Anti-collagen Antibodies Research Articles

Antibody formation against human collagen and C1q in response to a bovine collagen implant

The humoral immune response to commercially available bovine collagen implants (Zyderm, Zyplast) is characterized in a 45-year-old female patient. Circulating anti-collagen antibodies were detected after eight injections of Zyderm and after two injections of Zyplast given during a period of 3 years. The specificity of these antibodies for bovine and human collagens as well as for the collagen-like region of C1q (a subcomponent of the first component of complement), was investigated by affinity chromatography. Serum levels of anti-collagen and anti-C1q antibodies were measured using ELISA. High levels of antibodies to bovine collagens, showing a strong cross-reactivity with human collagen type III were detected in the patient's serum. Only weak cross-reactivity with human collagen type I and IV and no reactivity with type II were observed. In addition, these antibodies specifically cross-reacted with the collagen-like region of C1q. The antibody levels decreased continuously and disappeared 1 year after cessation of treatment. These results demonstrate for the first time the formation of autoantibodies upon treatment with a bovine collagen implant. Although antibodies to collagens and C1q have been found in various autoimmune diseases, neither adverse reactions to the bovine collagen implant nor any other clinical symptoms were observed in association with the described antibody response.

Lipoid proteinosis-new immunopathological observations

Lipoid proteinosis is described in a family of Scottish descent. Chromosome studies performed on five members were normal. Direct immunofluorescence studies performed on skin with anti-collagen antibodies showed an abnormal distribution of collagen within the skin. No abnormality was detected in DNA extracted from lymphocytes taken from five family members, when probed with gene probes to collagen types alpha 1 (I), alpha 2 (I), alpha 1 (III) and alpha 2 (V).

Immune response after collagen injection for laryngeal rehabilitation: development of an enzyme-linked immunosorbent assay

The appearance of circulating antibodies directed against bovine collagen has been observed in dermatology after collagen injections with or without clinical reactions. Knowing the serologically positive patients among the clinically negatives ones is important if a second collagen injection should be planned. In these cases, skin and blood tests have to be carefully checked to confirm reactivity. In connection with this, we have developed an enzyme-linked immunosorbent assay (ELISA) for determining the response to GAX collagen injections. Such injections induce a very low rate of anti-collagen antibodies. However, without having recourse to a positive control permitting statistical analysis of data, no conclusions can be made about significant positivity. In the present investigation of 12 patients following GAX collagen injections into the larynx, 1 was found to be borderline and 1 positive (17%). The positive control was made up with 20 ng/ml anti-type III collagen antibodies, which was purified and tested against type III bovine and human collagen. The present study showed that the systematic detection of antinuclear or anti-DNA antibodies after GAX collagen injection is not useful. However, the anti-collagen positivity of two control samples from 3 patients already positive for antinuclear antibodies demonstrates that bovine collagen injections are contraindicated in auto-immune disease.

A monoclonal antiidiotypic antibody with rheumatoid factor activity defines a cross-reactive idiotope on murine anticollagen antibodies.

A syngeneic antiidiotypic mAb, C1C3, was characterized as to its binding to monoclonal anti-collagen II (-CII) auto-antibodies reactive with different epitopes of the native CII molecule. Both by direct binding and by inhibition ELISA studies, the anti-idiotypic antibody was shown to react with a cross-reactive idiotope present on Fab fragments of most, but not all, tested anti-CII mAb, whereas the binding to Fab fragments from normal mouse IgG was low. As previously described, C1C3 bound to isolated Fc fragments from normal mouse IgG. The binding to intact normal mouse IgG was, however, weak, and only isolated Fc-gamma fragments, not intact IgG, competed efficiently with Fab fragments of anti-CII antibodies for binding to the antiidiotypic antibody. The antibody was shown to self-associate, i.e., to behave similarly to certain IgG rheumatoid factors obtained from patients with rheumatoid arthritis. The presented data indicate that the described anti-anti-CII mAb may be representative of antibodies involved in the physiologic regulation of autoimmunity to CII and, consequently, may be used as a tool for further studies on idiotypic regulation in CII-induced arthritis.

Serum IgE anti-cartilage collagen antibodies in rheumatoid patients

The presence of circulating IgG, IgA and IgM antibodies to native cartilage collagens in some patients with rheumatoid arthritis (RA) suggests that an autoimmune response to cartilage collagens may be involved in the pathogenesis of RA. However, the relevance of such antibodies to the pathological process remains unclear, and it is likely that many humoral and cellular derived factors combined to trigger events leading to the chronicity of the rheumatoid lesion. Since histological and biochemical studies have suggested the involvement of mast cells in the rheumatoid joint, we have studied the frequency of IgE antibodies directed against the cartilage collagens type II, IX and XI in patients with active rheumatoid disease. Of the 91 patients' sera tested, 32 had significant levels of IgE anti-cartilage collagen antibodies when compared with non-arthritic controls. Total serum IgE levels did not correlate with the presence of IgE anti-collagen antibodies, nor were any patients positive for IgE antibodies to fibronectin, a widely distributed extracellular matrix component. These results are consistent with an allergic type I hypersensitivity reaction to cartilage antigens in RA involving mast cell and basophil degranulation.

Immune response after collagen injection for laryngeal rehabilitation: development of an enzyme-linked immunosorbent assay

The appearance of circulating antibodies directed against bovine collagen has been observed in dermatology after collagen injections with or without clinical reactions. Knowing the serologically positive patients among the clinically negatives ones is important if a second collagen injection should be planned. In these cases, skin and blood tests have to be carefully checked to confirm reactivity. In connection with this, we have developed an enzyme-linked immunosorbent assay (ELISA) for determining the response to GAX collagen injections. Such injections induce a very low rate of anti-collagen antibodies. However, without having recourse to a positive control permitting statistical analysis of data, no conclusions can be made about significant positivity. In the present investigation of 12 patients following GAX collagen injections into the larynx, 1 was found to be borderline and 1 positive (17%). The positive control was made up with 20 ng/ml anti-type III collagen antibodies, which was purified and tested against type III bovine and human collagen. The present study showed that the systematic detection of antinuclear or anti-DNA antibodies after GAX collagen injection is not useful. However, the anti-collagen positivity of two control samples from 3 patients already positive for antinuclear antibodies demonstrates that bovine collagen injections are contraindicated in auto-immune disease.

Possible formation of cutaneous amyloid from degenerative collagen fibers. Ultrastructural collagen changes and the immunoreactivity of cutaneous amyloidosis employing anti-type I, III, IV, V collagen antibodies.

The origins of primary cutaneous amyloid have been investigated and several possibilities are proposed. We investigated here several cases by electron microscopy and found in all cases characteristic filamentous changes which were identical with amyloid structures in the collagen bundles. These observations suggested a possible pathway of cutaneous amyloid formation from degenerative collagen fibers. Further studies were performed using immunoelectron microscopical and immunohistochemical methods employing a panel of anti-collagen antibodies to examine the reactivity of amyloid to them. Our results were much more suggestive of the collagen origin of this kind of amyloid.

Immune response to collagen impregnated Dacron double velour grafts for aortic and aorto-femoral reconstructions

This study presents 20 patients, randomised to receive either a collagen-treated or an ordinary Dacron graft for aortic reconstructions, and the results of a skin-prick test, blood parameters and ELISA for anti-collagen antibodies as well as NMR pictures during a 6 week follow-up period. Forty per cent (4/11) of those receiving a collagen impregnated graft had a significantly increased titre of antibodies and NMR revealed in two out of 11 patients either a slightly increased amount of fluid or fibrosis around the graft, both collagen impregnated. No differences were found between the graft groups concerning body temperature and leucocyte or platelet counts. The skin-prick test for collagen was negative in all cases.

IgE-containing immune complexes in synovial fluid of patients with rheumatoid arthritis

The prevalence and composition of IgE-containing immune complexes in paired synovial fluid and serum of 42 patients with classical or definite rheumatoid arthritis were studied. IgE-containing immune complexes were found in 15/42 synovial fluids; 15 sera were also positive. The correlation between serum and synovial fluid complexed IgE levels was high (r = 0.77). The mean ratio of synovial fluid/serum levels was 1.96, i.e. significantly higher than 0.33, the synovial fluid/serum ratio for alpha-2-macroglobulin (molecular weight 820 kD), which was taken as high molecular weight control protein (p less than 0.0001). Apart from IgE in immune complex form, monomeric IgE was also significantly higher in synovial fluid compared to serum (ratio = 2.94). Other constituents which could be found in the immune complexes, i.e. anti-IgE antibodies, rheumatoid factors and anticollagen antibodies, were also higher in synovial fluid than predicted. Our results suggest intra-articular production of IgE-containing complexes in the synovial fluid, in addition to possible exudation of the complexes from the serum. These findings provide further evidence for the role of IgE-containing immune complexes in rheumatoid synovitis.

Collagen and cutaneous amyloidoses

A relationship between cutaneous amyloid deposits and collagen fibers has often been described. 1–8 Although this relationship may be important in the development of cutaneous amyloidoses, little attention has been focused on changes in collagen itself, which may be more directly relevant. In 1965, Hashimoto et al. 1 first reported the characteristic collagen changes in a case of lichen amyloidosus and showed filamentous changes of these fibers in the vicinity of the amyloid masses. They attributed this to collagen synthesis from amyloid-like thin filaments, rather than to degeneration of collagen which might have resulted in amyloid production. Some additional short reports of collagen alterations in primary cutaneous amyloidosis have been published, 8–10 but no thorough investigation of this subject has been undertaken. In 1981, we reported a case of macular amyloidosis in which amyloid was thought to be derived from degenerated collagen fibers. 11 In our case, many collagen bundles showed unusual filamentous changes which were sometimes observed to affect whole collagen islands. The altered collagen fibers were very similar to amyloid filaments in ultrastructural appearance. Although a similar report has been added, 12 the total number of cases remains very small, and biochemical or immunologic analysis of the degenerated material has not been done. Therefore, we have examined several cases of macular and lichenoid amyloidoses by electron microscopy, and have investigated antigenic identity between the degenerated material and amyloid employing specific anticollagen (type I, III, IV, V) antibodies by immunologic methods.

Studies on the devlopment of autoantibodies in chickens following experimental reovirus infection

Sera from chickens after experimental oral or foot-pad infection with an avian arthrotropic reovirus were examined for the presence of autoantibodies. Antinuclear antibodies and low levels of anti-collagen antibodies were observed in a proportion of samples, but rheumatoid factor was not detected.

Effects of indomethacin, cyclosporin, cyclophosphamide, and placebo on collagen-induced arthritis of mice

The long term effects of indomethacin, cyclosporin, cyclophosphamide, and placebo on collagen-induced arthritis in mice were tested under two different treatment protocols. A prophylactic experiment examined the effects of the daily drug administration for 180 days beginning one day before the first collagen injection. Under this dosage schedule, cyclophosphamide and cyclosporin decreased the severity of arthritis, while indomethacin did not. A therapeutic protocol examined the effects of these same drugs when daily administration was delayed until the animals had active disease at 78 days after the first collagen injection. Under this protocol, all three drugs reduced the progression of disease. In both protocols, the most significant suppression of arthritis was seen in animals receiving cyclophosphamide which was associated with a decrease in anti-collagen antibody levels. Collagen-induced arthritis in mice should be further investigated as a model to study the long term effects of "slow-acting" anti-rheumatic drugs.

Distribution of antibodies against denatured collagen in AIDS risk groups and homosexual AIDS patients suggests a link between autoimmunity and the immunopathogenesis of AIDS.

Autoimmunity often precedes the onset of AIDS-related complex or AIDS, and a number of autoantibodies have been described in AIDS patients and persons at risk for AIDS. The presence of such antibodies provokes speculation that autoimmunity is a component of AIDS pathogenesis. We report evidence of an autoantibody (anticollagen) common to all homosexual AIDS patients studied. High titer serum reactivity against collagen was detected in all homosexual AIDS patients, and in HIV+ homosexuals (66%), HIV+ i.v. drug users (38%) HIV- homosexuals (32%), HIV+ transfusion recipients (22%), and HIV+ hemophiliacs (13%), but not in HIV- i.v. drug users, HIV- transfusion recipients, HIV- hemophiliacs, rheumatoid arthritis patients, or controls. Anticollagen reactivity does not correlate with serum IgG levels, so it is not merely a reflection of polyclonal B-cell activation. Titration of anticollagen positive sera typically revealed anticollagen antibody titers 100 times those of normal sera. Affinity purification and immunoblot analysis confirmed the antibody nature of the anticollagen reactivity. The anticollagen antibodies react preferentially with primary determinants of types I and III collagen revealed after heat denaturation. Similar antibodies occur infrequently in rheumatoid arthritis patients, more often on SLE, and frequently in graft vs host disease and lepromatous leprosy. Levels of anticollagen activity in HIV+ i.v. drug users and transfusion recipients correlate with serum beta 2-microglobulin levels, suggesting that those persons with anticollagen antibodies are at greater risk of developing AIDS. This correlation, the fact that anticollagen antibodies occurred in all homosexual AIDS patients tested, and the occurrence of antibodies against denatured collagen in immune disorders with features similar to AIDS suggest these antibodies may be related to disease progression. The association of anticollagen autoantibodies with AIDS and certain other infections and immune disorders may reflect common immunopathogenic features in the etiology of these disorders.

What do anti-collagen antibodies mean?

Revue de la litterature. Relation entre le type de collagene d'un tissu et les proprietes de ce tissu. Rappel de la structure de base des collagenes. Description des pathologies dans lesquelles peuvent etre retrouvees des anticorps anti-collagene (anticollagene de type I et II essentiellement): partie centree sur la polyarthrite rhumatoide. Role et signification des anticorps anti-collagene. Association avec les groupes HLA. Etudes realisees chez l'homme et chez l'animal

A CASE OF COLONIC RUPTURE WITH EHLERS-DANLOS SYSNDROME

A case of spontaneous rupture of the sigmoid colon with Ehlers-Danlos syndrome in a 60-year-old woman is reported, whose life could be successfully saved. The patient was admitted with sever lower abdominal pain. On laparotomy a 12cm perforation was found on the antimesenteric side of the sigmoid colon, and Hartmann's operation was performed. She was diagnosed as Ehlers-Danlos syndrome by clinical signs and histopathological findings of the skin, but immunofluorescence with anti-collagen antibody showed that the distribution pattern of type I, III, IV and V collagen in the skin was normal. So, it seems that further examination of cultured skin fibroblasts from the patient is necessary. In the domestic literature, this is the first case of colonic perforation With Ehlers-Danlos syndrome, in which patient's life could be saved. For better management of this syndrome, it is important to avoid an increase in intraintestinal pressure. Moreover, primary anastomosis and subsequent closure of the colostomy should be avoided for the colonic perforation associated with this syndrome.

Transformation of BALB/3T3 cells by simian virus 40 causes a decreased synthesis of a collagen-binding heat-shock protein (hsp47).

The synthesis of a major collagen-binding heat-shock protein of molecular weight 47,000 (hsp47) was shown previously to be decreased after transformation of chick embryo fibroblasts by Rous sarcoma virus (Nagata, K., and Yamada, K. M. (1986) J. Biol. Chem. 261, 7531-7536; and Nagata, K., Saga, S., and Yamada, K. M. (1986) J. Cell Biol. 103, 223-229). In this paper, further study demonstrated that the total amount and the synthesis of hsp47 are also decreased by a factor of three in BALB/3T3 cells transformed by simian virus 40 (SV40). Higher synthesis was observed for BALB/3T3 cells in the resting state compared to those in the proliferating state. The synthesis of hsp47 in SV40-transformed cells, however, was consistently lower than that in normal cells irrespective of the cell density. Pulse label and chase experiments revealed that hsp47 was stable in the cells for at least 6 h and that there was no difference between normal and transformed BALB/3T3 cells in terms of the half-life. Decreases in the amount and the synthesis of hsp47 by transformation apparently correlate with the decreased synthesis of collagen in transformed cells. Immunoprecipitation using rat monoclonal antibody against hsp47 demonstrated the association of hsp47 with intracellular procollagen. Immunofluorescence studies using anti-hsp47 monoclonal antibody and anti-collagen antibody confirmed the co-localization of hsp47 and procollagen in both nonshocked and heat-shocked cells. Furthermore, we determined the biochemical characteristics of hsp47 of heat-shocked cells.

The plasma cell at the advancing front of the lesion in chronic periodontitis

This study analyses the ultrastructure of the plasma cell population of periodontitis-affected soft tissue close to the advancing front of interdental lesions. Biopsies from 20 patients and 3 control volunteers were examined: 5 with treated adult periodontitis (AP), 5 with untreated AP, 5 with treated juvenile or post-juvenile periodontitis (JP) and 5 with untreated JP. Plasma cell (PC) counts increased significantly (p less than 0.05) with lesion severity. They were absent from epithelium and sparse in the clinically healthy control specimens. Degenerate PC tended to be more numerous within JP tissue but differences were not significant (p greater than 0.05) when compared to AP. Intact plasma cells were never seen within JP superficial connective tissue. Russell bodies were small and few in number. The presence of degenerated plasma cells indicated normal formation and release of immunoglobulins within the tissues of AP and JP. Increased counts of degenerate PC and tissue destruction in JP suggested a correlation, possibly attributable to anti-collagen antibody secretion.

Possible formation of cutaneous amyloid from degenerative collagen fibers. Ultrastructural collagen changes and the immunoreactivity of cutaneous amyloidosis employing anti-type I, III, IV, V collagen antibodies.

The origins of primary cutaneous amyloid have been investigated and several possibilities are proposed. We investigated here several cases by electron microscopy and found in all cases characteristic filamentous changes which were identical with amyloid structures in the collagen bundles. These observations suggested a possible pathway of cutaneous amyloid formation from degenerative collagen fibers. Further studies were performed using immunoelectron microscopical and immunohistochemical methods employing a panel of anti-collagen antibodies to examine the reactivity of amyloid to them. Our results were much more suggestive of the collagen origin of this kind of amyloid.

An improved enzyme-linked immunosorbent assay of anti-collagen antibodies in human serum

An improved enzyme-linked immunosorbent assay (ELISA) for the determination of anti-collagen antibodies in human serum has been developed. The method is based on the use of serum samples diluted to 1 50 with heat-inactivated normal rabbit serum adjusted to pH 8.0 with solid Tris (0.05 M), NaCl(0.15 M) and 2 M HCl. The use of normal rabbit serum minimizes non-specific adsorption of immunoglobulin G onto the plastic surface of microtiter plate. The applicability of the method for the quantitation of anti-collagen antibodies in human serum is demonstrated with 290 specimens of sera from normal controls (194) and patients with rheumatoid arthritis (96).

Transforming growth factor beta (TGF-β) can delay and reduce lethal effects of E. Coli septic shock: [formula omitted] Stanford University, Stanford, Ca 94304, University of California at San Francisco, San Francisco, CA, University Hospital, Zurich, Switzerland and Genentech, Inc., S. San Francisco, CA 94080

Type II collagen (CII) and ovalbumin (OV) reactive T cell lines were developed from the lymph node cells of Louvain rats previously immunized with the respective antigen. Histologic sections of rat knees injected intraarticularly (ia) with 106 cells from line CII (VA) or CII (VII) were graded blindly. Significant synovitis (P < 0.000001 and 0.004, respectively) was evident compared to OV line recipients. Pannus developed within 48 hr, persisted for at least 4 to 6 weeks, was inducible with fewer than 103 CII cells, and required viable lymphocytes. The process was independent of anticollagen antibodies and was associated with the lines' capacity to elaborate a previously described 65-kDa CII-binding arthritogenic lymphokine. Line CII (VIII), which could not elicit a synovitis, did not produce detectable quantities of this factor. A line-and antigen-specific synovitis, comparable to the ia injection protocol, was also induced by intravenous administration of 2 × 107 CII (VA) cells and ia challenge with soluble collagen. These studies demonstrate that CII lines bearing the T helper phenotype can evoke a sustained synovitis and suggest that this experimental process is associated with the release of an arthritogenic lymphokine and not dependent on the formation of antibodies to CII.