Monoclonal immunoglobulins in patients with carcinoma of the colon

Summary Structural studies were performed on five monoclonal immunoglobulins isolated from patients with carcinoma of the colon. Serologic analysis revealed that two of the five proteins shared idiotypic antigenic determinants; these two but none of the three others had V_HIII heavy chains. The results demonstrated a close structural similarity between the heavy chains of these two proteins both in their antigen-binding sites (the hypervariable region) and in their framework regions. Determination of the NH₂-terminal amino acid sequences indicated that the light chain variable regions of all five proteins were either V_xII or V_xIII. These data suggest that monoclonal immunoglobulins in patients with carcinoma of the colon have restricted heterogeneity and that, in some cases the production of monoclonal immunoglobulin(s) and the development of a solid tumor in a given patient may be related events.Publication no. 357 from the Department of Basic and Clinical Immunology and Microbiology, Medical University of South Carolina. Research supported in part by National Science Foundation Research Grant PCM 79-24043, USPHS Grants HD-09938 and CA-25746, and by Medical University of South Carolina Biomedical Research Appropriations A911 and A912. A. C. W. was the recipient of American Cancer Society Faculty Research Award No. 125     

酪氨酸激酶Btk在根尖周炎中的表达研究

目的 研究大鼠根尖周炎过程中Btk的表达,分析其与患牙根尖区骨吸收的关系,探讨根尖周炎患牙骨吸收的机制。方法 体内部分收集人慢性根尖周炎病损组织15例,体外构建大鼠慢性根尖周炎模型20例,采用实时荧光定量PCR检测Btk mRNA的表达情况。结果 Btk在人慢性根尖周炎病损组织中表达较根尖周正常组织明显增高。实验组大鼠根尖区Btk的表达呈倒V型,即在前2周呈上升趋势,并于2周时达到顶峰,然后开始逐渐降低。结论 Btk参与了根尖周炎的发生和发展过程。     

Prophylaxis and therapy with human immunoglobulins

The supply of human immunoglobulin preparations (Ig) is steadily increasing. On the one hand, the wide range of specificities of the so-called hyper-Ig is broadening, on the other hand, the structural variety of Ig contained in the preparations and their fragments is increasing (intact IgG, Fab, F(ab')2 fragments, desaggregation, inactivation of the complement binding site). Specificity and structure of Ig are important for the efficacy, kind of application, amount and intervals of dosage and for compatibility. Therefore, an exact knowledge of the preparation becomes more and more necessary for reasonably applying it. The development of modern IgG preparations with intravenous compatibility and intact Fc-part providing the possibility for the application of a high dosage require "old" indications to be re-examined by clinically controlled investigations, thus opening new ways to an improved efficacy in applying Ig in the prophylaxis and therapy of infections as well as to new areas of indication by recognising the immunomodulatory effects, e.g. autoimmunopathies.     

Monoclonal antibody to serum immunoglobulins of Clarias batrachus and its application in immunoassays

Serum immunoglobulins of Clarias batrachus (Cb-Ig) were purified by affinity chromatography using bovine serum albumin as capture ligand. Under reducing conditions in SDS-PAGE, Cb-Ig was composed of a heavy (H) chain (68.7 kDa) and two light (L) chains (27.4 and 26.3 kDa). Purified Cb-Ig was used to produce a monoclonal antibody (MAb) designated E4 MAb that belonged to IgG1 subclass. In Western blotting, this MAb showed binding to H chain of purified Cb-Ig and putative H chains in reduced sera of C. batrachus, Clarias gariepinus and Heteropneustes fossilis. However, no binding was observed with serum protein of Labeo rohita and Channa striata. Cross-reactivity of anti-Cb-Ig MAb was observed with serum of C. batrachus, C. gariepinus and H. fossilis in competitive ELISA. In immunoblotting of non-reduced Cb-Ig with E4 MAb, four bands assumed to be tetrameric, trimeric, dimeric and monomeric form were observed. In flow cytometric analysis of the gated lymphocytes, the number of surface Ig-positive (Ig +) cells in blood, spleen, kidney and thymus of C. batrachus was determined to be 50.1 ± 3.1, 55.1 ± 3.36, 42.4 ± 4.81 and 5.1 ± 0.89%, respectively, using E4 MAb. Ig + cells were also demonstrated in formalin-fixed paraffin embedded tissue sections of spleen, kidney, thymus and smears of blood mononuclear cells in indirect immunoperoxidase test. The developed MAb was employed to detect pathogen-specific immunoglobulins in the sera of C. batrachus immunized with killed Edwardsiella tarda, by an indirect ELISA. This monoclonal antibody can be useful tool in immunological research and assays.     

Study of the antigenic relationship between animal and human immunoglobulins using antisera to human immunoglobulins]

Cross-reactivity of monospecific antisera to human immunoglobulins with animal sera of 10 species was studied by immunoelectrophoresis and radial immunodiffusion. Antisera to IgG were shown to reveal IgG of all the species studied, antisera to IgM and especially to IgA cross reacted less extensively. The greatest number of cross reactions were given by the antisera obtained as a result of hyperimmunization. Hyperimmune monospecific antisera to human IgG, IgA, and IgM can be used for the identification of animal immunoglobulins during their isolation from the sera and for their quantitation by radial immunodiffusion.     

Characterization of a localized basophil hypersensitivity lesion in guinea pig conjunctiva

Basophils accumulate in response to antigen challenge in cutaneous basophil hypersensitivity (CBH) reactions. Two ocular diseases, vernal conjunctivitis and contact-lens-associated conjunctivitis, are also characterized by this histopathology. We have refined a model previously developed in guinea pig conjunctiva by precisely defining the site of antigen injection and correlating the site with the clinical and histologic changes. Guinea pigs were primed by an intradermal injection of keyhole limpet hemocyanin (KLH) in the flank and challenged (Day 6) by injection of a small bolus of KLH just under the conjunctival epithelium. Twenty-four hours later histologic examination showed a perivascular infiltrate of inflammatory cells containing large numbers of basophils. Eosinophils, neutrophils, macrophages, and mast cells were also seen. Serial sections of the reaction site showed discrete boundaries. At all sites examined in antigen-challenged tissues, there were significantly more basophils than in control-injected conjunctiva. Insertion of a sterile needle or injection of PBS or KLH into normal conjunctiva induced a significant increase in neutrophils and some macrophages. Injection of graded doses of antigen into the conjunctiva of primed animals, resulted in a dose-dependent increase in basophils up to 50 micrograms KLH (optimal dose).     

Co-purification of soluble human galactosyltransferase and immunoglobulins

Preparations of human malignant effusion galactosyltransferase activity purified according to previously published techniques using enzyme-specific affinity chromatography consistently produced antibodies directed toward immunoglobulins with no detectable antigalactosyltransferase. Double immunodiffusion analysis of the antigen showed the presence of both IgG and IgA. Affinity chromatography with anti-human IgG-Sepharose and anti-human serum-Sepharose resulted in a 48,000-fold purification of galactosyltransferase activity with no detectable IgG by radioimmunoassay. Immunization of rabbits with this preparation produced antibodies directed against galactosyltransferase activity and minimal anti-Ig. The persistence of immunoglobulins during the purification of soluble galactosyltransferase activity through two enzyme-specific affinity chromatographic steps suggests an association of immunoglobulins with galactosyltransferase activity.     

Monoclonal antibodies to bovine and human acrosin

Monoclonal antibodies to human acrosin were required for studies of immunological interference with fertilization. Since human acrosin was not available in adequate amounts, monoclonal antibodies have been raised in mice against purified bovine acrosin and screened for cross-reaction with human sperm cells. Two of these antibodies are described, B4F6 and C2E5. Data from enzyme-linked immunosorbent assays, immunoblots, immunoprecipitation, and indirect immunofluorescence on sperm cells indicate that B4F6 binds only to bovine acrosin, and that C2E5 binds both to bovine and to human acrosin at a conformationally determined epitope. The antibodies do not inhibit the hydrolysis of benzoylarginine ethyl ester by acrosin, but C2E5 did inhibit the dissolution of the hamster zona pellucida by purified human acrosin. The antibodies have also been used for affinity purification of acrosin and proacrosin.     

Monoclonal antibodies and therapy of human cancers

This survey is an overview of the applications of murine, humanized and recombinant monoclonal antibodies for in vivo diagnostic and therapeutic applications. Monoclonal antibodies (mAb) have been applied to the diagnosis and therapy of an array of human diseases. The initial failures of early clinical trials have been overcome through the production of a new generation of mAb which features reduced immunogenicity and improved targeting abilities. The early models of mAb therapy were focused on enhancing the cytolytic mechanisms against the tumor cells. More recently, successful mAb-based therapies were targeted to molecules involved in the regulation of growth of cancer cells. This has highlighted the relevance of understanding receptor-mediated signaling events, and may provide new opportunities for anti-tumor antibody targeting. Despite all the difficulties, clinical data is outlining an increasingly significant role for antibody-mediated cancer therapy as a versatile and powerful instrument in cancer treatment. One reasonable expectation is that treatment at an earlier stage in the disease process or in minimal residual disease may be more advantageous.