Proceedings of the northeastern regional meeting of the American rheumatism association.код для вставкиСкачать
283 Proceedings of the Northeastern Regional Meeting of the American Rheumatism Association December 5-6, 1974 Newton, Massachusetts Abstracts of Papers Presented Relationship Between Cartilage “Healing” and Applied Stress Howard G. Parker and Eric L. Radin, Boston, Massachusetts Although degenerative joint disease has been characterized as a proliferative response that increases metabolic activity in the tissues, it is generally accepted that this proliferative response is never functionally successful and that once begun joint degeneration is unrelenting. Experimental attempts to heal very large, full thickness defects in articular cartilage have been unsuccessful. In order to examine the relative role of intraarticular pressure on the healing of grossly damaged articular surfaces, the articular cartilage was completely denuded from the weight-bearing areas of both sides of one hip joint of 70 adult cats. These animals were followed for periods up to 30 months. Onehalf the animals also had modified “hanging hip” procedures in conjunction with the cartilage removal. The control animals stopped limping between postoperative months 3 and 6, but the limp reappeared in this group by month 12 and persisted. Limping in the tenotomized (experimental) group disappeared between months 6 and 12 and did not reoccur. Hypertrophic lips and spurs were consistently found in the control animals at month 24. Most of the animals in this group had a limited range of motion of the hip, and the capsule about the joint was thickened. Frank fibrous ankylosis occurred in 2 control animals. The experimental animals had hip joints that remained congruous and were free of lips and spurs. Microscopically, fibrous tissue was present as early as 1 month in the articular defects in all animals but was clearly more exuberant in the tenotomized animals. After 1 year histologic sections taken through the weight-bearing portions of the heads and acetabulae of the tenotomized group showed fibrocartilage and what appeared to represent transformation of small focal areas of fibrocartilage into an intermediate stage, more hyaline than fibrous. T h e chondroid nature of this tissue was confirmed by Safranin-0 staining. There was no indication that the small rim of cartilage left in the periphery of the joint contributed to the healing. T h e hips of the animals with the multiple tenotomies went on to fibrocartilaginous and chondroid healing of their articular surfaces and clinically did well. Judged by all criteria the tenotomized hips were consistently functionally excellent at month 30 as compared to those of the control animals who pathologically and clinically had degenerative arthrosis. Small Airway Disease in Scleroderma M. Guttadauria, D. Saroja, P. Gopinathan, H. Ellman, G. Emmanuel, H. Lyons, H. Diamond, and D. Kaplan, Brooklyn, New York Pulmonary Function Tests (PFTs) were performed in 46 patients with Progressive Systemic Sclerosis (PSS), 35 women and 11 men. Parameters measured were vital capacity (VC), total lung capacity (TLC), residual volume (RV), diffusion capacity of the lung for carbon monoxide (DLCO), midmaximum expiratory Row rate-FEF25%-,5% (MMFR). and forced expiratory volume in 1 second (FEV,). The findings were not those that have been considered typical of PSS. Twenty-nine patients (63%) were found to have small airway disease (SAD) as manifested by an abnormally low MMFR (1.5 f 0.59 liter/sec; 50.5 f 16.8% Pred). Thirteen of these patients (28%) had frank obstructive lung disease as evidenced by the finding of a low FEV, (67.83 7.05% of FVC). Smokers comprised only 7/29 (24%) of + Arthritis and Rheumatism, Vol. 18, No. 3 (May-June 1975) the patients with a low MMFR and 14/46 (30%) of the total. An additional 13 patients (23%) had abnormalities suggestive of SAD. These patients had an elevated RV (2.18 -t 0.71 liter; 176.5 -C 29.9% Pred) with normal MMRF and FEV,. These patients are considered to have SAD because of the close relationship that has been shown to exist between RV and closing volume. The latter measurement was not made in our patients. Radiographic evidence of lung cysts was not present. Thus 42 of 46 patients (91%) manifested some evidence of SAD. Only 3 patients (7%) had PFTs consistent with classic restrictive lung disease-decreased RV, TLC, and VC-without evidence of SAD. One patient had normal PFTs. Obstructive lung disease had been considered not only unusual in PSS but also a manifestation of advanced 284 AF2A ABSTRACTS disease when it occurs. Twenty-two of 42 (52%) of our patients with SAD had PSS for less than 2 years. Twentythree of 42 (55%) had no dyspnea or other pulmonary symp toms. Twenty-six of 42 (62%) had normal chest film. T e n of 42 (24%) had normal chest ratio-gram, no dyspnea, and disease of less than 2 years duration. Thus, the finding of SAD is not restricted to patients with advanced disease. These data suggest that involvement of the small airways is a frequent finding in PSS. Detection of Antibody to Native DNA(N-DNA):Increased Specificity for Active SLE by Using a Synthetic N-DNA Antigen Free of Contaminating Single-StrandedDNA (ss-DNA) Charles R. Steinman, Utis Deesomchok, and Harry Spiera, Mount Sinai School of Medicine, New York Contamination with ss-DNA of most or all natural N-DNA antigens used for measuring antibody to N-DNA may account for recently reported detection of such antibody in non-SLE states. Some denaturation may be virtually unavoidable during purification and handling of natural N-DNA. Hence it is proposed that a synthetic, self-complementary, alternating copolymer of deoxyadenylate and deoxythymidylate (dAT) be substituted for natural N-DNA (KB DNA) in such assays. This substance would, on theoretical grounds, be expected to be free of ss-DNA and other cellular antigens and should carry the same antigenic determinants as natural N-DNA. Immunochemical comparison of dAT with KB DNA was carried out. T h e above expectations were confirmed. All natural N-DNA preparations examined, including those repurified to remove ss-DNA by MAK chromatography or by treatment with an ss-DNA- specific nuclease, still contained detectable ss-DNA. Binding of dAT in the ammonium sulfate assay for anti-N-DNA by sera from patients with SLE and other diseases as well as from normals was compared with binding of KB DNA. dAT binding appeared to be more specific for SLE, particularly for active SLE nephritis. Active neurologic disease was not associated with increased dAT binding. T h e percent dAT binding, when elevated, paralleled disease activity in the same manner as did KB binding. In at least 1 patient the only clinical evidence of diffuse proliferative nephritis was the elevated dAT binding. Limited data comparing renal biopsies with coincidental dAT binding will be presented. It is concluded that use of d A T to detect anti-N-DNA antibody offers theoretical and practical advantages over use of natural N-DNA including greater specificity, better reproducibility, and lower cost. Antibodies to DSDNA in Patients with Rheumatic Disease Other than Systemic Lupus (SLE) C. Bell and P. H. Schur, Robert B. Brigham Hospital, Harvard Medical School, Boston, Massachusetts Antibodies to double-stranded DNA were detected by counterimmunoelectrophoresis and measured by radioimmunoassay in sera from patients with certain connective tissue disorders other than SLE. Precipitins to DSDNA were found in 40 of 448 sera from patients with rheumatoid arthritis (RA) and in 25 of 162 sera from patients with juvenile rheumatoid arthritis (JRA), as well as 5 sera from patients without RA, JRA, or SLE. Sera and additional clinical information were available on 28 patients who could then be divided into four groups, based on the number of criteria each had for RA and SLE. Group I: 5 patients had five or more criteria for RA and four or more for SLE; Group 11: 13 patients had five or more criteria for RA and less than four criteria for SLE; Group 111: 5 patients with JRA; and Group IV: 5 patients had less than five criteria for RA and less than four criteria for SLE. Patients with RA and anti-DNA antibodies had more severe disease, particularly extraarticular manifestations, than did RA patients without anti-DNA antibodies. Nephritis and hypocomplementemia were observed in 4 of 5 patients in Group I. Based on these observations, it was concluded that the presence of anti-DSDNA should not be used as a sole criterion in establishing the diagnosis of SLE. Various Types of Antilymphocyte Antibodies in Patients with Systemic Lupus Erythematosus (SLE)and Their Relationship with Lymphopenia John B. Winfield, Robert J. Winchester, and Henry J. Kunkel, Rockefeller University, New York, New York A 12-month prospective study of 30 patients was performed to define the relationship of antilymphocyte antibody Ig class and thermal amplitude of binding with lymphopenia. Sera given high in vitro lympholysis at 25°C were highly correlated with lymphopenia in paired blood specimens (r = 0.66, P 0.0001 by linear regression analy- < 285 ARA ABSTRACTS sis). A variety of different antibodies were found. IgM antibodies predominated and appeared to be the only type active in in vitro cytotoxicity. Different thermal amplitudes could be demonstrated by indirect immunofluorescence and cytotoxicity carried out at 15"-37"C for IgM antibodies and by immunofluorescence for the much less frequent IgG types, although marked cold enhancement of binding (4'C) was universally present. Representative clinical courses regarding these different types of antibodies will be presented. The data suggest that lymphopenia may be directly related to cold-reactive IgM antibodies, particularly those exhibiting a relatively higher thermal amplitude of binding. Twin Studies in Systemic Lupus Erythematosus (SLE) Sidney R. Block, John B. Winfield, Michael D. Lockshin, William A. D'Angelo, Marc E. Weksler, Marilena Fotino, and Charles L. Christian, New York, New York Clinical and serologic characteristics and certain cellmediated immune functions were studied in 12 sets of twins (7 monozygotic, by erythrocyte and leukocyte antigens) of which one or both twins had SLE. Four of 7 monozygotic pairs were concordant for overt SLE (ARA criteria); 5 for ANF, 6 for hypergammaglobulinemia. (Similar data of 17 previously reported sets will be reviewed.) In other firstdegree relatives the prevalence of ANF was 28 percent and hypergammaglobulinemia 33 percent. Lymphocytes of 6 monozygotic pairs, including the 3 clinically discordant sets, failed to demonstrate stimulation in mixed leukocyte cultures (twin versus twin) but stimulated and were stimulated by unrelated leukocytes. Sera of 2 of 3 SLE twins (whose lymphocytes exhibited poor mitogen-induced blastogenesis) did not suppress the normal responses of their discordant twin's lymphocytes; nor did the SLE twin's leukocytes recover in the well twin's serum. Cold-reactive IgM lymphocytotoxic antibodies were present in the sera of most SLE twins, but only in 1 well twin. Each lymphocytotoxic serum was equally reactive with normal and SLE peripheral blood lymphocytes. The data suggest a strong genetic component in the pathogenesis of SLE and an acquired defect in leukocyte function. Evidence for neoantigens on SLE lymphocytes, foreign to the well twin of clinically discordant monozygotic pairs, was not obtained. Distribution of HLA Antigens in Patients with Juvenile Rheumatoid Arthritis (JRA) D. Gibson, P. H. Schur, C. B. Carpenter, J. S. Stillman, Robert B. Brigham Hospital, Peter Bent Brigham Hospital, Harvard Medical School, Boston, Massachusetts The recent demonstration of an association between certain HL-A phenotypes and JRA stimulated us to reexamine whether a similar relationship could be demonstrated in our patients. Thirty HL-A antigens were examined in 126 normal Caucasian controls and 122 patients with JRA. W-17 was found in 7% of JRA patients and in 0.01); HL-A, was found in 18% of controls (a' 6.72, P 597" of JRA patients and in 41% of the controls (a' 7.11, P 0.01). Although there was no evident difference in the frequency of these phenotypes in JRA patients with or without nodules, lymphadenopathy, serositis, uveitis, ANA, < < rheumatoid factors, type of onset, or a family history of arthritis, HL-A was found in 69% of JRA patients with hepatomegaly, as compared to 23% of JRA patients with0.005). W27 was found in out hepatomegaly (a* 9.01, P 16% of JRA patients and in 6% of the normal controls (a' 3.14, P 0.1). Of those JRA patients with W27 only 1 of 13 had radiologic evidence of sacroilitis. Although trends are noted between HL-A phenotypes and aspects of JRA, these studies fail to confirm the observations of others reporting a relationship of W27 and JRA. < < Connective Tissue Proteins Synthesized by Scleroderma Fibroblasts in Cell Culture Roy Fleischmann and E. Carwile LeRoy, College of Physicians and Surgeons of Columbia University, New York, New York Soluble precursors of collagen in the culture medium of fibroblasts obtained by skin biopsy from 5 patients with scleroderma (SD) were characterized by SDS-acrylamide gel electrophoresis and compared to the medium of normal human fibroblasts (NL) matched for age, sex, race, and biopsy site. By using the parameters of molecular size, patterns of dual isotope incorporation (3H tryptophan and 14C proline), sensitivity to pepsin and bacterial collagenase, and of reduction and alkylation, both SD and NL medium contained newly synthesized proteins that were character- 286 ARA ABSTRACTS ized as polymers of procollagen and monomers of collagen and procollagen. Electrophoretic patterns of NL and SD medium were qualitatively similar. T h e ratio of collagenous protein synthesis as demonstrated by collagenase sensitivity was 1.8 (SD:NL, five culture pairs); using hydroxyproline analysis the ratio was 2.2 (collagen/flask) and 1.3 (collagen/ cell) in three culture pairs. A previously unrecognized, noncollagenous protein (or proteins) in the gel region of procollagen polymers was characterized as pepsin sensitive, bacterial collagenase resistant, nonreducible, and possibly glycoprotein or glycosaminoglycan in nature. These noncolla- gcnous materials represent a substantial proportion of the NL and SD fibroblasts synthetic output (s 50% of incorporated label) and can be mistaken for procollagen polymers by electrophoretic mobility and isotope incorporation. These data confirm the initial impression of increased collagen synthesis by SD fibroblasts in vitro and are consistent with an orderly assembly and maturation from procollagen to collagen in SD fibroblasts as shown with NL fibroblasts. Increased collagen synthesis by SD fibroblasts is consistent with an activated or incompletely regulated state of the SD fibroblast in vitro. Release of RNA Virus from Human Rheumatoid Synovial Fibroblast-Mouse Fibroblast Hybrids Olaniyi Kehinde and Robert Poss, hlassachusetts Institute of Technology, Harvard Medical School, and Robert Breck Brigham Hospital Somatic cell hybridization has proved a useful tool for studying mechanisms of genetic control. Using these techniques, cell functions ordinarily not expressed can be “turned on”; conversely, a gene usually expressed in one of the parent cells may not be expressed in the hybrid. We have utilized a human-mouse hybrid to study certain properties of the rheumatoid synovial fibroblast. Rheumatoid synovial tissue obtained at the time of surgery was grown in tissue culture until confluent monolayers were established. These cells were then cocultivated with mouse fibroblasts in the presence of inactivated Sendai virus. Clones were selected after growing the cell populations in selective HAT media. Six clones have been isolated, grown in quantity, and carried in HAT media. Each clone differs in growth characteristics and in karyotype. We have tested these clones for their ability to express RNA tumor virus; particularly, we have studied the contribution of the human rheumatoid synovial cell genome to virus release from the mouse-human hybrid. Although a viral etiology of rheumatoid arthritis is suspected, no virus has been previously isolated from rheumatoid tissues. T h e implications of our finding that RNA tumor virus is released from human rheumatoid-mouse fibroblast hybrids are discussed. Desensitized Human Synoviocytes: A Model for Investigating the Interaction Between Prostaglandins and Other Hormones D. S. Newcombe, C. P. Ciosek, Jr., J. V. Fahey, and R. W. Ortel Human synoviocytes in culture exposed to prostaglandin (PG) El and E, demonstrate a dramatic increment in intracellular cyclic AMP (ICAMP) concentration. Synoviocytes, after preincubation in serum-free media, have an ICAMP concentration (pmoles per milligram of protein) of 79.3 (+ 17.5), whereas synoviocytes exposed to 3 pg per milliliter of PGE, or PGE,, for 15 minutes, have concentrations of 1890.5 (& 248) and 393.5 (+ 9.6), respectively. PGA,, PGA,, PGB,, PGB,, and PGF,, at concentrations u p to 10 r g per milliliter do not significantly increase synoviocyte ICAMP concentrations; 7-oxa-13-prostynoic acid inhibits the PGE, response. A maximum ICAMP response to PGE, or PGE, occurs in 15 minutes and returns to approximately basal ICAMP levels 2 to 4 hours later. PGE,or PGE,-stimulated ( 3pg per milliliter) synoviocytes re- spond minimally to rechallenge with PGE, or PGE2 (desensitization) after ICAMP concentrations have returned to basal levels. PGE,-containing media from desensitized cells transferred to untreated synoviocytes elicit an ICAMP response similar to primary PGE, treatment. Desensitized synoviocytes when retested with PGE, remain unresponsive for at least 72 hours after initial PGE, treatment. Desensitized synoviocytes require 36 hours in the presence of PGE,free media to regenerate full PGE, responsiveness. PGE,desensitized synoviocytes are also unresponsive to PGE, and epinephrine. Epinephrine (1.8 rcg per milliliter) alone results in a doubling of ICAMP as compared to untreated synoviocytes. Massive doses of PGE, (30 pg per milliliter) added to PGE,-desensitized synoviocytes are unable to overcome the desensitization. ARA ABSTRACTS 287 Bence Jones Protein (BJP) from a Patient with Primary Amyloidosis Identical to the Amyloid Fibril Paul J. Block, Martha Skinner, Merrill D. Benson, and Alan S. Cohen Serum M-components may be found in patients with primary amyloidosis. We recently studied such a patient (AC 72-168) with an IgG M-component who had a nephrotic syndrome and ascites. An M-component was also demonstrated in the peritoneal fluid and found by agarose electrophoresis to be identical with that of her serum. The peritoneal fluid M-component was extracted by ammonium sulfate precipitation and isolated by starch block electrophoresis. The protein was reduced, alkylated, and chromatographed on Sephadex G-100 into its heavy- and lightchain components. The resulting light chain reacted by immunodiffusion with anti-kappa BJP. Amyloid fibrils were isolated from the patient’s liver by exhaustive saline washes, denatured in 5 M guanidine, and fractionated by Sepharose 6B chromatography. The major retarded fraction amounted to approximately 70% of the amyloid fibrils. Amino acid sequence determination was performed on both the isolated light chain from the peritoneal fluid to 30 residues and on the amyloid fibril protein from the patient’s liver to 18 residues. Both began as Asp-Ile-GlnMet-Thr and were identical throughout to one another and to a typical Kappa I type BJB. These results lend support to the hypothesis that light chains of circulating monoclonal immunoglobulins may be precursors of amyloid fibril protein in primary amyloidosis. However, this study cannot rule out the possibility that the circulating immunoglobulins and the amyloid fibrils are produced by separate but similar mechanisms. In Vivo Mechanisms of BCG Immunosuppression R. 1. L. Sutherland, M. A. Spadaro, and F. Quagliata, Rheumatic Disease Study Group, New York University School of Medicine, New York, New York We reported (J Rheumatol 1:26, 1974 [suppl 13) the suppression of adjuvant disease by BCG. I n clarifying these mechanisms, we noted the following. BCG given to rats with florid disease significantly suppressed the arthritis within hours. Delayed hypersensitivity to PPD in rats treated with BCG either before or after the adjuvant injection was positive. BCG also suppressed disease induced by Corynebacterium rubrum instead of M tuberculosis in the adjuvant mixture. When SXr-labeled thoracic duct cells from normal and BCG-treated rats were transferred into syngeneic normal, BCG-treated, adjuvant-injected, and BCG-treated and adjuvant-injected recipients, a marked increase in counts (P 0.001) was found in the thymi of < ERRATUM Levinson DJ, Silcox DC, Rippon JW, and Thomsen S: Septic Arthritis Due to Nonencapsulated C q p t o coccus neoformans with Coexisting Sarcoidosis. Arthritis Rheum 17:1037-1047,1974 rats receiving BCG and/or adjuvant. Concurrently reduced bone marrow localization was noted. Blood, spleen, and liver counts were unchanged. Nodes draining the site of adjuvant injection showed higher counts. Increased homing in thymi of rats receiving BCG was also noted when cells from BCG-treated donors were used. The altered homing pattern probably reflects a structural modification of the plasma membrane of these cells. Our results exclude tolerance, paralysis, antigenic competition, or cross-reactivity as major factors in the suppression and, with our previous data, imply that BCG influences the activity of suppressor T cells, perhaps working in a direct pharmacologic fashion. On page 1043 of the above article the illustration labeled Fig 6 Lower should instead appear above the caption for Fig 7 on page 1044. The illustration on page 1044 is that for Fig 6 Lower.