Experimental arthropathy induced in rhesus monkeys and DBA/1 mice by a novel method: intraperitoneal implantation of type II collagen adsorbed onto nitrocellulose filters

A novel method which avoids the use of complete Freund's adjuvant (which can be arthritogenic) has been used to induce collagen II arthritis in both primates and mice. A solution of bovine type II collagen was dried onto nitrocellulose filters and implanted in the peritoneal cavity of experimental animals. Primate and mouse joints were scored by clinical as well as gross and microscopic parameters. The polyarthritis that developed in both rhesus monkeys (Macaca mulatta) and DBA/1 LAC J mice was characterized by synovial cell proliferation and endothelial cell hyperplasia, and by a perivascular mononuclear cell infiltrate of the synovium. Primates were analyzed further for anti-type II collagen antibody titers and delayed type hypersensitivity to type II collagen. Anti-type II collagen serum titers appeared to be unrelated to the disease pathology; the primates did not display delayed-type hypersensitivity to type II collagen. Control monkeys and mice implanted with collagen-free nitrocellulose filters were normal upon clinical and histopathological analysis. This protocol offers the advantage of the induction of arthritis due solely to immunization with antigen.     

Characterization of platelet-derived growth factor beta-receptor expressing cells in the vasculature of human rheumatoid synovium.

Platelet-derived growth factor (PDGF) beta-receptor expression in normal and rheumatoid synovia was investigated by double immunofluorescence staining of frozen sections and by in situ hybridization. In the inflamed synovia, PDGF beta-receptor mRNA was present in vascular cells, as well as in discrete stromal cells. PDGF beta-receptor expressing cells in rheumatoid synovia were characterized by double immunofluorescence staining using the PDGFR-B2 monoclonal antibody at a concentration at which this antibody merely stained granular accumulations of PDGF beta-receptors. Granular accumulations of PDGF beta-receptors were articulate in blood vessel cells, but also appeared in discrete stromal cells. Thus, the overall distribution of cells having granular accumulations of PDGF beta-receptors was similar to the distribution of cells expressing PDGF beta-receptor mRNA. Double immunofluorescence stainings showed that: (a) a majority (greater than 90%) of resident macrophages did not express granular PDGF beta-receptor staining, but macrophages were often juxtaposed to PDGF beta-receptor-positive cells; (b) T lymphocytes did not express PDGF beta-receptors, but these cells were frequently found in the proximity of cells stained by PDGFR-B2; (c) in some blood vessels both HLA-DR expressing cells and PDGF beta-receptor expressing cells could be visualized, whereas in other blood vessels, cells expressing only one of these activation markers could be detected; (d) smooth muscle cells in blood vessels contained PDGF beta-receptors; and (e) capillary endothelial cells in the inflamed synovia recurrently displayed granular PDGF beta-receptor staining. The granular accumulations of PDGF beta-receptors may reflect internalization of the receptor as a result of paracrine or autocrine ligand stimulation. In support of such a possibility are the findings that elevated levels of PDGF B chain mRNA were detected by in situ hybridization in the inflamed synovia, and that cells expressing PDGF B chain mRNA were distributed similarly to cells expressing PDGF beta-receptor mRNA. Taken together, the results indicate that PDGF has a role in the inflammatory process in rheumatoid synovitis, most likely by stimulating proliferative events in the vasculature.     

Pathogenesis of bloodstream invasion with Haemophilus influenzae type b

Possible route(s) by which encapsulated bacteria invade the blood from the nasopharynx include (i) the direct invasion of submucosal blood vessels and (ii) clearance via lymphatics to regional nodes followed by bloodstream invasion. These possibilities were investigated in rats after intranasal inoculation with 10(5) Haemophilus influenzae type b. Within 24 h of inoculation, 10 of 42 rats with sterile blood cultures had similar numbers of H. influenzae b recovered from both cervical (local) and periiliac (distant) lymph nodes, which suggested early bacteremic spread. When virtually continuous blood cultures were obtained for 30 min after inoculation with 10(8) H. influenzae b, early transient bacteremia was documented in four of eight rats. Also, we found no significant difference in bacteremia among rats whose cervical lymph nodes had been removed surgically compared with sham-operated rats. These findings favor the hypothesis of a rapid, perhaps direct invasion of pharyngeal blood vessels as an initial determinant of the systemic spread of H. influenzae b.     

Xenoserum-induced cytolytic "T" cells: polyclonal specificity with an apparent "anti-self" component, and cooperative induction.

Mice were primed in vivo by injection of fetal calf serum (FCS) and their spleen cells were incubated in vitro for 5 days in medium containing 10% FCS. This resulted in the development of cytolytic activity, which was most probably due to "T" cells, since effector cells 1) were sensitive to anti-Thy 1 antiserum or monoclonal antibodies in the presence of complement, 2) were not retained on Ig-anti Ig columns, 3) did not develop from "nude" spleen cells. Further arguments for the T cell nature of these effector cells came from their specificity. Blocking experiments using unlabeled competitor cells demonstrated that FCS-induced cytolysis was polyclonal, with clones recognizing allogeneic or syngeneic determinants possibly related to allo or self H-2. In keeping with polyclonality, cytolysis tested on any given target cell was greatly increased by adding Concanavalin A during the cytolysis test. Experiments were made to investigate whether in particular the anti-self cytolytic activity was directed against FCS determinants. We feel that this possibility, although not formally excluded, was made unlikely. The polyclonal specificity at the effector stage stood in sharp contrast to the serum specificity at the induction stage (reported elsewhere). We demonstrated that these two sets of specificities corresponded to two sets of specific cells. A first population of FCS-primed cells had "promoter" activity, in the sense that it could trigger a second population of "precursor" cells to differentiate into polyclonally cytolytic T cells.     

Lymphocyte proliferation in vitro induced by soluble protein antigens. II. Cellular requirements.

Immune guinea pig lymph node cells were fractionated on Ig anti-Ig or HSA anti-HSA affinity columns or on plastic surface in medium containing carbonyl iron. These techniques selectively removed B lymphocytes, K lymphocytes or adherent cells. The residual cells (Fc receptor-negative T lymphocytes) responded to soluble antigen in vitro in the same way or even better compared with nonfractionated cells. In addition, there was no indication that antigen-antibody complexes were superior to antigen in triggering lymph node cells or purified lymph code T lymphocytes into DNA synthesis. The results obtained suggested that memory T lymphocytes can be stimulated by antigen autonomously.     

Kappa Delta Award paper. Osteoregulatory nature of mechanical stimuli: function as a determinant for adaptive remodeling in bone

The capacity for functional adaptation within the skeleton was studied using the functionally isolated turkey ulna preparation. The results of this study would suggest that adaptive bone remodeling is extremely sensitive to alterations in both the magnitude and distribution of the strain generated within the bone tissue. At present, it appears that a loading regime can only influence bone remodeling when it is dynamic in nature. The full osteogenic potential of its influence is then achieved after only an extremely short exposure to this stimulus. The potency of the stimulus appears to be proportional to the magnitude of the strain engendered. As strain levels that are acceptable in one location induce adaptive remodeling in others, it would appear that each region of each bone is "genetically programmed" to accept a particular amount and pattern of intermittent strain as "normal." Deviation from this "optimal strain environment" will stimulate changes in the bone's remodeling balance, resulting in adaptive increases or decreases in its mass.     

The Mexican asthma cure. Systemic steroids for gullible gringos

Asthmatic patients from western Canada and the United States have reported that after visits to an asthma clinic in Mexicali, Mexico, they return home substantially improved or cured having received "a bronchodilator medication unavailable in the United States or Canada because of the big drug companies." Analysis of these medications reveals that the most commonly prescribed combination is the glucocorticoid triamcinolone (unscored white tablets) and the antihistamine chlorpheniramine (coated biconvex orange or red tablets). Occasionally benzodiazepines are added to these medications. The patients are assured that the medications which they have been given are free of side effects and specifically, that corticosteroids are not used. Such therapy is dangerous to the patient who not only is unaware of the medications that he or she is taking, but is unlikely to mention this therapy to his or her physician. These patients risk drug interactions, medication side effects, and the possibility of adrenal failure either with a stress to their system or on withdrawal of drug treatment. Patients are also at risk of abandoning safer forms of asthma therapy for the miracle cure. We, too, are partially responsible for these unethical practices by avoiding the use of steroids and undertreating our patients at times, leaving them unnecessarily restricted and eager for any form of relief.     

Analysis of cis-acting requirements of the Rh3 and Rh4 genes reveals a bipartite organization to rhodopsin promoters in Drosophila melanogaster

The rhodopsin genes of Drosophila melanogaster are expressed in nonoverlapping subsets of photoreceptor cells within the insect visual system. Two of these genes, Rh3 and Rh4, are known to display complementary expression patterns in the UV-sensitive R7 photoreceptor cell population of the compound eye. In addition, we find that Rh3 is expressed in a small group of paired R7 and R8 photoreceptor cells at the dorsal eye margin that are apparently specialized for the detection of polarized light. In this paper we present a detailed characterization of the cis-acting requirements of both Rh3 and Rh4. Promoter deletion series demonstrate that small regulatory regions (less than 300 bp) of both R7 opsin genes contain DNA sequences sufficient to generate their respective expression patterns. Individual cis-acting elements were further identified by oligonucleotide-directed mutagenesis guided by interspecific sequence comparisons. Our results suggest that the Drosophila rhodopsin genes share a simple bipartite promoter structure, whereby the proximal region constitutes a functionally equivalent promoter "core" and the distal region determines cell-type specificity. The expression patterns of several hybrid rhodopsin promoters, in which all or part of the putative core regions have been replaced with the analogous regions of different rhodopsin promoters, provide additional evidence in support of this model.