Simple Method for Enhancing Development of Acute Disseminated Encephalomyelitis in Mice.

Abstract

MURINE ACUTE DISSEMINATED ENCEPHALOMYELITIS 263 0.26. Metachromasia was tested with toluidine blue. None of the supernatant fluids from synovial tissue cultures and control developed metachromasia. Heparin (as little as 5 pg per ml) and other sulfonated mucopolysaccharides showed strong metachromasia with toluidine blue.Electrophoretic Studies. To further establish the nature of the mucopolysaccharides found in the supernatants, electrophoretic patterns were obtained with a Perkin-Elmer Tiselius apparatus.Supernates with negative acetic acid precipitation tests gave protein patterns with peaks characteristic of the globulins and albumin. The latter gave the highest and fastest moving peak (Fig. la). Supernates which gave a positive precipitation test showed a new peak in front of the albumin (Fig. lb). The mobility of this component was in the range of hyaluronic acid verified by the addition of purified hyaluronic acid to a negative supernatant fluid (Fig. lc). None of the supernates exhibited a peak with a faster mobility than that for hyaluronic acid. The sulfonated mucopolysaccharides such as chondroitin sulfuric acid or heparin have a faster mobility than the hyaluronic acid. This is seen in the electrophoretic pattern obtained when heparin is added to a tissue culture supernate positive for hyaluronic acid (Fig. Id).1. Growth of human and animal synovial membrane in tissue cultures showed characteristic differences from control cultures of periarticular tissues. 2. Presence of hyaluronic acid in about two-thirds of supernates of tissue cultures. from synovium was demonstrated by precipitation reactions with acetic acid, enzymatic reactions, viscosity determination, tests for metachromasia and electrophoretic patterns. 3. Quantitative determinations by a turbidimetric method yielded a concentration of hyaiuronic acid from 4 to 15 mg %. 4. Tests were negative for sulfonated mucopolysaccharides such as chondroitin sulfuric acid or heparin. 5. Control cultures of periarticular connective tissue were negative for hyaluronic acid and sulfonated mucopolysaccharides.

Pertussis Toxin Modulates the Immune Response to Neuroantigens Injected in Incomplete Freund’s Adjuvant: Induction of Th1 Cells and Experimental Autoimmune Encephalomyelitis in the Presence of High Frequencies of Th2 Cells

“…This effect of the toxin has been interpreted in the past as a result of its action on the BBB. The model held that PT opened up the BBB by histamine-induced vascular leakage (17) and thereby facilitated the transmigration of T cells and the induction of disease (7,37). However, this interpretation has been challenged (18,38), and our data support an additional mechanism of its action as well.…”

“…In contrast, injection of the same myelin Ags in IFA is highly efficient in protecting from EAE (5,6). Interestingly, the incidence and severity of the disease induced by immunization with neuroantigens in CFA is enhanced by the coinjection of pertussis toxin (PT) (7)(8)(9).…”

IFN-γ-Induced Chemokines Synergize with Pertussis Toxin to Promote T Cell Entry to the Central Nervous System

“…Experimental autoimmune encephalomyelitis (EAE) is an animal model for MS. Infectious agents or their products are frequently used as adjuvants in EAE induction protocols. The toxin produced by Bordetella pertussis toxin (Ptx) is one such agent, used in EAE induction since 1955 (4). The generally held view has been that Ptx acts to "open up the bloodbrain barrier," based on evidence that Ptx increases vascular permeability, in part by enhancing vascular sensitivity to vasoactive amines (5,6).…”

Effect of Nutrition on the Production of Acute Disseminated Encephalomyelitis in Mice

“…Distilled water was supplied ad libitum. Production of ADE in Mice.--The empirical basis of the production of ADE in mice has been subject to a continuing process of revision in recent years (15,11,12). During the investigations reported here one such revision was adopted so that, in course, two techniques were employed.…”

“…Historically, the development of this particular phenomenology has proceeded from a recognition of the consequences of injection into hosts of homologous and heterologous brain material (1, 2), the properties of adjuvants in enhancing the pathological effects of such injections (3,4), the more precise chemical definition of the injected brain material incitant (5)(6)(7)(8), the efficacy of various modes of injection (9), the role of hypersensitivity (10,11), with enhancement by Hemophilus pertussis vaccine (12), and the recognition of the genetic control of host susceptibility with a consequent Mendelian analysis of resistant and susceptible mouse genotypes (11,13). During the course of the genetic analysis evidence appeared which suggested that still another facet of this expanding phenomenology was open to experimental attack; namely, the effect of host nutrition on the capacity to elicit the disorder.…”