4i
`
`A_ (cid:9)
`
`• (cid:9)
`
`'
`
`CLINICAL AND EXPERIMENTAL IMMUNOLOGY
`
`VOLUME 7 . NUMBER 1 . JULY 1970
`
`CONTENTS
`
`F. D. LINDSTROm: Kappa: lambda light chain ratio in IgG eluted from rheumatoid
`arthritis synovium
`
`J. A. AARLI & 0. ToNDER: Antiglobulin consumption tes with sera from patients with
`myasthenia gravis'
`
`J. A. AARLI: Binding of y-globulin fragments to muscle tissue
`
`MARGARET TURNER WARWICK & PATRICIA HASLAM: Smooth muscle antibody in
`bronchial asthma
`
`M. INouE & G. KLEIN: Reactivity of radioiodinated serum antibody from Burkitt's
`lymphoma and nasopharyngeal carcinoma patients against culture lines derived
`from Burkitt's lymphoma
`
`I. A. MCGREGOR, D. S. RoWE, M. E. WILSON & W. Z. BILLEWICZ: Plasma immuno-
`globulin concentrations in an African (Gambian) community in relation to season,
`malaria and other infections and pregnancy (cid:9)
`
`1
`
`11
`
`23
`
`31
`
`35
`
`51
`
`B. M. GREENWOOD, ERINA M. HERRICK & E. J. HOLBOROW Speckled antinuclear
`factor in African sera
`g.
`
`75 -
`
`G. VIRELLA & MARIA F. LOPES-VIRELLA: Effects of therapeutically useful thiols (DL-
`penicillamine and oc-mercaptopropionylglycine) on immunoglobulins
`
`85
`
`H. W. KRETH, G. THIESSEN & H. DEICHER: The effect of antigen given intravenously
`on specific antigen-sensitive lymphocytes of peripheral blood in man (cid:9)
`
`109
`
`A. PASTERNACK & E. LINDER: Renal tubular acidosis : an immunopathological study on
`four patients (cid:9)
`
`115
`
`
`CASE
`
`REPORT
`
`M. I. DRURY, DEBORAH M. KEELAN, F. J. TIMONEY & W. J. IRVINE: Juvenile familial
`endocrinopathy (cid:9)
`
`125
`
`BLACKWELL SCIENTIFIC PUBLICATIONS
`OXFORD AND EDINBURGH
`
`Printed and bound in Great Britain by Alden & Mowbray Ltd
`at the Alden Press, Oxford (cid:9)
`•
`
`N a
`
`',S.
`
`CLINICAL AND
`EXPERIMENTAL
`IMMUNOLOGY
`
`EDITOR W. J. IRVINE
`
`Blackwell Scientific Publications
`Annual Subscription £18 ($60.00) post free
`Single issues at 35s ($6.00) net
`VOL 7 NO 2 AUGUST 19\70
`An official journal of the
`British Society for Immunology
`
`Ex. 1045 - Page 1
`
`

`

`CLINICAL AND EXPERIMENTAL IMMUNOLOGY
`EDITOR: W. J. IRVINE
`
`EDITORIAL BOARD
`J. F. ACICROYD, St Mary's Hospital, London, W.2, England
`G. L. ASHERSON, Department of Bacteriology, The London Hospital, London, E.1, England
`H. BENNICH, The Wallenberg Laboratory, Box 562, S-751 22 Uppsala 1, Sweden
`C. G. COCHRANE, Scripps Clinic and Research Foundation, La Jolla, California, U.S.A.
`R. R. A. COOMBS, Department of Pathology, University of Cambridge, England
`J. DAussrr, Institut de Recherches de la Faculte de Medecine, Hopital Saint-Louis, Paris Xe, France
`A. M. DENMAN, Division of Immunology, M.R.C. Clinical Research Centre, Northwick Park, Middlesex,
`England
`DEBORAH DONIACH, The Middlesex Hospital, London, England
`H. H. FUDENBERG, Department of Medicine, San Francisco Medical Center, San Francisco, California, U.S.A.
`J. L. GOWANS, Cellular Immunology Research Unit, Oxford, England
`R. GRUBB, Bakteriologiska Institutionen, Lund, Sweden
`R. J. C. HARRIS, Department of Environmental Carcinogenesis, Imperial Cancer Research Fund, London,
`England
`K. HIRSCHHORN, Division of Medical Genetics, The Mount Sinai School of Medicine, 100th Street and Fifth
`Avenue, New York, U.S.A.
`J. G. HOWARD, Department of Experimental Immunobiology, Wellcome Research Laboratories, Langley
`Court, Beckenham, Kent, England
`K. JAMES, Department of Surgical Science, Edinburgh University Medical School, Scotland
`M. H. KAPLAN, Department of Medicine, Cleveland Metropolitan General Hospital, Cleveland, Ohio, U.S.A.
`G. KLEIN, Department of Tumour Biology, Karolinska Institutet, Stockholm, Sweden
`I. R. MACKAY, The Walter & Eliza Hall Institute of Medical Research, Melbourne, Australia
`P. A. MIESCHER, Division of Haematology, Department of Medicine, Hopital Cantonal, 25 rue Michell-du-
`Crest, Geneva, Switzerland
`J. F. A. P. MILLER, Department of Experimental Pathology, The Walter & Eliza Hall Institute of Medical
`Research, Melbourne, Australia
`R. C. NAIRN, Department of Pathology, Monash University, Alfred Hospital, Prahran, Victoria, Australia
`P. PERLMANN, The Wenner-Gren Institute, Stockholm, Sweden
`J. H. RoCKEY, Department of Ophthalmology, School of Medicine, University of Pennsylvania, Philadelphia
`19104, U.S.A.
`N. R. ROSE, Department of Bacteriology and Immunology, State University at Buffalo, New York, U.S.A.
`R. S. SCHWARTZ, Clinical Immunology Service, New England Medical Center Hospitals, 171 Harrison
`Avenue, Boston, Massachusetts 02111, U.S.A.
`M. SELIGMANN, Institut de Recherches sur les Maladies de Sang, Hopital Saint-Louis, Paris X°, France
`J. F. Sooran.L, Department of Immunology, London University Institute of Child Health, Great Ormond
`Street, London, England
`T. E. STARZL, Department of Surgery, University of Colorado Medical Center, Denver, U.S.A.
`M. W. TURNER, Department of Immunology, Institute of Child Health, 30 Guilford Street, London, W.C.1,
`_ •
`England
`J. J. VAN LOGHEM, Centraal Laboratorium van de Bloedtransfusiedienst, Amsterdam, The Netherlands
`J. WALDENSTRoM, Allmanna Sjukhuset, Malmo, Sweden
`D. M. WEIR, Immunology Unit, Department of Bacteriology, University of Edinburgh, Scotland
`SIR MICHAEL WOODRUFF, Department of Surgical Science, University of Edinburgh, Scotland
`
`EDITORIAL SECRETARY: Miss Patricia Burnett, B.Sc.
`Clinical and Experimental Immunology is published monthly, each issue consisting of about 150 pages. Six issues
`form one volume. The annual subscription is £18 (U.S.A. and Canada $60.00) post free; price per number 35s
`(U.S.A. and Canada $6). The journal is available to members of the British Society for Immunology at a special
`rate through the Treasurer of the Society.
`Editorial correspondence should be directed to the Editor, Dr W. J. Irvine, Department of Endocrinology, The
`Royal Infirmary, Edinburgh EH3 9YW, Scotland.
`Business matters, including correspondence and remittances relating to subscriptions, back numoers and
`offprints, should be sent to the publishers: Blackwell Scientific Publications Ltd, 5 Alfred Street, Oxford OX1
`4HB.
`Advertisement bookings and enquiries to S. & H. Fretwell Ltd, 267 Portland Road, London, S.E.25.
`Offprints. Fifty are supplied free and additional cop es may be ordered on the printed card which will be sent
`to the author with the proofs.
`
`Clinical and Experimental Immunology
`
`Clinical and Experimental Immunology
`
`Papers to be published in Vol. 7, No. 3, September 1970
`
`A. D. M. BRYCESON, R. S. BRAY, R. A. WOLSTENCROFT & D. C. DUMONDE:
`Immunity in cutaneous leishmaniasis of the guinea-pig
`
`A. J. AMMANN, H. J. MEUWISSEN, R. A. GOOD & R. HONG: Successful
`bone marrow transplantation in a patient with humoral and cellular
`immunity deficiency _
`
`N. R. ROSE & H. S. BONSTEIN: Trachea-specific antigens in normal and
`malignant human tissues
`
`0. ZETTERVALL & H. LINK: Electrophoretic distribution of kappa and
`lambda immunoglobulin light chain determinants in serum and cere-
`brospinal fluid in multiple sclerosis
`
`BRITTA WARREN & D. METCALF: Cytotoxicity in vitro of preleukaemic
`lymphoid cells on syngeneic monolayers of embryo or thymus cells
`
`DELPHINE M. V. PARROTT, MARIA A. B. DE SOUSA, VALERIE WALLIS,
`J. FACHET, ELIZABETH LEUCHARS & A. J. S. DAVIES: The response
`of normal thymectomized and reconstituted mice in contact sensitivity
`
`E A. CASPARY, D. HUGHES & E. J. FIELD: On the mode of action of anti-
`lymphocytic serum: experiments on electrophoretic mobility of macro-
`phages in experimental allegic encephalomyelitis
`
`D. L. BROWN, P. J. LACHMANN & J. V. DACIE: The in vivo behaviour of
`complement-coated red cells : studies in C6-deficient, C3-depleted and
`normal rabbits
`
`ANNA-BRITA LAURELL, ANDERS SJoHOLM & ULF JOHNSON: Quantitation
`of the fourth complement component by electrophoresis in agarose
`gel containing antibodies
`
`U. JUNGE, J. HOEKSTRA, L. WOLFE & F. DEINHARDT: Microtechnique for
`quantitative evaluation of in vitro lymphocyte transformation
`
`Ex. 1045 - Page 2
`
`(cid:9)
`

`

`Clin. exp. Immunol. (1970) 7, 139-145.
`
`ADJUVANT-ACTIVITY OF `DIPHTHEROID'
`ORGANISMS ISOLATED FROM THE JOINTS
`OF CASES OF RHEUMATOID ARTHRITIS
`
`R. G. WHITE AND J. GORDON
`
`Department of Bacteriology and Immunology,
`University of Glasgow
`
`(Received 9 February 1970)
`
`SUMMARY
`
`Two isolates of `diphtheroid' organisms from the joints of cases of rheumatoid
`arthritis were found to possess a surface network of filaments (125 A wide)
`resembling the adjuvant-active peptidoglycolipid filaments of mycobacteria and
`some Nocardia spp. Tests for adjuvant activity in guinea-pigs showed that both
`isolates possessed the ability to induce delayed-type hypersensitivity to a simul-
`taneously injected immunogen (ovalbumin) and to increase serum anti-ovalbumin
`levels (in particular y2-immunoglobulin). The relationship of adjuvant-active
`bacilli to the pathogenesis of rheumatoid arthritis is discussed.
`
`INTRODUCTION
`
`Recent isolations of mycoplasma from human cases of rheumatoid arthritis has re-awakened
`interest in the possible role of infection in this disease (Arai, Ishikawa & Hotta, 1964;
`Bartholomew, 1965). A later report by Duthie et al. (1967) described the isolation of diph-
`theroid organisms from synovial membrane and fluid. Similar results were obtained by
`Hill et al. (1967) who found diphtheroid organisms in eighteen of thirty-seven specimens of
`synovial fluid. However, four of eleven specimens from cases of osteoarthritis also yielded
`similar organisms. In a'n effort to characterize further these organisms, we have investigated
`two selected examples of isolates from cases of rheumatoid arthritis, which were kindly
`provided for us by Drs A. G. S. Hill and J. N. McCormick.
`
`. \
`
`MATERIALS AND METHODS
`Culture of micro-organisms
`Fourteen bacterial cultures from the joints of cases of either rheumatoid arthritis or other
`types of arthritis were cultured on slopes of Loewenstein-Jensen medium. These were all
`provided by Dr A. G. S. Hill from Stoke Mandeville Hospital. ,
`Correspondence: Professor R. G. White, Department of Bacteriology and Immunology, Western In-
`firmary, Glasgow W.1, Scotland. Departmental publication No. 6920.
`
`139
`
`Ex. 1045 - Page 3
`
`

`

`140 (cid:9)
`
`R. G. White and J. Gordon
`
`Electron microscopy
`Bacteria which were taken from Loewenstein-Jensen 5-day slope cultures were suspended
`in 0.15 fvf saline. A carbon coated grid after floating for 30 sec on the surface of this sus-
`pension, was transferred to the surface of 0.15 fvf saline in three successive washing pro-
`cedures, and then on to the surface of distilled water. Subsequently the grid was floated on
`a 1% solution of potassium phosphotungstate at pH 7.2 for 1 min. After removing excess
`stain by blotting paper, the grid was examined in a Siemens Elmiskop IA electron micro-
`scope.
`
`Tests for adjuvant activity by heat-killed bacteria in guinea pigs
`Tests for adjuvant activity of bacterial isolates used a procedure which has been exten-
`sively employed in this laboratory (White et al., 1964). Briefly the method consists in the
`injection of 2 mg of thrice-crystallized ovalbumin as immunogen, in water-in-oil emulsion
`with 200 pg of heat-killed micro-organisms under test in the oil phase. Guinea-pigs were
`each injected into the left hind footpad with 0.2 ml of emulsion composed of saline solution
`of antigen, emulsifying agent (mannide mono-oleate) and mineral oil (Bayol 55) in the
`proportion by volume of 1:1:3. Animals were bled from the heart at 21 days after injection
`and serum prepared from the clotted sample of blood.
`
`Estimation of serum anti-ovalbumin levels
`Serum anti-ovalbumin precipitin levels were estimated by a modification of the Mancini
`single radial diffusion method in which antigen was incorporated into molten 1% oxoid
`ion-agar at a concentration of 100 pg/ml. After solidification a standard volume of anti-
`serum was introduced into wells of 1 mm diameter. The diameter of the resultant precipitin
`rings which developed after 48 hr incubation at room temperature were recorded.
`
`Estimation of serum anti-ovalbumin by complement fixation
`The method used was modified from Price (1949). Fresh guinea-pig serum sensitized
`sheep cells and normal saline were used. The tests were done in tubes using doubling
`dilutions of serum inactivated by heating for 30 min at 56°C. Complement (0.1 ml) con-
`taining 1.25 MHD and 0.1 ml of a solution of ovalbumin (2.5 µg/ml) were used. Sheep red
`cells (2.5% suspension sensitized with 5 MHD of horse amboceptor from Burroughs-
`Wellcome) were added and incubated at 37°C for a further 30 min. Finally the tubes were
`allowed to stand for 1 hr at 4°C before reading, the highest dilution showing any inhibition
`of lysis being taken as the end point. Anticomplementary activity was detected using 0.1 ml
`of a 1:4 dilution of guinea-pig serum together with 0.1 ml of saline instead of ovalbumin and
`1 MHD of complement.
`
`z
`
`Corneal tests
`After anaesthesia of the corneal surface has been established by the installation of 2%
`cocaine hydrochloride, intra-corneal injections were made using a needle size 30 (Imperial
`standard wire gauge) attached to a 1-ml tuberculin syringe. A solution of egg albumin,
`20 mg/ml, was injected in amount sufficient to cause a disk of opacity in the cornea approxi-
`mately 2 mm in diameter. The eyes were examined at 24 and 48 hr, and the extent and
`degree of corneal opacity and the presence or absence of chemosis, recorded. The severity
`of the reaction was graded from 0 to 3.
`
`S
`
`M 0 9
`
`Adjuvant-active micro-organisms in rheumatoid arthritis (cid:9)
`
`141
`
`All the sera from guinea-pigs were collected after killing the animals at the end of the
`3rd week of the experiment.
`
`RESULTS
`
`The two selected isolates (S1 and S2) showed essentially the same cultural characteristics
`in the following investigations.
`Microscopy. Short Gram-positive rods. Old cultures demonstrated a change to cocco-
`
`FIG. 1. Electron micrograph. Negatively-stained preparation of isolate SI showing network
`of filaments on the surface of this micro-organism (diameter 125 A approximately).
`x 134,000
`
`bacillary forms. Young cultures stained by the Ziehl Neelsen method from Loewenstein-
`Jensen solid medium were weakly acid fast, i.e. they were decolorized rapidly with 5%
`sulphuric acid but retained the carbol fuchsin with 1% sulphuric acid. No branching forms
`or spores were seen. Non-motile. Staining by Albert's method failed to reveal volutin
`granules.
`Culture. Aerobe and facultative anaerobe, growing freely on simple media. Colonies were
`1 mm diameter at 24 hr and 5 mm diameter at 5 days. Colony yellow or orange at 5 days
`after access to light. Growth on agar non-adherent, dry, wrinkled and difficult to emulsify.
`Optimum temperature range 25-37°C.
`Biochemical tests. Growth in litmus milk medium failed to produce acid or clot. A range
`of various sugars in peptone medium failed to be fermented. Gelatin failed to be liquefied
`and starch was not hydrolysed.
`
`Ex. 1045 - Page 4
`
`

`

`143
`
`Adjuvant-active micro-organisms in rheumatoid arthritis (cid:9)
`'
`Electron microscopy. The results by negative staining with phosphotungstic acid of Si are
`illustrated in Fig. 1. A network of filaments is ilske. The same appearance was detected on
`repeated isolates from 5-day cultures from Loewenstein-Jensen medium. These appearances
`may be compared with a comparable preparation of Mycobacterium tuberculosis var
`hominis strain H37Rv shown in Fig. 2. In this case, the filaments are rather more definite
`in outline and measure 133 A± 12 A. The filaments of isolate SI (Fig. 1) are slightly more
`slender and were found to measure 125 A±10 A. Fig. 3 is an electron micrograph of a
`TABLE 1. Serum precipitin levels and delayed-type hypersensitivity to ovalbumin in guinea-pigs
`injected with water-in-oil emulsions of ovalbumin with and without test organisms Si and S2
`(Serum antibody determined at 3 weeks and delayed-type hypersensitivity reactions at 19 days)
`
`Diameter of (cid:9)
`Group Animal (cid:9)
`number precipitin ring (cid:9)
`in radial (cid:9)
`diffusion test (cid:9)
`(mm) (cid:9)
`
`Titre of (cid:9)
`complement (cid:9)
`fixation (cid:9)
`(arbitary units (cid:9)
`per ml) (cid:9)
`
`Corneal Skin reaction
`at 48 hr;
`reaction (cid:9)
`diameter of
`at 48 hr (cid:9)
`necrosis:
`diameter of
`erythema
`
`Test
`origin
`SI
`
`Test
`origin
`S2
`
`Neither
`organism
`
`1
`2
`3
`4
`5
`6
`7
`8
`9
`10
`11
`12
`13
`14
`15
`
`6.5
`6-0
`6.5 Mean 6.0
`5.51
`5.5
`3.5
`6.5
`4.5 Mean 5.0
`5.0
`5.5
`2.5
`2.5
`2-5 Mean 2.5
`4.0
`3.0
`
`256
`> 1024
`> 1024 Median > 1024
`anticomplementary
`> 1024
`256
`> 1024
`128 Median 256
`512
`128
`32
`4
`32 Median 32
`512
`
`10:18
`10:20
`11:21
`6:18
`8:17
`5:7
`3:11
`5:12
`13:24
`12:24
`9:18
`9:16
`9:8
`9:20
`10:18
`
`negatively stained preparation of isolate S2, wInch has a similar appearance to that of isolate
`St Filaments again measured 125 A +10 A. \
`Use of the same negative staining techniques on bacteria isolated from 5-day cultures on
`Loewenstein-Jensen medium have failed to demonstrate a comparable filamentous network
`with Corynebacterium diphtheriae var mitis, intermedius or gravis, C. hoffinanni, C. xerosis,
`Listeria monocytogenes and Erysipelothrix rhusiopathiae.
`
`Investigation of activity of isolated bacteria as adjuvants
`Table 1 gives the results of skin and corneal tests sensitization and serum antibody levels
`in guinea-pigs injected with 2 mg ovalbumin in water—oil emulsion with and without added
`200pg SI and S2. Intradermal tests were done with 1 mg ovalbumin in 0 1 nil. The skin
`lesions showed Arthus as well as delayed type reactions and when measured at 48 hr there
`was no significant difference between the reactions of the test groups (injected with Si and
`S2) and the control group. Three of the corneal responses of the five guinea-pigs injected
`
`no. 2. Electron micrograph. Negatively-stained preparation of Mycobacterium tuberculosis
`var hominis H37Rv showing surface network of filaments (diameter 133 A approximately).
`x 134,000
`
`Flo. 3. Electron micrograph. Negatively-stained preparation of isolate S2 showing network of
`filaments on the surface of this micro-organism (diameter 12S A approximately). x 134,000
`
`Ex. 1045 - Page 5
`
`(cid:9)
`(cid:9)
`

`

`144 (cid:9)
`
`R. G. White and J. Gordon
`
`Adjuvant-active micro-organisms in-rheumatoid arthritis (cid:9)
`
`145
`
`with S1 were graded as 2 and all of those injected with S2 were graded as 2 (or 3). Thus both
`Si and S2 induced delayed-type hypersensitivity to ovalbumin. The estimation of precipitin
`levels in 21-day sera showed that guinea-pigs in the experimental groups injected with Si
`and S2 had significantly higher precipitin levels than the controls (average values of 6.0 and
`5.0 as compared with 2.5). Estimations of complement-fixing antibody (Table 1) show that
`guinea-pigs injected with S1 and S2 produced significantly higher levels than the controls
`(medium values of > 1024 and 256 as compared with 32).
`
`these adjuvants stimulate an increase (measured at 3 weeks after injection) of antibody in the
`form of y2-immunoglobulin and increased corneal responses (White, Jenkins & Wilkinson,
`1963). Stimulation of y2-immunoglobulin may be inferred to have taken place as part of the
`responses to organisms S I and S2 since complement-fixation titres to ovalbumin were
`increased (yM antibody to ovalbumin is low or absent at this time after injection). Thus the
`adjuvant effect induced by the micro-organisms S1 and S2 closely resembled that shown by
`peptido-glycolipid components of mycobacteria.
`
`DISCUSSION
`
`REFERENCES
`
`ARAI, M., ISHIKAWA, A. & Horra, E. (1964) Proc. 8th Congr. Japanese Rheumatism Association, Okayama,
`p. 279.
`BARTHOLOMEW, L.E. (1965) Isolation and characterization of mycoplasmas (PPLO) from patients with
`rheumatoid arthritis, systemic lupus erythematosus and Reiter's syndrome. Arthr. and Rheum. 8,
`376.
`CAPLAN, A. (1953) Certain unusual radiological appearances in the chest of coal-miners suffering from
`rheumatoid arthritis. Thorax, 8, 29.
`DUMONDE, D.C. & GLYNN, L.E. (1962) The production of arthritis in rabbits by an immunological reaction
`to fibrin. Brit. J. exp. Path. 43, 373.
`DUTHIE, J.J.R., STEWART, S., ALEXANDER, W.R.M. & DRAYHOFF, R. (1967) Isolation of diphtheroid organisms
`from rheumatoid synovial membrane and fluid. Lancet, i, 142.
`GORDON, R.E. (1966) Some strains in search of a genus—Corynebacterium, Mycobacterium or what? J.
`gen. Microbiol. 43, 329.
`GOUGH, J., RIVERS, D. & SEAL, R.M.E. (1955) Pathological studies of modified pneumoconiosis in coal
`miners with rheumatoid arthritis (Caplan's syndrome). Thorax, 10, 9.
`HILL, A.G.S., MCCORMICK, J.N., GREENBURY, C.L., MORRISS, C.J. & KENNINGDALE, J. (1967) Cultivation of
`micro-organisms from rheumatoid synovia. Ann. rheum. Dis. 26, 566.
`PEARSON, C.M. (1956) Experimental production of arthritis in rats. Ann. rheum. Dis. 15, 379.
`PRICE, I.N.O. (1949) Complement fixation technique: estimation of complement doses. Brit. J. vener. Dis.
`25, 157.
`WHITE, R.G. (1965) The role of peptidoglycolipids of M. tuberculosis and related organisms in immuno-
`logical adjuvance. In Molecular and Cellular Basis of Antibody Formation (Ed. by J. Sterzl), p. 71.
`Czechoslovak Acad. Sci.
`WHITE, R.G., BERNSTOCK, L., JOHNS, R.G.S. & LEDERER, E. (1958) The influence of components of M.
`tuberculosis and other mycobacteria upon antibody pi.oduction to ovalbumin. Immunology, 1, 54.
`WHITE, R.G., JENKINS, G.C. & WILKINSON, P.C. (1963) The production of skin sensitizing antibody in the
`guinea pig. Int. Arch. Allergy, 22, 156.
`WILKINSON, P.C. & WHITE, R.G. (1966). The role of mycobacteria and silica in the immunological response
`of the guinea-pig. Immunology, 11, 229.
`
`There are several cogent reasons for believing that substances with immunological adjuvant
`activity may be concerned in the pathogenesis of rheumatoid arthritis. A polyarthritis
`resembling rheumatoid arthritis has been produced in rats after a single injection of
`Freund's water-in-oil adjuvant (so-called adjuvant disease); Pearson (1956). The myco-
`bacterial component is essential for this process and the rat arthritis can be reproduced by
`injection of mycobacterial peptidoglycolipid in mineral oil. Secondly, another experimental
`procedure productive of arthritis in the rabbit (Dumonde & Glynn, 1962) also makes use of
`Freund-type adjuvant containing mycobacteria for the production of sensitization to
`heterologous or autologous fibrin. Thirdly, the presence of silica particles renders the lung
`vulnerable to a nodular rheumatoid process (Caplan, 1953; Gough, Rivers & Seal, 1955).
`Silica is in many ways a comparable adjuvant to mycobacterial peptidoglycolipid and can
`produce identical immunological effects in the guinea-pig (Wilkinson & White, 1966).
`In this investigation we examined two cultures of micro-organisms (S1 and S2) which had
`been isolated by Drs J. N. McCormick and A. G. S. Hill from the joints of patients with
`rheumatoid arthritis. , These particular organisms were selected for further study since
`investigation of their surface characteristics by negative staining of electron-microscopic
`preparations showed that they both possessed a distinctive network of filaments resembling
`the peptidoglycolipid filaments which are uniformly present on the surface of myco-
`
`racteria (White, 1965) (Figs. 1 and 3). An extensive survey by the authors has shown that
`
`adjuvant-active mycobacteria (of human, bovine, atypical and saprophytic strains) all
`possess a similar surface network of filaments 133 A diameter, as shown in Fig. 2 (Myco-
`bacterium tuberculosis var hominis H37Rv). Adjuvant-active Nocardia rhodochrous and
`Corynebacterium rubrum also possesses a similar surface filamentous network, but adjuvant
`inactive species such as Corynebacterium diphtheriae (var mills, gravis and intermedius)
`as well as C. hofmanni, C. xerosis, Listeria monocytogenes and Erysipelothrix rhusiopathiae
`do not possess comparable surface filaments. Since the C. rubrum referred to here (A.T.C.C.
`14898) has been shown by Gordon (1966) to be a Nocardia organism, it may be the case that
`surface filamentous structures of the type considered here are restricted to Mycobacteria and
`Nocardia species.
`Investigation of the adjuvant activity of orga'nisms S1 and S2 employed a procedure which
`has been used in this laboratory for many years (White et al., 1958). Guinea-pigs injected
`with 200 fig of heat-killed organisms Si and S2 in water-in-oil emulsion with 2 mg oval-
`bumin produced higher serum anti-ovalbumin levels as estimated by precipitin and com-
`plement-fixation tests than controls which received injections which were similar except for
`the lack of micro-organisms and exhibited significantly greater delayed-type hypersensitivity
`in corneal tests. It is characteristic of mycobacteria or mycobacterial peptidoglycolipid that
`...
`
`Ex. 1045 - Page 6
`
`

`

`CLINICAL AND EXPERIMENTAL IMMUNOLOGY
`
`VOLUME 7 . (cid:9)
`
`NUMBER 1 (cid:9)
`
`. (cid:9)
`
`JULY 1970
`
`CONTENTS
`
`F. D. LINDSTROM: Kappa: lambda light chain ratio in IgG eluted from rheumatoid
`arthritis synovium
`
`J. A. AARLI & 0. ToNDER: Antiglobulin consumption test with sera from patients with
`myasthenia gravis (cid:9)
`
`J. A. AARLI: Binding of y-globulin fragments to muscle tissue (cid:9)
`
`MARGARET TURNER WARWICK & PATRICIA HASLAM: Smooth muscle antibody in
`bronchial asthma (cid:9)
`
`M. INouE & G. KLEIN: Reactivity of radioiodinated serum antibody from Burkitt's
`lymphoma and nasopharyngeal carcinoma patients against culture lines derived
`from Burkitt's lymphoma
`
`I. A. MCGREGOR, D. S. RowE, M. E. WILSON & W. Z. BILLEWICZ: Plasma immuno-
`globulin concentrations in an African (Gambian) community in relation to season,
`malaria and other infections and pregnancy
`
`B. M. GREENWOOD, ERINA M. HERRICK & E. J. HOLBOROW: Speckled antinuclear
`factor in African sera
`
`G. VIRELLA & MARIA F. LOPES-VIRELLA: Effects of therapeutically useful thiols (DL-
`penicillamine and a-mercaptopropionylglycine) on immunoglobulins
`
`1 I
`
`23
`
`31
`
`39
`
`5L
`
`H. W. KRETH, G. THIESSEN & H. DEICHER: The effect of antigen given intravenously
`on specific antigen-sensitive lymphocytes of peripheral blood in man (cid:9)
`
`109
`
`A. PASTERNACK & E. LINDER: Renal tubular acidosis : an immunopathological study on
`four patients (cid:9)
`
`115
`
`CASE REPORT
`
`M. I. DRURY, DEBORAH M. KEELAN, F. J. TIMONEY & W. J. IRVINE: Juvenile familial
`endocrinopathy (cid:9)
`
`125
`
`BLACKWELL SCIENTIFIC PUBLICATIONS
`OXFORD AND EDINBURGH
`
`Printed and bound in Great Britain by Alden & Mowbray Ltd
`at the Alden Press, Oxford
`
`CLINICAL AND
`EXPERIMENTAL
`IMMUNOLOGY
`
`EDITOR W. J. IRVINE
`
`Blackwell Scientific Publications
`Annual Subscription £18 ($60.00) post free
`Single issues at 35s ($6.00) net
`
`VOL 7 NO 2 AUGUST 1970
`An official journal of the
`British Society for Immunology
`
`Ex. 1045 - Page 7
`
`

`

`CLINICAL AND EXPERIMENTAL IMMUNOLOGY
`NUMBER 2 . AUGUST 1970
`
`VOLUME 7 (cid:9)
`
`CONTENTS
`
`R. M. STROUD, K. NAGAKI, R. J. PICKERING, H. GEWURZ, R. A. GOOD & M. D.
`COOPER: Sub-units of the first complement component in immunologic deficiency
`syndromes : independence of Cls and Clq
`R. G. WHITE & J. GORDON: Adjuvant-activity of `diphtheroid' organisms isolated
`from the joints of cases of rheumatoid arthritis
`J. B. ZABRISKIE, K. C. Hsu & B. C. SEEGAL: Heart-reactive antibody associated with
`rheumatic fever: characterization and diagnostic significance
`A. DE SCHRYVER, G. KLEIN & G. DE THE: Surface antigens On lymphoblastoid cells
`derived from nasopharyngeal carcinoma
`G. WICK, E. WITEBSKY, J. H. KITE, JR. & E. H. BEUTNER: Immunofluorescent studies
`of the thyroid auto-antibodies in chickens of the Obese strain (OS)
`G. WICK: The effect of bursectomy, thymectomy and X-irradiation on the incidence
`of precipitating liver and kidney auto-antibodies in chickens of the Obese strain (OS)
`R. P. MouroN, J. W. STOOP, R. E. BALLIEUX & N. A. J. MuL: Pneumococcal anti-
`bodies in IgA of serum and external secretions
`G. KRONVALL & H. GEWURZ: Activation and inhibition of IgG mediated complement
`fixation by staphylococcal protein A
`R. OHNO, J. E. HARRIS & E. M. HERSH: L-asparaginase: suppression of the immune
`response of mice to sheep red blood cells
`P. TH. A. ScHELLEKENs & V. P. EIJSVOOGEL: Lymphocyte transformation in vitro.
`III. Mechanism of stimulation in the mixed lymphocyte culture
`A. S. COULSON, LUCY J. SUMMERS, KERSTIN LINDAHL-KIESSLING, D. TUCKER & K.
`HELLMANN: The effect of two soluble thalidomide derivatives on lymphocyte
`stimulation
`R. A. ALLARDYCE & K. JAMES: Studies on the effect of phytohaemagglutinin and
`anti-lymphocytic globulin on the primary humoral response to sheep erythrocytes
`and bovine serum albumin in Hooded and Wistar rats
`0. J. MELLBYE & J. B. NATVIG: Experimentally produced antibodies to the pepsin
`site of IgG due to untreated autologous IgG in immune complexes
`DIANE J. YASHPHE & D. W. WEISS: Modulation of the immune response by a
`methanol-insoluble fraction of attenuated tubercle bacilli (BCG). I. Primary and
`secondary responses to sheep red blood cells and T2 phage
`M. MAJER & F. LINK: Studies on the non-neutralizable fraction of vaccinia virus
`
`133
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`139
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`147
`
`161
`
`173
`
`187
`
`201
`
`211
`
`221
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`229
`
`241
`
`249
`
`257
`
`269
`283
`
`SHORT COMMUNICATION
`
`P. K. JOHNSON & J. M. YODER: Unexpected extent of immunochemical cross-reac-
`tions between rabbit and human serum proteins
`BOOK REVIEWS; BOOKS RECEIVED (cid:9)
`
`293
`299
`
`BLACKWELL SCIENTIFIC PUBLICATIONS
`OXFORD -AND_ EDINBURGH
`
`Printed and bound in Great Britain by Alden & Mowbray Ltd
`at the Alden Press, Oxford
`
`55
`
`„ (cid:9)
`
`'
`
`• 1
`
`0
`
`CLINICAL AND
`EXPERIMENTAL
`IMMUNOLOGY
`
`EDITOR W. J. IRVINE
`
`Blackwell Scientific Publications
`Annual Subscription £18 ($60.00) post free
`Single issues at 35s ($6.00) net
`
`VOL 7 NO 3 SEPTEMBER 1970
`An official journal of the
`British Society for Immunology
`
`rs
`
`o •
`
`I
`
`5
`
`rte
`
`Ex. 1045 - Page 8
`
`