Off label pharmacological therapy in patients with short bowel syndrome.

BACKGROUND: Short bowel syndrome is a disabling disease requiring long-term nutritional support and ancillary drugs. Aiming to analyze the most commonly prescribed drugs, a retrospective analysis was conducted is an outpatient cohort. PATIENTS AND METHODS: Stable patients (N= 37, 59.5% males, age 51.1 ± 20.1 years, body mass index 20.1 ± 7.9 kg/m2) with three or more appointments in the Outpatient Service during the last 18 months were retrospectively analyzed. regarding oral pharmacologic prescriptions. Medications were classified as on label or off label. RESULTS: A total of 257 oral prescriptions were retrieved from computer files, encompassing 17 different preparations. The majority was employed on label however 28.8% (74/257) were classified as off label and scrutinized with regard to indications. The main categories were pharmacologic modulators of gastrointestinal secretions and motility, along with antibiotics. Virtually all patients required one or more of such drugs, without differences regarding demographic or clinical variables. Adverse effects or premature drug discontinuation were not observed. CONCLUSIONS: This is the first study to our knowledge highlighting the importance of adjuvant drugs, particularly with unconventional indications, in the management of short bowel syndrome. Antidiarrheic agents, pancrelipase micropellets, antacids and antibiotics represented the most relevant off label prescriptions for this population.

A phase 1 study of a meningococcal native outer membrane vesicle vaccine made from a group B strain with deleted lpxL1 and synX, over-expressed factor H binding protein, two PorAs and stabilized OpcA expression.

This phase I clinical trial assessed the safety and immunogenicity of a native outer membrane vesicle (NOMV) vaccine prepared from an lpxL1(-) synX(-) mutant of strain 8570(B:4:P1.19,15:L8-5) of Neisseria meningitidis. Additional mutations enhance the expression of factor H binding protein variant 1 (fHbp v.1), stabilize expression of OpcA and introduce a second PorA (P1.22,14). Thirty-six volunteers were assigned to one of four dose groups (10, 25, 50 and 75 mcg, based on protein content) to receive three intramuscular injections at six week intervals with aluminum hydroxide adjuvant. Specific local and systemic adverse events were solicited by diary and at visits on days 2, 7, and 14 after each vaccination. Blood chemistries, complete blood count, and coagulation studies were measured on each vaccination day and again 2 and 14 days later. Blood for ELISA and serum bactericidal assays was drawn two and six weeks after each vaccination. The proportion of volunteers who developed a fourfold or greater increase in bactericidal activity to the wild type parent of the vaccine strain at two weeks after the third dose was 27 out of 34 (0.79, 95% C.I. 0.65-0.93). Against four other group B strains the response rate ranged from 41% to 82% indicating a good cross reactive antibody response. Depletion assays show contributions to bactericidal activity from antibodies to lipooligosaccharide (LOS), fHbp v.1 and OpcA.

Restricting expression prolongs expression of foreign genes introduced into animals by retroviruses.

If foreign genes are ubiquitously expressed in mice using a viral vector, expression is abrogated by CD8(+) cells in 2 to 4 weeks. However, if the expression of the genes is confined to skeletal muscle cells, the CD8(+) T-cell response is much weaker and expression is maintained for more than 6 weeks. These data show that restricting the expression of foreign genes to skeletal muscle cells and presumably to other cells that are inefficient at antigen presentation can prolong the expression of a foreign gene product.

Extracellular matrix-induced stimulation of a CD4-8- thymic T cell line.

PBK101 was a thymic T cell line of AKR/J origin that expressed Thy 1, CD44, IL-2R, and VLA-4 antigens but not CD4, CD8, or the TCR/CD3 complex. PBK101 was initially isolated growing together with adherent thymic stromal cells. The proliferation of PBK101 ceased when it was cultured in the absence of stromal cells and its proliferation was stimulated when stromal cells were added back to the culture. PBK101 cells were also stimulated when cocultured with peritoneal exudate cells (PEC). These responses appeared to require contact between PBK101 and the cellular stimulators. The stimulatory activity of the thymic stromal cell line remained in culture wells after the cellular elements were removed by mild trypsinization or detergent extraction indicating that a stimulatory molecule(s) was associated with extracellular matrix (ECM). The activity of the ECM was resistant to high salt extraction, acetic acid, and strong denaturants but was sensitive to treatment with trypsin, heat (80 degrees C), and, if cells were treated with a lathyragen, 4 M guanidine-HCl plus 2 mM dithiothreitol (DTT). The extracellular matrix produced by some other fibroblast cell lines was nonstimulatory. Stimulation of PBK101 by ECM was not inhibited by monoclonal anti-VLA4 or polyclonal anti-fibronectin antisera. Purified extracellular matrix proteins (laminin, collagen, fibronectin, and vitronectin) were either nonstimulatory or only marginally stimulated PBK101 cells. These results suggested that a potentially novel molecule(s) present in the extracellular matrix of a thymic stromal cell line and on the surface of other cellular elements stimulated a thymic T cell line of immature phenotype. The potential role of extracellular matrix-associated molecules in normal T cell development is discussed.

Anti-Ig inactivation of the CH31 lymphoma model for immature B cell inhibits its ability to process pigeon cytochrome c.

We wished to determine whether tolerized cells are able to process and present antigen based on our hypothesis that tolerized B cells be unable to function in the normal capacity as antigen-presenting cells if they are to remain the tolerant state. Results in this study show that the ability of the murine lymphoma model for immature B cells, CH31, to process pigeon cytochrome c was greatly down-regulated when cultured in the presence of rabbit anti-mouse IgM. In contrast, the same anti-IgM treatment had no significant effect on the antigen-presenting cell function of the lymphoma model for mature B cells, CH112. Presentation of CNBr-cleaved fragments of pigeon cytochrome c by either CH31 or CH12 cells was not affected by the antibody treatment. Furthermore, CH31 cells pre-incubated with pigeon cytochrome c were not subject to the anti-IgM inhibition of the antigen presentation. These observations suggest that pertubation of surface immunoglobulin molecules on CH31 immature B cells causes down-regulation of their antigen-processing machinery.

Characterization of the proliferative response of a CD4-8- thymic T lymphoma cell line to stimulation by thymic cellular elements.

E710.2 is a cloned T cell line that was isolated from an AKR/J thymic tumor. This clone expresses Thy-1, heat-stable Ag, and the CD3/TCR complex but does not express CD4 or CD8. When the E710.2 cell line is injected into syngeneic mice, it grows as a malignant tumor in lymphoid organs and the thymus. In contrast, this cell line does not grow in vitro under standard culture conditions. This latter property allowed us to analyze the in vitro responsiveness of this CD4-CD8- cell line to stimulation by pharmacologic agents and cellular elements from the spleen and thymus. E710.2 cells proliferate when stimulated with phorbol esters or when cocultured with thymocytes or splenocytes. We could not detect soluble stimulatory factors in cultures of E710.2 and/or lymphoid cells, suggesting that cell contact might be required for this response. The stimulatory activity in thymus and spleen appears to be broadly expressed, because all cell subsets that were examined from these tissues stimulate this cell line. The stimulation of E710.2 cells is not MHC-restricted and is not inhibited by anti-MHC mAb. Furthermore, the responsiveness of these cells is not decreased when the TCR/CD3 complex is modulated from the cell surface. Similarly, TCR/CD3-deficient E710.2 variant clones retain their responsiveness to thymic and splenic cell stimulation. These findings suggest that there is a TCR-independent pathway of activation in E710.2 that is stimulated by a broadly expressed, non-MHC-encoded molecules(s).

Lectins and antibodies as tools for studying cellular interactions.

Specific interactions between multiple cell types are critical for a variety of processes central to the development, homeostasis and immune defense of multicellular organisms. Studies designed to elucidate how cells communicate through physical encounters have exploited exogenously supplied factors to bypass intrinsic recognition mechanisms and facilitate cellular conjugation. In this review, we compare the relatively nonspecific agglutinating properties of lectins and the selective cell targeting capabilities of antibodies and bispecific antibody constructs for studying cell-cell interactions in immunobiology. In addition, we discuss a novel system for inducing cellular interactions which closely resembles native receptor-mediated conjugation. In this system, surrogate receptors promote specific cell-cell interactions without hindering endogenous receptor-ligand interactions at the cell-cell interface which may be important in mediating physiologic cellular responses.

Effects of interleukin 4 on neonatal B lymphocyte tolerance.

Perturbation of antigen receptors on mouse neonatal B cells by rabbit antimouse IgM antibody was shown to inhibit cell proliferation in response to the B cell mitogen lipopolysaccharide. When these antibody-inactivated cells were challenged with lipopolysaccharide in the presence of the helper T cell product interleukin 4, a strong proliferative response was observed. Interleukin 4 alone did not cause proliferation of the antibody-treated B cells. Pretreatment with interleukin 4 did not prevent neonatal B cell inactivation by the antibody. Our results show that neonatal B cells inactivated directly through their antigen receptors can be reactivated by the combined signals of interleukin 4 and lipopolysaccharide.

Purification, partial characterization, and clinical evaluation of an adenocarcinoma-associated antigen.

The current investigation describes the purification and partial characterization of a new adenocarcinoma-associated antigen (ACAA). ACAA is a large molecular weight glycoprotein (Mr 790,000 by size chromatography on Sepharose CL-6B) that migrates in the alpha 1 region upon electrophoresis and is eluted from a DEAE-cellulose column at a 0.1 M NaCl concentration. ACAA is immunochemically and biochemically different from carcinoembryonic antigen, alpha-fetoprotein, pancreatic oncofetal antigen, human pancreatic tissue antigen, CA 19-9, ferritin, and acute-phase proteins. Assays for ACAA were carried out using a solid-phase sandwich enzyme immunoassay. The results indicate that ACAA is present in sera of all individuals. Patients with cancer have higher serum levels of ACAA than normal individuals. The greatest frequency of elevated serum values of ACAA was seen in patients with lung and pancreatic cancers followed by colorectal, breast, and prostate cancer. The measurement of ACAA levels may be valuable in the diagnosis and clinical management of patients with certain cancers.