anti-DT ICs prepared with rabbit Ab and 55% of those prepared with human Ab, in the presence of excess free Ag, bound to the SPA columns. However, only 42% of the DT-rabbit anti-DT complexes, and 32% of those prepared with human antisera were isolated in the acidic phosphate-buffered eluate, free of contaminating proteins. Recovery of ICs by SPA affinity chromatography was significantly decreased when the ICs were partially purified by PEG or ammonium sulphate precipitation before application to the SPA-Sepharose columns. These studies indicate that SPA can be used to isolate ICs prepared in far Ag excess and with Abs which, by themselves, do not bind to this absorbent. They also demonstrate that recovery of ICs from sera using this adsorbent is invariably incomplete.

Ammonium Sulfate↗
PubMed1980

Familial rheumatoid arthritis: linkage of HLA to disease susceptibility locus in four families where proband presented with juvenile rheumatoid arthritis.

The occurrence of a chronic seronegative polyarthritis has been studied in four families in which the proband presented with some form of juvenile rheumatoid arthritis. In these families, histocompatibility testing suggested that susceptibility to arthritis was controlled by a dominant allele with variable penetrance and expressivity at the rheumatoid-like arthritis, first locus (RLA-1). The combined lod scores for the four families (2.70) indicated that the odds in favor of genetic linkage between the major histocompatibility complex and the postulated disease susceptibility gene, RLA-1, were 500:1. In one family, a recombinant event permitted localization of RLA-1 centromeric to HLA-D. Of major interest was the fact that there was significant pleomorphism in the clinical manifestations of arthritis in affected individuals. In some, symptoms first occurred in childhood and in others, in adult life. Even among those with childhood-onset arthritis, different types of juvenile rheumatoid arthritis were observed within the same family.

Adolescent↗
PubMed1979

Circulating immune complexes in colon cancer patient sera.

Sera from 29 colon cancer patients and 16 elderly normal controls were compared using the Raji radioimmunoassay and the Clq binding assay. By the Raji assay 11 of 29 cancer patients had evidence of circulating complexes, while by the Clq assay only 4 of the 29 patients had complexes. There was no significant difference between the levels of circulating complexes in the patient or control groups. With respect to individual patients the results obtained by the Raji assay did not correlate with those obtained by the Clq binding assay.

Adult↗