Antigens in immune complexes from patients with breast cancer

Summary Sera and effusion fluids of patients with breast cancer (BC) contain immune complexes (IC). Antigens present in these complexes were isolated as follows: a pool of effusions from patients with BC was fractionated with ammonium sulfate. The proteins precipitating at 40% saturation were further fractionated by filtration through a Sephadex G-200 column. The material recovered in the first peak (molecules larger than monomeric IgG) was brought to pH 3.0 to dissociate the IC, and the mixture was filtered through a column of Sephacryl S-300 at pH 3.0. Proteins smaller than monomeric IgG were collected, radioiodinated, and used as antigens (¹²⁵Ag) to search for corresponding antibodies in sera of patients with BC (BCS) and of healthy individuals (NHS). ¹²⁵Ag was reacted with the sera and the immune complexes obtained were precipitated with an antiserum to human Ig and analyzed by SDS-polyacrylamide gel electrophoresis followed by autoradiography. Both NHS and BCS contained antibodies against two antigens; one of these appeared as a strong band of 17KD, the other as a doublet of approximately 25KD. It is concluded that some of the proteins in the IC from patients with BC are auto-antigens. No BC-specific antigens were identified.     

Stage-associated incidence of serum circulating immune complexes in patients with untreated bronchogenic carcinoma

Summary Serum circulating immune complexes were quantitated by means of a C1q-binding enzyme-linked immunosorbent assay in the serum from 46 untreated bronchogenic carcinoma patients, and the results compared with those obtained in 48 patients with nonmalignant thoracic diseases and 75 normal healthy donors. The incidence and levels of serum immune complexes in the bronchogenic carcinoma patients were found to be akin to those previously observed, and no modifications of their levels were found to result from surgery. We also found a high degree of association between the presence of immune complexes and the lung cancer stage as defined by the tumor (T), node (N), metastasis (M) system. However, our data indicate that their occurrence has neither a prognostic value, as determined after analyzing the late outcome of the patients, nor a diagnostic one, since the incidence of immune complexes in patients with nonmalignant thoracic diseases was found to be similar to that in the bronchogenic carcinoma patients.     

Activity of antibodies recovered from immune complexes of ovarian cancer patients

Summary Ascites fluids from ovarian cancer patients contain immune complexes (IC). Attempts were made to characterize those antigens to which the patient is reacting using antibody recovered from these complexes. Polyethylene glycol (PEG) precipitation and protein A Sepharose 4B affinity chromatography were used to isolate IC. Dissociation of the IC was achieved by G-150 gel filtration in 0.1 M acetic acid, 0.15 M NaCl. Activity of antibody preparations was measured using a binding assay with ¹²⁵I-labeled staphylococcal protein A. Confluent monolayers of various cell lines (including human ovarian and cervical carcinoma lines, human lymphoblastoid cell lines, human and chick embryo fibroblasts) were used. Six of nine antibody preparations isolated from ovarian cancer patient IC contained at least some reactivity against all cell types.Specificity was further defined using cell adsorption assays and polyacrylamide gel electrophoresis. These results indicate what appears to be autoreactivity directed against a normal component(s) of many cells. No evidence for an ovarian cancer-restricted response was shown. Immunoprecipitation analyses showed several polypeptide bands on autoradiograms (49K, 46K, 33K, 25K), which deviated distinctly from the pattern obtained for whole cell extracts.     

Gangliosides and antibodies to gangliosides in blood serum]

The concentration and composition of gangliosides from normal and pathological blood serum of animals and man are reviewed. Data concerning the elevation of the ganglioside content in the serum under malignization are summarized. The appearance of ganglioside-specific antibodies in some pathological states is described. The possible influence of changes in the serum ganglioside content and composition on immunity is discussed.     

Isolation and dissociation of immune complexes from pleural effusions of lymphoma patients

Summary Immune complexes (IC) isolated from pleural effusions of lymphomas with favorable and unfavorable prognoses were of IgG type. These IC were further dissociated by ion exchange chromatography using 8 M urea. The antibody was found to be a high molecular weight protein (1.5×10⁵ daltons) and reacted with antihuman IgG immunologically while a second peak obtained on ion exchange chromatography may be an antigen moiety with a molecular weight of 3.2×10⁴ daltons as it reacted immunologically with the antibody. Strong cytoplasmic fluorescence was observed with various cell suspensions of lymphomas when reacted with the antibody preparations. The antisera raised against two different antigen fractions prepared from two lymphomas — nHL and LL showed positive fluorescence with both nHL and LL suspensions. The absorption of these rabbit antibodies with individual cell extracts or with antigen preparations also entirely blocked the cytoplasmic staining. The antigen moiety (PK-II) may have a common origin in the disease process. Pleural effusions from patients with unfavorable and favorable prognoses showed identical patterns of separation of IC components.     

Gangliosides from human placenta

Gangliosides of human placenta were studied, using biochemical methods and specific antibodies. The placenta was found to contain three types of gangliosides with oligosaccharide chains Lac, GgOse4 and nLcOse4.     

Circulating immune complexes in patients treated with Pollinex vaccine

Twenty six patients with rhinitis were desensitized with Pollinex. Circulating immune complexes were determined in these patients before and after immunotherapy. Two assay techniques - precipitation and PEG-CCT were applied to assay circulating immune complexes. It was found that specific immunotherapy with Pollinex does not produce circulating immune complexes in patients with hay fever.     

Circulating immune complexes in patients with breast cancer.

In a retrospective study in women with breast cancer circulating immune complex levels were measured by radioimmunoprecipitation with 125I-Clq. Before operation all the patients showed plasma immune complex levels significantly higher than those in controls. Twelve months after mastectomy patients identified clinicopathologically as having a good prognosis had almost normal levels of immune complexes. By contrast, patients with detectable dissemination on diagnosis or those who died within 22 months after mastectomy had significantly raised plasma levels. The tumour-specific nature of the immune complexes detected remains to be shown and suggestions about the applicability of this test not only for prognosis but also for monitoring the course of malignant diseases need to be confirmed by further investigations.     

Gangliosides stimulate astroglial cell proliferation in the absence of serum

Previous studies with microcultures of astroglial (AG) cells from newborn rat cerebrum had shown an ability of gangliosides to interact with AG cells cultured under defined conditions. We have now investigated the capability of gangliosides to stimulate DNA synthesis and cell number increases in similar secondary microcultures of newborn rat cerebrum AG cells. At a concentration of 6 X 10(-5)M, GM1 ganglioside stimulated DNA synthesis and increased cell numbers, with DNA synthesis leading cell increases by 12-24 hr. The ganglioside-induced AG cell proliferative response occurred with GD1a, GD1b and GT1b, GT1b being the most potent at 10(-5)M--while asialo GM1 and sialic acid were without effect. In the standard test cultures, DNA synthesis declined very steeply after the first day, with cell numbers stabilizing at the level reached after 2 days. Ganglioside was not itself responsible for the restricted proliferative response, as serum produced the same behaviors.     

Simultaneous determination of specific antigen, antibody and immune complexes in the serum of patients with chronic schistosomiasis

The main steps of a sensitive and quantitative technique for the simultaneous determination, in the same microplate, of antigen, antibody and specific immune complexes has been described. The assay involved principles of the double sandwich technique used in the "Enzyme-Linked Immunosorbent Assay" (ELISA). The technique proved to be useful for the estimation of antigen, antibody and specific immune complexes in sera of patients with human chronic hepato-intestinal schistosomiasis.     

The complement system and circulating immune complexes in burn patients

The results of the study of humoral immunity in 80 burn patients are presented. A typical feature of all burn patients was a decrease in the total activity of the alternative path of the activation of complement and its factors B and D, beginning from the first day after the trauma. The character of changes in the functional activity of components C1-C5 of the classical path depended on the area of damages with their activation if burn area was less than 20% of the skin surface and deactivation if the burn area exceeded 20%.     

Activation of Rickettsiae prowazekii in the composition of immune complexes by antiglobulin serum

The antiglobulin serum (AGS) against spotted fever immunoglobulin of guinea pigs was studied for its effect on the in vitro created immune complex rickettsiae + antibody in experiments using the procedure of laboratory cultivation of Rickettsiae prowazekii according to Weigl. The biological activity of the pathogen is shown to be restored under the action of AGS. This fact may be used for detecting the antigen + antibody complexes in case of the spotted fever and for studying their role in the relapsive forms of the disease.     

Immunosuppression of T lymphocyte function by fractionated serum from tumor-bearing mice.

Sera from mice with transplanted 3-methylcholantrene-induced tumors have been shown previously to inhibit the function of normal lymphoid cells. When chromatographed on Sephadex G-150, the fraction eluting with immunoglobulin has been shown to inhibit the proliferative response of normal spleen cells to concanavalin A and to inhibit the in vitro antibody response to a T-dependent antigen, but has a lesser effect on the antibody response to a T-independent antigen. This paper deals with studies on the mode of action of the serum factor. The immunoglobulin containing fraction of serum from tumor-bearing mice inhibited the in vitro generation of both allogeneic and syngeneic cytotoxic lymphocytes. Time course studies demonstrate that the serum fraction inhibits the generation of antibody-producing and cytotoxic lymphocytes if added during the first 2 days of a 5-day culture. Serum fractions added after day 2 had no effect on the in vitro response. The serum factor appears to inhibit the generation of specific T cell function during the proliferative stage of development but has no effect on the differentiation stage which leads to either antibody-producing cells or cytotoxic lymphocytes.